Invited Feedstock Production and Utilization

Feedstock Production Jack Grushcow Jonathan Burbaum and Utilization With steadily increasing demands for renewable, scalable, non-food sources Monday, December 9, 2013 | of high quality feedstocks many 8:30am – 10:00am Corporations, Researcher and Government groups believe that Opportunities for Alternative Camelina can be THE industrial oilseed Feedstock Crops platform – delivering a truly game changing opportunity. The US DOE US Department of Energy and related has invested over $30 million in Investments in Camelina - Creating a camelina research projects, in Europe new industirial oilseed platform for the Framework 7 program has high value renewable feedstock invested 10 million euro in developing production a camelina supply chain and in Canada Moderator: Jack Grushcow, Linnaeus several initiatives over the last five Plant Sciences Inc. years have delivered over $15 million in Camelina crop development and Jonathan Burbaum, U.S. Department research. Camelina is a drought tolerant, non-food oilseed crop that of Energy has the potential to deliver increased revenues to the farmer while at the A sustainable biorefinery of Agave atrovirens in Central Mexico. From same time reducing global CO2 emissions. Camelina can be grown on traditional pulque, to novel a larger area than canola since it has bioproducts. 10 days shorting growing SergioTrejo-Estrada, Instituto requirement, uses 30% less nitrogen Politécnico Nacional and half the water. It is seeded and Evaluation of second generation harvested with the same equipment that growers are accustomed to use in biofuels production from native canola production. Because of this, the halophytes by chemical- crop has the potential to be produced characterization of Salicornia sinus- persica on millions of acres in Canada and the Ayah Alassali, Masdar Institute of northern US states. It can deliver renewable, bio-degradable feedstocks Science and Technology that can substitute for petroleum in a Guayule, An Established Industrial variety of applications including bio- Feedstock For Biorubber and lubricants, hydraulic fluids, greases, Biorefineries rigid foams and polymers; each Jeff Martin, Yulex having value well beyond bio-fuel. The panel will review the latest applied Abstracts research in this rapidly evolving field.

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Jonathan Burbaum, Program Director now ready for commercial of ARPA-E for advanced biotechnology development. Lignocellulose applications for biofuels and the transformation in bioethanol is limited production of biologically-based by the enzyme conversion cost, and chemical feedstocks is responsible for by the unusual concentration of roughly $30 million in camelina fermentation inhibitors from hydrolysis projects. He will discuss the ARPA-E and saccharification of agave biomass program goals and offer an overview by classical thermochemical methods. of key camelina projects. The value added and the unique characteristics of the new bioproducts, Sergio Trejo-Estrada make it economically feasible to scale Agave atrovirens or maguey pulquero up a balanced process of Agave is cultivated in the highlands of central biorefinery, which is contrasting to Mexico. Since prehispanic times, its other agave-based processing facilities rich sap, aguamiel, has been of tequila and mezcal. A projection of transformed into pulque, a mixed agro-ecological restoration of marginal lactic and yeast fermented beverage lands, through sustainable of proven functionality. After decades bioprocessing of Agave atrovirens is of abandonment, a new initiative of presented. Further biotechnological rural producers, government, and benchmarking efforts are needed academic institutions and agribusiness for the achievement of the full entrepreneurs, is building a pilot scale potential of this new biorefining plant for the sustainable integrated technology. transformation of Agave juices and biomass. The dry cool season provides Ayah Alassali, co-author Thomsen, M. for highly concentrated sap, useful for H. its transformation in lactic probiotic Introduction: Abu Dhabi exemplifies a beverages and mild fruity wines, coastal desert, where seawater could through fermentation. During the dry- be used for salt-tolerant crops warm and the rainy seasons, the (halophytes) cultivation. The produced complete biorefining of agave biomass halophyte biomass could be utilized in is achieved. Bioproducts such as feed, food and/or energy production, fructose sweetening syrup; prebiotic depending on its chemical fructooligosaccharides and inulin, are composition. In this study the UAE obtained from the agave juices, native halophyte Salicornia sinus- whereas the agave lignocellulose is persica was studied for its potential to transformed into bioethanol and be used as a feedstock for bioethanol fertilizer vinasses. Due to the ability of production. Fresh Salicornia sinus- Agave atrovirens to grow in eroded perica contains more than 65% of marginal lands, under harsh climate water. For such green biomass direct conditions, its cultivation is enforced fractionation and fermentation can be for soil and agro-ecological advantageous. This allows for water restoration. Several bioproducts are preservation and the ability to run at Red Text: Invited 2

lower dry matter in the fermentation 11.58% (washed) of which step. Chemical characterization and 61.12(unwashed) and 57.37% ethanol potential of the juice and (washed) was ash. Sugar analysis fibers of the fractionated Salicornia revealed relatively low concentration sinus-perica was examined in this of glucose, xylose, and arabinose in study. the juice fractions (7.15 g/L glu, 4.62 g/L xyl, and 2.488 g/L ara in juice of Methodology: Two batches of unwashed biomass and 6.62 g/L glu Salicornia sinus-persica (washed and 3.87 g/L xyl, and 1.86 g/L ara in juice unwashed) were juiced, where two of washed biomass) and not much main fractions were obtained (juice difference was observed between the and fibers). Washing of the fresh washed and unwashed biomass. The biomass aims to reduce or remove the fiber fractions contained 9.77 g/100 g nonstructural ash (salt deposits). Both DM glu, 6.97 g/100 g DM xyl, and fractions were tested for their total dry 8.44 g/100 g DM ara for the matter and ash content and sugar unwashed biomass and 10.25 g/100 g monomer composition (applying acid DM glu 7.79 g/100 g DM xyl, and 8.24 hydrolysis) as described in (Sluiter et g/100 g DM ara for the washed al., 2008a). The extent of glucan-to- biomass. This is comparable to the glucose convertibility was tested for lignocellulose content of the mature the fiber fraction, where it was (dry) plant (Cybulska et al., 2013).We subjected to Simultaneous tested S. cerevisiae on pulp fraction Saccharification and Fermentation from wet separation of Salicornia using 15 FPU/g DM of cellulases sinus-persica pretreated at low (Novozymes), with a pre-hydrolysis sereverity (121C for 30 and 60 time of 2 hours at 50 C with intensive mnutes). Both pretreatment shaking (120 rpm) prior to addition of conditions showed ethanol yields S. cerevisiae at 32C for 7 days. Final between 70-80%, and no significant ethanol yields, sugars and other inhibitory effect was observed in metabolites were analyzed by High pretreated hydrolysates, probably due Performance Liquid Chromatography to the low pre-treatment severity. as described in (Sluiter et al., 2008b). These results show that the green pulp of Salicornia sinus-persica can be

pretreated at low severity and the Results: The juice fractions were glucan fraction fermented to ethanol found to represent 67.78% of the by S. cerevisiae. unwashed batch and 74.09% of the washed batch. The wet fiber fraction Jeff Martin of the fresh-unwashed salicornia Guayule is a desert shrub native to biomass was found to contain 38.88% the southwestern US and northern DM, of which 19.18% is ash. Dry Mexico that now serves as one of the matter content of the juices were best feedstock options available for found to be 13.53% (unwashed) and biorefineries after more than 15 years Red Text: Invited 3

of crop science, plant breeding, and Customized by Chromatin, Renewable improvements in cultivation as well as by Nature agronomic practices by Yulex. Some of Moderator: John Fulcher, Chromatin the key milestones achieved to date Inc. with regards to Yulex agricultural program with guayule include 1) Giant reed (Arundo donax L.) as a consistency in phylogenetic traits sustainable energy crop for 2nd optimized for rubber production with generation ethanol in relation to its yields to achieve profitability, 2) refinery “wastes” establishment of the most advanced Enrica Bargiacchi, Consortium INSTM crop breeding program in more than 100 years, 3) crop establishment via Development of an integrated system direct seeding with high germination from cultivation of the cellulosic rates to replace previous costly energy crop Napier grass to approach of transplanting seedlings production of ethanol for biofuel grown in greenhouses, and 4) second Masahiro Samejima, The University of generation harvesting technology that Tokyo dramatically increases yields per acre and hauling efficiency to achieve Turning the Technology Flywheel: maximum highway weight limits per Moving Up the Yield Curve truckload. Steve Bobzin, Ceres, Inc.

While Yulex built its business around guayule for rubber production, this Abstracts feedstock offers the greatest potential for biorefineries to offer a wide array John Fulcher of products that come from mutually Chromatin is a vertically integrated complimentary processing sorghum feedstock provider with technologies. In addition to its core experience in biotechnology, seed, guayule rubber processing technology and feedstock growth, harvest, and platform, Yulex has been developing a delivery. Sorghum has seen wide array of supporting and resurgence in the US as a feedstock complimentary guayule processing for advanced bioethanol with the technologies that yield resins, waxes, added benefit of using the sorghum oils, cork, energy (electricity and stover and/or high biomass sorghum biofuels), green building materials, for the production of biogas. Sweet and specialty chemicals. sorghum is an attractive rotational crop with sugar cane in Brazil. In Monday, December 9, 2013 | addition, high biomass sorghum has 10:30am – 12:00pm great potential for cellulosic in Brazil and the US. The growing need for Advances in Reeds and Grasses global non-food feedstocks over the next 20 years will be met by a Red Text: Invited 4

combination of three primary sources: corn). Sorghum has a four-month energy crops; agricultural residues; growing season and in Brazil, sweet and woody biomass. By 2030 it is sorghum is being co-cultivated with estimated that 70 million acres of sugarcane to extend sugar harvests. agriculture will be need to supply the Customers are looking for sustainable demand for non-food biomass. By way customized feedstock solutions to of comparison: there are 50 million meet their needs in power, acres of sugarcane grown globally; 20 transportation fuel, and chemical million acres of canola in Canada; and markets. Biomass production must be 70 million acres of rice in China today. economically produced, harvested, By 2020 the projected demand for stored ,and transported. A biomass feedstock will be combination of purpose-grown energy approximately 600 million tons going crops along with crop residues may be to cellulosic ethanol, US heat and required to provide adequate power, EU power, chemicals and biomasss supplies year round while animal feed. Today’s markets for minimizing transportation and storage purpose-grown biomass for cellulosic costs. Through strategic alliances and the U.S. power industry are in the Chromatin can harvest, store, and $50 - $70 per ton range while the EU transport a wide variety of biomass to power market is $150 - $200 per ton. provide customers with the right Global markets for purpose-grown feedstock solution at the right time. biomass are emerging. In order to meet that demand, rapid increases in Enrica Bargiacchi, co-authors Sergio feedstock production will require: Miele & Antonio Pompeiano proven genetics; established cropping Giant reed (Adx) is the leading systems (agronomics); yield quality candidate among potential ligno- (cellulose, sugar, starch, etc.); global cellulosic feedstocks for 2nd cultivation; existing infrastructure; generation ethanol, under warm and economic viability (profit). One of temperate climates, for its high yield the key economic drivers for the of ethanol-per-hectare and low selection of biomass feedstocks will be ecological demands. This perennial C3 water. Agriculture accounts for greater grass is diffuse in natural landscapes than 90% of fresh water consumption. in Southern Europe and the Sorghum is well suited as a purpose- Mediterranean areas, pioneering a grown biomass given its water use is wide range of soils, including saline 85% less than sugarcane and 50% soils and metal contaminated soils. It less than corn. Sorghum is also can produce 20-30 t ha-1 DM in sustainable and scalable. Sorghum Southern Europe, usually with no can also be grown on marginal land irrigation and Nitrogen fertilization, with high yields and has a reduced due to its high photosynthetic rates environmental impact due to fewer and little photoinhibition. It is able to chemical inputs (for example herbicide reach early after establishment, and and pesticide use is 40-80% less than consistently maintain for +10 years, Red Text: Invited 5

high yields with favorable logistics In this presentation, recent (extended harvest period, and good achievements on the project will be bulk density). Undesired escaped summarized. Particularly, to reduce plants are easily controlled by the production cost of ethanol from glyphosate. The research activity in cellulosic biomass, year-round collaboration with Chemtex and operation of the factory by continuous Chemtex Agro, has aimed at supply of the feedstock is a key point. investigating: i) the plant ability to For this purpose, the multi- cultivating sustain environmental and soil stress, and harvesting system of Napier grass to better exploit marginal soils, and with a low input has been elaborated avoid competition with food crops; ii) in Indonesia as a model site of the possibility of recycling Adx cultivation. “wastes” from ethanol refinery to return to the soil most of the nutrients In pretreatment of our project, two- removed by the plant. In comparison step ammonia treatment system has with other potential feedstock sources, been developed to attain high such as wheat straw, potassium efficiency of enzymatic saccharification concentration in Adx lignin and ashes for both xylan and cellulose. As well from ethanol refinery makes these known, ammonia treatment can materials very interesting for soil cleave not only the ester linkages on fertilization and correction, with xylan and lignin, but also convert the increased sustainability of this crop. crystalline structure of cellulose I to cellulose III. After that, optimal Masahiro Samejima, co-authors selection and combination of enzymes Shigenori Morita, Shyu-ichi Mihashi & for saccharification of the pretreated Ei-taro Morita feedstock has been done. In addition, To develop an integrated system from recycle use of enzymes has been cultivation of cellulosic energy crop to considered to reduce the total enzyme production of ethanol in Japan and cost. To produce ethanol from xylose, Southeast Asian countries, Research a new fermentation process has been Association of Innovative Bioethanol developed by utilizing a novel yeast Technology (RAIB) has been strain selected from nature and then established in cooperation with the improved by non-GM mutation. University of Tokyo since 2009. The Finally, by integration of all these member companies of this association technologies, the research association are JX Nippon Oil & Energy Corp., aims to establish the total system to Mitsubishi Heavy Industries, Ltd., produce ethanol from cellulosic energy Toyota Motor Corp., Kajima Corp., crop with low production cost. Sapporo Engineering Ltd., and Toray Industries, Inc. The project has now Steve Bobzin, Ceres, Inc. been advancing to an integrated This presentation will examine how bench-scale test. agricultural technologies are improving the economics and Red Text: Invited 6

availability of feedstock, and their implications for scaling up biofuel BioGTL platform for the conversion of production in Brazil, the United States natural gas to fuels and chemicals. and elsewhere. Ceres is an integrated Sol Resnick, Calysta Energy seed company that combines advanced plant breeding and WRI'S CAT™ Technology for Carbon biotechnology to develop products Capture and Re-Use that can address the limitations of Karen Wawrousek, Western Research first-generation bioenergy feedstocks, Institute increase biomass productivity, reduce crop inputs and improve cultivation on Abstracts marginal land. Its development activities include sweet sorghum, Michael Schuppenhauer high-biomass sorghum, switchgrass Current progress towards meeting and miscanthus. The company's renewable advanced and cellulosic integrated technology platforms biofuel goals are falling short of include elite, proprietary breeding expectations due to technical and materials (germplasm), genetic financial hurdles. However, those mapping, biotechnology (traits) and efforts have focused on a narrow set agronomy, among others. Its largest of lignin containing feedstocks and the immediate opportunity is in Brazil, quest for liquid biofuels. Reviewing where the company is currently domestic and international large-scale working with ethanol mills to introduce yield data from alternative, novel its sweet sorghum hybrids as a low- feedstocks, using a biochemical cost, season-extending complement to pathway (anaerobic digestion) to sugarcane. gaseous advanced biofuels would on a broad base offer a vast novel pool of feedstocks and highly cost-competitive Monday, December 9, 2013 | production processes with 3-5 times 2:30pm – 4:00pm the yield per acres than pursuing liquid biofuel routes, while being GHG Gases as Feedstock: The New negative. In fact the electricity cost Renewable? would be at 3-15 c/kWh and fuel cost at $0.40 to $1.70 per GGE, while High Acreage Energy Yield of Novel offering an IRR of >20% and payback Energy Crops Offers Grid-Competitive of les than 4 years. We are presenting Novel Advanced Biofuels several cases with economic feasibility Moderator: Michael Schuppenhauer, how anaerobic digestion of non-food Farmatic Inc. crops such as corn straw, sugar beets, various cellulosic tropical grasses, Feasibility Analysis of Gas and Waste press cakes and organic wastes can Derived Fuels replace liquid fuels in energy and Julia Allen, Lux Research Inc. transportation fuel applications at Red Text: Invited 7

much lower cost than current fossil Sol Resnick fuels. These applications are Important progress has recently been particularly suitable for insular and made toward engineering a number of rural environments across the Pacific phototrophic and fermentative Rim. microorganisms for biofuels production. Several limitations, most Julia Allen notably the ever-increasing cost and First-generation biofuels dominate the linkage to oil prices of sugar alternative fuels market today, but feedstocks, currently prevent the issues like indirect land use exchange economical production of biofuels from and food vs. fuel drive demand for microbial systems. Exploiting new feedstocks. The glut of cheap methane, an inexpensive, domestically natural gas from shale as well as abundant carbon feedstock, trends toward the use of waste represents an attractive strategy biomass feedstock open up towards economically sustainable opportunities for new technologies biofuel production. Calysta Energy has that will alter the alternative fuels developed a genetic engineering landscape. Overlapping technologies platform for host organisms between gas-to-liquid (GTL) and (methanotrophs) capable of biomass-to-liquid (BTL) allow metabolizing methane to a variety of developers some degree of freedom to biofuels and biochemicals. The genetic choose the most attractive feedstock. tools, together with innovative BTL is inherently more capital- fermentation and bioprocess intensive than GTL due to the solids approaches, enable the rapid handling step. The future of BTL implementation of well-characterized hinges on advancements in pathways to utilize natural gas as a gasification and pyrolysis technology. biological feedstock instead of sugar. While BTL requires higher upfront capital, advantages include low cost of Karen Wawrousek certain feedstocks such as MSW as WRI’s patent-pending CAT™ process is well as the potential for long-term a novel biological carbon capture and feedstock agreements. At current oil re-use technology for the recycle of prices, most thermochemical GTL and carbon dioxide (CO2) from stationary BTL technologies will fail; we analyze emitters that is compatible for use the cost of 21 conversion pathways to with both small and large industrial show that gas and waste biomass CO2 sources. Currently R&D efforts processes can produce liquid fuels at have concentrated on the production $80 and $75 per barrel, respectively. of biodiesel using the CAT™ process, A further analysis on return on but CAT™ could also be used to investment reveals the cost produce other fuels and products. competitive pathways, as well as the Economical operation of the CAT™ impractical. process is achieved through unique biochemical systems utilized in the Red Text: Invited 8

process, systems for biomass and water recycle, and a system for Low-Cost Sugar from Energy Beets: A biomass residue conversion to largely Unexplored Opportunity nutrients. The microorganisms used in Steve Libsack, Betaseed WRI's CAT™ process show a rapid growth rate, indicating efficient American Process Cellulosic Sugar conversion of CO2 into biomass. Technologies “Sugar is the New However, unlike many biologically- Crude® in Asia” based carbon capture and re-use Theodora Retsina, American Process, systems, WRI’s CAT™ process is not Inc. dependent on light, which affords this process multiple advantages. Not only CO2 or Sugar to Vegetable Oil: can the CAT™ technology be directly Commercial Scale Production of Pure integrated to an existing stationary Vegetable Oil Through Plant CO2 source, it is able to operate 24 Cell Cultures hours a day, year-round. Deployment Karl Doenges, Sweetwater Energy of this technology with large, cylindrical reaction vessels reduces the Abstracts necessary land area to less than 2% of that required for equivalent Kef Kasdin biodiesel production from open pond A critical barrier to scale for the algae processes. Additionally, this production of biofuels and biobased closed system uses significantly less chemicals is access to reliable and water than that required for algal economical feedstock. Feedstock is open ponds algae or renewable crops, the largest component of end-product since evaporation is not an issue. costs and is subject to the variables Economic analysis of the process and challenges of various agricultural predicts that the CAT™ process- product markets, including sugarcane, produced biodiesel will be corn and biomass. Current economically competitive with approaches to the cultivation, harvest, petroleum-based diesel, and a storage, transport and processing of preliminary limited life cycle analysis crop-based products add substantial of the CAT™ process estimates that cost - when all that is required is CO2 emissions from industrial sources simple sugar. Proterro is the only can be reduced by greater than 80%. biofeedstock company that makes sucrose instead of extracting it from Tuesday, December 10, 2013 | crops or deconstructing cellulosic 2:00pm – 3:30pm materials. Using CO2, sunlight and water, Proterro lowers the cost of Sugars: Exploring New Sources sugar production to around five cents per pound, unleashing the economic Making sugar from CO2 value of biofuels and biobased Moderator: Kef Kasdin, Proterro chemicals for industry partners. The Red Text: Invited 9

company has developed a novel, of simple sugars for the production of scalable biosynthentic process that ethanol and many advanced integrates a patented, highly bioproducts. Energy beets will productive microorganism with a typically produce twice the ethanol per robust, modular photobioreactor made acre compared to corn, and may even from off-the-shelf materials. This compete with sugarcane. Specifically, process yields a fermentation-ready production costs are lower than for sucrose stream, rather than a mixture sugarcane, the pulp has much higher of sugars, allowing simple, low-cost value (as animal feed) than sugarcane downstream processing. Proterro will bagasse, the region where the crop present updates on it key can be grown is multiple times larger development milestones. The than the region suited to production of company has scaled up its novel sugarcane, and it is possible that photobioreactor design and conducted energy beets could supply sugar for a independent functionality tests that longer period each year than confirmed the unit’s robustness: the sugarcane. Therefore, energy beets photobioreactor is able to withstand have the ability to become a major category 1 hurricane winds. Because industrial crop for all types of of the innovative photobioreactor industrial sugars. In addition, they can design and materials used, Proterro be used as high energy, high value has also been able to validate low livestock feed as whole beets, or beet fabrication costs. Proterro is in the pulp: in the Southeast, beet pulp process of commissioning a pilot plant retails for over $700 per ton when in Florida and has completed a sold in 50-lb bags as horse feed. preliminary design, layout and Achieving the 2007 federal mandate associated cost estimate for a (Energy Independence and Security demonstration-scale plant. Act) to produce 36 billion gallons of renewable fuels each year by 2022, Steve Libsack, co-author David will depend on crops like energy Bransby beets. Being a simple sugar plant, it In North America, about 1.2 million can easily be utilized to produce acres of sugarbeets are currently industrial sugars without the use of grown for sugar production. These enzymes and the need to first convert beets supply over 50% of the US total starches to sugars. First generation sugar supply. Growing regions of feedstocks (mainly corn) for ethanol North America could be expanded to currently produce about 14 billion include almost every state in the U.S, gallons of ethanol each year. To and energy beets can be grown as a achieve the 36 billion gallon mandate winter crop in most southern regions. we will need about 21 billion gallons The potential use of beets for produced from second generation purposes beyond sugar for human advanced biofuel feedstocks. Energy consumption is virtually untapped. beets are an advanced biofuel Energy Beets are an excellent source feedstock, and may well qualify as a Red Text: Invited 10

cellulosic feedstock. We have recently The Green Power+® technology is a conducted energy beet trials in the patented technology for the Southeast with good success. For production of low-cost cellulosic example, an initial commercial-scale sugars from the hemicelluloses of test in Baldwin County, South biomass. Alabama, yielded 35 tons of beets per acre over the winter with no irrigation Our AVAP® technology is a patented (http://blog.al.com/live/2013/05/man technology for the production of low- ufacturing_plant_cited_as_m.html). cost cellulosic sugars from the This translates into approximately 830 cellulose and hemicelluloses of any gallons of ethanol per acre, and at a biomass. price of $30 per ton for beets, a gross income for the grower of $1,050 per The two technologies address different acre and a sugar cost of 10 - 15 cents market opportunities for biorefineries per pound for the processor. Work on in the Asia Pacific region and irrigated beet production for worldwide. For each technology, any conversion to ethanol is also underway biomass may be utilized, including in California hardwoods, softwoods, and (http://www.ethanolproducer.com/arti agricultural residues. The Green cles/9978/lowering-ethanols-carbon- Power+ process produces low-cost C5 footprint-with-energy-beets). Our and C6 sugars from the hemicelluloses panel will provide an overview of the of biomass feedstocks. Sugars are potential of energy beets for extracted from the solids which are production of biofuels and then utilized for existing applications, bioproducts, including the agronomy in synergy with biomass-based of the crop and possible end-uses. renewable power, pellets, sugarcane plants, and many other existing sites. Theodora Retsina Co-location minimizes capital costs for At American Process, we believe that commercial implementation. We see Sugar is the New Crude®. Our vision tremendous opportunity for Green is to be the leading biomass pre- Power+ configurations Asian sugar treatment technology in the world, cane ethanol facilities, where our with a value proposition that combines technology can substantially increase engineering capabilities, quality of the production of ethanol in a very technical support, assurance of high capital efficient manner. Other yields, clean sugars, and minimum applications include pellet-making total cost of operation. We have facilities and biomass power plants. developed two proprietary biorefinery Our first large-scale implementation of technologies for producing low-cost Green Power+ technology is a cellulosic sugars from non-food based biorefinery pilot plant in Alpena, biomass. Michigan, capable of converting about 20 tons/day of hemicelluloses to sugars and co-products. The plant Red Text: Invited 11

capacity is up to 2 million gallon/year biorefinery started up in the first half of ethanol. The Alpena biorefinery of 2013. started up in Q3/2012. The AVAP process produces low-cost C5 and C6 Karl Doenges sugars from both cellulose and The recent volatility of commodity hemicellulose of biomass feedstocks. prices and rapid pace of industry The AVAP process employs a solvent development is stimulating the for lignin and sulfur dioxide to cleanly biorefining and oleochemical industry fractionate biomass into cellulose, to diversify its raw material feedstock, hemicellulose, and lignin. Optionally, using an array of fats and oils. The the cellulose may be recovered as a ability to take various fats and oils is a cellulose material or precursor for key risk mitigation strategy; however, making cellulose derivatives. The reengineering the way oils are made cellulose that is produced by the AVAP addresses the feedstock problem at a process is extremely susceptible to more fundamental and sustainable hydrolysis by enzymes to produce level. This presentation will review glucose, thus greatly reducing enzyme Sweetwater Energy’s deployment of a costs. We see AVAP as having new CO2 or sugar to pure vegetable tremendous potential in Asia in areas oil technology, whereby a modular, of abundant, competitively priced CO2 source-flexible, commercial scale biomass feedstock. An advantage of oil production technology has been this technology is that it can operate optimized for the biodiesel, renewable with softwood, hardwood or diesel and oleochemical agricultural residue, thus increasing industries. This talk will highlight: 1) the number of locations where it can The basic science behind CO2 and be deployed. The cellulose sugars sugars to pure vegetable oils, 2) using from the AVAP process can be this technology as a bolt-on for corn converted into a variety of high-value ethanol plants and the resulting chemicals, many of which are high synergies, and 3) characterization of demand in Asia either on their own or the vegetable oil to enable optimum as a feedstock for plastics downstream processing manufacture. Our first large-scale implementation of AVAP technology is Tuesday, December 10, 2013 | a biorefinery pilot plant located in 4:00pm – 5:30pm Thomaston, Georgia. The Thomaston biorefinery is capable of converting Renewable Oil Feedstocks for about 10 tons/day of biomass to the Pacific Rim sugars, ethanol, and co-products. The plant capacity is up to 300,000 Moderator: Sanjay Wagle, AliphaJet gallon/year of ethanol or other products, such as butanol, jet fuel, or Naveen Sikka, TerViva biochemicals. The Thomaston

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Mike Cey, Ag-West Bio Inc Oilseed Derived Feedstocks - Can they Chris Tindal, Office of the Deputy compete with Petroleum? Assistant Secretary of the Navy for

Energy The conversation related to oilseed Abstract derived feedstocks has been Speakers will discuss how they have traditionally dominated by bio-fuel begun to commercially establish their applications. As the reality of the feedstocks, with specific emphasis on: disparity between production costs (1) Business models -- contemplated and the market price of oilseed structures, process for validation, and sourced bio-fuels sets in, attention is ultimate winning models with customers (2) Prioritization -- how the shifting to higher value oilseed feedstock company prioritized applications such a polymers, landowner customer segments, lubricants, surfactants, cosmetics and geographies, and downstream market speciality chemicals. Addressing these opportunities (3) Focus on Asia Pac -- markets requires a combination of evaluation of feedstock advanced oilseed profiles and appropriateness for Asia Pacific processing methods working together countries. to develop next generation bio- Wednesday, December 11, refineries. The assembled panel will 2013 | 8:30am – 10:00am discuss four unique approaches to delivering high value renewable Oilseed Derived Feedstocks - products from oilseed feedstocks with Can they compete with value beyond bio-fuels. Petroleum?

Moderator: Jack Grushcow, Linnaeus Plant Sciences Inc. Renewable Chemical

Jonathan Burbaum, U.S. Department Platforms of Energy Monday, December 9, 2013 | Andy Shafer, Elevance Renewable 8:30am – 10:00am Sciences, Inc. Differentiation for Bio-based David C. Bressler, Biorefining Chemicals Conversions Network

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Moderator: Damien Perriman, Elevance is now working with its Genomatica partners such as US-based surfactants producer Stepan to meet product Jeffrey Hsu, Far Eastern New Century demand and accelerate deployment and commercialization of their high- Commercialization - Elevance's World performance chemicals in end user Scale Biorefinery has commenced applications. This paper will discuss production and New Products have the status and progress of the entered the Market - An update on commercialization activity taking place progress and what's coming and the benefits the bio-based Andy Shafer, Elevance Renewable technology is bringing to the Sciences, Inc. downstream markets.

Rodolfo Manzone, Yulex Rodolfo Manzone Yulex applies crop science, Abstracts bioprocessing and materials science for Damien Perriman the development of non-GMO, plant- Jeffrey Hsu based biomaterials from guayule, a In addition to all the other challenges sustainable and industrial crop. of commercializing bio-based Guayule is a highly productive chemicals, Asian markets are “natural” evaluating the potential impact of chemical factory which requires low ‘low-cost’ petrochemical plants now inputs without competing with fiber being built in China. This discussion and food crops. will examine how multiple companies The company is developing fully are looking to highlight the value of integrated and sustainable their bio-based offerings, to have biorefineries designed to utilize successful market entry and growth virtually 100% of guayule to provide despite the low-cost challenge. a renewable source of rubber emulsion, solid rubber, resins Andy Shafer and biomass on a global basis. Yulex's Elevance Renewable Sciences and biomaterials are tailored to Wilmar International Limited have replace traditional tropical Hevea or begun commercial shipment of petroleum based rubber for specialty chemicals from their World- consumer, industrial and medical Scale biochemical refinery in Asia. The markets, with the residual biorefinery, which has a capacity of agricultural materials utilized as a 180 KTPY (400m lbs.) and is feedstock for bioenergy. expandable to 360 ktpy, is Elevance's first biorefinery using its proprietary Yulex’s technological innovation spans metathesis technology. With from guayule seed technology, commercial capacity now available, Red Text: Invited 14

agronomics, breeding, harvesting, molecules in the current oleochemical bioprocessing to materials value chain such as fatty acids, fatty science. Yulex develops natural alcohols, and dimer acids that are rubber both in liquid and solid form. used in the manufacture of consumer Resin can be produced both as a products ranging from surfactants and crude extract as well as a lubricants to polymers and cosmetics. fractionated refined resin These oleochemicals provide a component. Leftover biomass can renewable alternative to either be petrochemical-based products. New used for biomass energy to produce technologies are being developed that electricity in its raw form, or it broaden the reach of oleochemicals can be processed to produce both into new products and are providing solid and liquid biofuels. bio-based alternatives for consumers. Significant progress has been made This panel assembles speakers with regards to specialty chemicals representing multiple points along the as well. value chain of converting oleochemical inputs into bio-based consumer Yulex is based in the U.S. Southwest products. From plantation and and has a growing global footprint. vegetable oil refining to renewable chemical conversion technology to Monday, December 9, 2013 | polymerization and product 10:30am – 12:00pm applications, the presentations show the current value chain as well as the Expanding the Oleochemical future potential value chain of the Value Chain to Bio-based oleochemical industry.

Consumer Products Allen Barbieri Moderator: Tom Beardslee, Verdezyne OVERVIEW

Biosynthetic Technologies (BT) holds Hirzun Mohd Yusof, Sime Darby exclusive patented technology that

synthesizes plant and animal oils into Sam Bhargava, Jarden Applied high performance synthetic oils used Materials in lubricants, industrial chemicals and

personal care. These “biosynthetic” Biobased Synthetic Chemistry base oils meet or exceed the Allen Barbieri, Biosynthetic performance characteristics of existing Technologies petroleum based synthetic oils. Having

tested BTs biosynthetic oils Abstracts extensively, several of the world’s Tom Beardslee largest manufacturers of automotive Hirzun Mohd Yusof and industrial lubricants are certifying Sam Bhargava finished products they will bring to The conventional processing of crude market under their brand names. The vegetable oils produces an array of Red Text: Invited 15

chief formulator of a leading global exceed the industry standard of 60% motor oil company recently stated biodegradation within 28 days. that these are “the most exciting products to enter the lubricants Non-Toxic: BTs oils tested non-toxic, industry since the introduction of PAOs even at contamination levels 100 [synthetic lubricants] 50 years ago.” times higher than the level needed to First-mover advantage, lack of viable claim non-toxicity. competition and patented technology make BT a market leader in the Not bioaccumulative: BT’s products do sustainable chemicals sector. not bio-accumulate in living organisms. Business model: BT is a wholesale manufacturer/distributor of biobased Greenhouse gas reduction: A Life synthetic oils that can be used by Cycle Analysis of Greenhouse Gas finished motor oil, lubricant and Emissions (GHG) from BT’s products is chemicals products manufacturers. 83% lower than the GHG emissions BT is currently working with dozens of associated with poly-alpha olefin leading global lubricant and additive (PAO), a product of similar function manufacturers as well as automotive and use. and equipment manufacturers who are each testing, formulating and Machine life / fuel efficiency: preparing to launch motor oils and Numerous tests have shown that BTs lubricant products based on BT’s biosynthetic oils keep metal surfaces biosynthetic oils. cleaner and facilitate lower friction and scarring on bearing surfaces than the Manufacturing: BT contracted with highest quality synthetic oils. All of Albemarle to build a demo this keeps leads to longer machine manufacturing plant which is currently life, lower maintenance costs and operating in Baton Rouge. In late fall, increased fuel efficiency. 2013, BT will begin construction of a full scale continuous flow production Water pollution: BTs biodegradable plant. oils could significantly reduce the environmental impact of improperly ENVIRONMENTAL BENEFITS dumped motor oil and oil runoff from roads, which accounts for 40% of Sustainable: BTs oils are made from water pollution in the U.S. (EPA). In fatty acids found in plant oils including fact the amount of used motor oil and canola, soy, palm, and coconut, as lubricants that enter the world’s well as refined animal fat (all 100% oceans is the equivalent of one Exxon renewable carbon). Valdez spills every week.

Biodegradable: Using the most Monday, December 9, 2013 | stringent testing protocols, BTs oils all 2:30pm– 4:00pm Red Text: Invited 16

Metabolix is also focused on Renewable Chemicals and developing biobased four-carbon (C4) Consumer Products and three-carbon (C3) chemicals. The C4 program is currently at the Renewable Chemicals: The Path to semiworks scale (80,000L) and Commercialization Metabolix has shipped samples of Moderator: Max Senechal, Metabolix biobased gamma butyrolactone (GBL) to prospective customers for testing, DEINOL: The Deino Way to Biofuels. and GBL has been successfully Emmanuel Petiot, Deinove SA converted to BDO via hydrogenation. Metabolix is making steady progress Leveraging a Consumer Brand for B2B with its C3 program as well – the Pull - The Nexa Project Company is currently running Jeff Uhrig, Bioformix fermentation at the 20L scale and recently successfully recovered acrylic Chemo-catalytic conversion of acid from biomass using its cellulose into para-xylene proprietary FAST process. In 2013, John Bissell, Micromidas Metabolix is continuing fermentation scale up, engineering of microbial Abstracts strains, and development and optimization of its FAST recovery Max Senechal technology to produce biobased C4 Metabolix, Inc. is an advanced and C3 chemicals to match chemical biomaterials company that is well industry specifications for quality and positioned to address growing market purity. demand for sustainable solutions in the plastics, chemicals and energy In this presentation, Metabolix’s vice industries. The Metabolix scientific president, biobased chemicals, Max foundation and core science is the Senechal, will outline the Company’s metabolic pathways for the production strategy for bringing these renewable of a class of microbial biopolymers – chemicals to market and review polyhydroxyalkanoates (PHAs) – from various strategic options available to renewable resources. PHAs can be biobased chemical producers in tailored through metabolic pathway commercializing their innovative engineering and have a wide range of products. Key question addressed: Is applications in industry, including use the “drop-in” chemicals strategy in polymer form as biobased plastics currently pursued by several or in monomer form as chemical participants in the space the only intermediates. The first commercial approach worth considering for product using the fermentation route successful market entry? are biopolymers marketed under the Mirel and Mvera brands. Emmanuel Petiot

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DEINOVE (Alternext Paris: ALDEI) is a generally concentrated on feedstock, cleantech company that designs, enzymes and capital, with feedstock develops and markets novel industrial and biotransformation accounting processes using biotransforming generally at least 50% of the total platforms based on the untapped cost. However, most processes are extraordinary and advantageous constrained to operate at the optimal properties of the Deinococci. Originally temperatures of yeast or commonly founded in France, and principally used bacteria (~35C), not maximizing located in Montpellier, DEINOVE is on biotransformation kinetics, favoring now offering its technology platforms infections and preventing from using globally, with commercial enzymes at their optimum activity representation in both the EU and levels. United States (www.deinove.com). DEINOL’s platform leverage Taking advantage of the unique thermophillic organisms (45-55C) that genetic properties and unusual are not subject to these constraints or robustness of this relatively unknown limitations, and release their own bacterial phylum, DEINOVE optimizes enzymes in solution during the natural fermentation and metabolic fermentation/hydrolysis, a rare capabilities of these bacterial "micro- process advantage named factories" to produce renewable green Consolidated Bioprocessing (CBP). chemicals. The company is structured Having the ability of produce end- in two groups: while DEINOL is products and expressing the enzymes focused on producing renewable necessary to break down feedstock for advanced biofuels, DEINOCHEM is metabolism, all at higher focused on green chemicals or temperatures, gives DEINOL’s intermediates. Of notice, two solutions a unique and significant cost subsidiaries operate independently, advantage in techno-economic yet falling within the DEINOCHEM analyses. group: new antibiotics research (DEINOBIOTICS SAS) and plastics Of particular notice, DEINOL’s bioremediation (THANAPLASTTM). organisms co-assimilate C5 and C6 sugars as well as glycerol and acetic DEINOL is focused on producing acid, and are particularly resistant to bioalcohols and alkenes of interest for process inhibitors, thereby maximizing their use as high octane (RON) drop-in both conversion rates and feedstock components of transportation fuels. utilization. When considering their The industrial production of biofuels, incredible advantages including through a biochemical route, feedstock flexibility or their efficient necessitates the hydrolysis of a metabolism of all organics, Deinove’s feedstock, and its fermentation to strains truly are the ideal hosts for generate the end product of interest. biotransforming applications by The main costs of production are allowing for maximized process Red Text: Invited 18

efficiencies and reduced raw material other aromatic species. The and variable costs. presentation will discuss (1) the specific advantages of the chemical In summary, the exceptional pathway, (2) which feedstocks have properties (robustness, biodiversity been tested and will be scaled first, and metabolic properties) of DEINOL’s (3) the comparative projected platforms, and DEINOL's economics of the process, and (4) the developmental research results, performance of our recently demonstrate the huge potential for constructed pilot plant. these microorganism’s to become true bacterial micro-factories for the Tuesday, December 10, 2013 | production of biofuels in a cost- 2:00pm– 3:30pm effective, environmentally friendly way. Processes Leading to Commercialization Jeff Uhrig

While B2B sales channels are Making it Real: Levulinic Acid as a attractive to resource limited start- Biobased Building Block of a ups, the consumer channel presents a Renewable Chemical Industry nearer term commercial opportunity, Moderator: Atul Thakrar, Segetis allowing companies to build brand equity, demonstrate market adoption Strategic Commercialization of Bio- and de-risk larger scale B2B based Chemicals applications. This presentation will Sue Hager, Myriant Corporation address how Bioformix created a consumer brand - Nexabond 2500 - Developing new platform molecules: a and a respective in internet and case study consumer distribution channel in 12 Marcel Lubben, DSM months, which has in turn created stronger pull from global Bio-butadiene: Techno-economics of manufacturing partners. bioprocessing route via 2,3 butanediol

(BDO) John Bissell Jayant Sawant, Praj Matrix – The Micromidas has developed a Innovation Center chemocatalytic route to para-xylene from cellulosic biomass and ethylene. Abstracts The process is expected to be cost- advantaged over naphtha-derived Atul Thakrar para-xylene, and will enable the Segetis creates high performance, production of 100% bio-based PET. sustainable materials, reducing the The highly selective synthesis allows world's dependence on fossil fuel for high yields of para-xylene without based petrochemicals. Our Levulinic the presence of typical isomers or acid based technology platform, Red Text: Invited 19

including the ketal-based JAVELINTM performance, healthier, biobased Technology, is positioned for materials. significant global adoption with renewable alternatives that cut Sue Hager greenhouse gas emissions and The chemicals industry is experiencing address toxicity issues in everyday a fundamental shift as cost- items. Segetis has successfully competitive bio-based chemicals developed and commercialized become a commercial reality. Leading specialty plasticizers for flexible PVC this transformation is the market used in everyday objects including availability of bio-succinic acid, a flooring, shoes and packaging, as well platform molecule used as a direct as formulation aids used in consumer replacement for petroleum-based and industrial markets such as succinic acid and also as a detergents and hard surface cleaners. fundamental building block chemical in the production of numerous industrial Levulinic Acid, the starting point for and consumer applications. Myriant is these innovations, is the one of the world leaders decomposition product of sugars and commercializing bio-succinic acid with carbohydrates, the most abundant two operating plants in production. feedstocks on Earth. Levulinic Acid is Myriant’s presentation will discuss the readily transformed into a wide variety company’s pathway for strategic of chemicals and has the potential to financing of its flagship commercial become a foundational biobased plant, its commercialization strategy chemical building block of a renewable for the production of bio-succinic acid, chemical industry. Despite high levels as well as its plans for accelerated of interest, the lack of large-scale, development of its bio-based cost effective, clean Levulinic Acid chemicals pipeline. production has hampered the growth of levulinic-based products. Segetis Marcel Lubben has made a breakthrough in making Royal DSM N.V. is a global science- Levulinic Acid a reality with a based company active in health, disruptive process that enables nutrition and materials. DSM is a industrial-scale Levulinic Acid frontrunner in creating bio-based and production using a wide array of environmentally sound solutions, and biomass feedstocks, including seeks to demonstrate the commercial fructose, glucose and cellulosic sugars. viability of renewable technologies in The combination of our ability to make collaboration with strategic partners in cost effective Levulinic Acid at scale the value chain. The development and with the market development for supply of high value knowledge, JavelinTM plasticizers and formulation ingredients and expertise in the field aids positions Segetis to meet the of bio-conversion technology are increasing demand for high critical success factors determining its future. DSM’s strategy is to license its Red Text: Invited 20

technology and expertise to bio-based Butadiene (BD) is a top 10 entrepreneurs, enabling them to petrochemical primarily produced by convert biomass in the most heavy crackers. With more US commercial and sustainable ways crackers shifting to shale gas based possible. Today’s market needs are light crackers, a net drop in production driven by a number of major global of butadiene has created a demand- trends and challenges. At DSM we are supply imbalance and sharp price using our innovative strengths to swings in global markets. address some of the most important Alternatively, butadiene can be of these trends and challenges focused produced from renewable feedstocks on supporting the transition from such as sugars using greener routes fossil to renewable raw materials. The via 2,3 butanediol-fermentation and investment wave in the bio-based subsequent conversion to BD by economy started from the early 00s. catalysis. Several research groups, After almost one decade, tremendous start-ups and few petrochemical and impressive progress has been majors have shifted focus to bio- made in technology development for butadiene. The focus of our study is to renewable chemicals and materials. develop deeper insights into the Pioneers in the field are making their technical and market driven first commercial steps. challenges of bio-BD production and shortlist solutions to overcome the In this speech we shall present a case challenges. We present a detailed study to illustrate our vision on what techno-economic analysis of bio- drives success in the butadiene production starting with commercialization of bio-based sugar based BDO fermentation and chemicals and materials: partnerships purification by solvent extraction & capital, feedstock choice, excellent followed by catalytic conversion of operations, application know-how and BDO to BD. Detailed sensitivity value chain understanding. Building on analysis and case scenarios with the experience DSM has assembled different sugar sources, sugar prices, over the past years in bio-conversion and process parameters, and break- technologies and the creation of even analysis for crude versus sugar chemical building blocks for based BD are presented. (performance) materials from bio- based feedstock, an appeal is made to Tuesday, December 10, 2013 | view the bio-based economy as an 4:00pm – 5:30pm opportunity to innovate ourselves out of the global economic crisis. Novel Fine Chemicals Manufacturing and creative approaches, Using Biocatalysts collaborations and partnerships are needed and will be discussed. Biocatalytic Preparation of

Cyclopropanes – A Novel and Cost- Jayant Sawant Effective Approach Red Text: Invited 21

Moderator: Pedro Coelho, Provivi, Inc Our initial product platform is based on breakthrough technology for Satish Khanna, Provivi,Inc cyclopropane biosynthesis developed Ajay Parikh, ZCL Chemicals at and licensed from the California Anand Ghosalkar, Praj Corporation Institute of Technology. There, we developed the most active catalyst ever reported for olefin Abstracts cyclopropanation, a key reaction in the synthesis of drugs, hormones, and Satish Khanna insecticides (Coelho et al., Science Ajay Parikh 339, 307 (2013); Coelho, et al. Nat. Anand Ghosalkar Chem. Biol. 9, 485 (2013)). The In recent years, biocatalysis has combined annual global market size established itself as a key technology for products that are accessible via for the production of fine chemicals. Provivi’s technology exceeds $7 The fine chemicals industry utilizes billion. Active pharmaceutical this technology to catalyze chemical ingredients (APIs) that contain a reactions with unprecedented regio cyclopropane motif have a combined and enantioselectivity. market of over $5 billion. In addition, Implementation of biocatalysis pyrethroid insecticides containing a therefore often results in a decreased cyclopropane motif have a market size number of chemical steps required for of about $2 billion. Here we will briefly the synthesis of a given fine chemical, describe the technology, and then and ultimately reduces the cost of focus on product applications and manufacture. In addition, biocatalysis market opportunities pursued by favors mild conditions, resulting in Provivi. decreased waste streams, decreased energy input, and improved Wednesday, December 11, safety. With an ever expanding 2013 | 8:30am – 10:00am biocatalysis toolbox and decreasing enzyme engineering timelines, Innovation in Renewable biocatalysis is being embraced by the Chemical Platforms fine chemicals industry today. This session will discuss the value Bio-based Innovations and New biocatalysis provides to a number of Ventures from the Engineering prominent fine chemical Research Center for Biorenewable manufacturers, as well as challenges Chemicals (CBiRC) and opportunities. Moderator: Peter Keeling, Iowa State

University,OmegaChea and SusTerea Pedro Coelho

Provivi, Inc. is a start-up company Karl Sanford, DuPont/Danisco aimed at producing high-value chemicals via biocatalytic processes. Joe Noel, Pareto Biotechnologies Red Text: Invited 22

Jeff Fornero, Glucan Biorenewables Moderator: Brian Conroy, BP

Abstract The Reality of Commercial Scale Cellulosic Ethanol Production CBiRC is developing renewable Christopher Standlee, Abengoa chemical platforms from bio-based Bioenergy US Holding, Inc. innovations and new ventures utilizing tools, components and materials being Meeting worldwide demand with explored by the Center. Core cellulose knowhow and technologies include Steve Hartig, POET-DSM Advanced bioengineering of fatty acid and Biofuels polyketide biochemistry in microorganisms, as well as an Delivering Big by Getting Small innovative and complimentary Wes Bolsen, Cool Planet Biofuels portfolio of developments in chemical catalysis. By combining biocatalysis Abstracts and chemical catalysis CBiRC creates new knowhow and powerful Christopher Standlee technologies that have the potential to Abengoa is an international leader in nurture a sustainable bio-based the development of Advanced chemical industry. CBiRC believes the Biofuels. We have pursued a 10 year existing petrochemical supply chain effort to produce cellulosic ethanol can be transformed with key from multiple agricultural residues, foundational intermediates that deliver culminating in the commissioning and an array of drop-in chemistry or start up of our Hugoton, Kansas similar functionality to existing fossil- commercial scale cellulosic facility in carbon-based chemicals. Here we will December 2013. Abengoa also started describe our progress towards operations at a demonstration scale creating an advanced manufacturing municipal solid waste to ethanol system with new platform molecules facility in Spain, and is negotiating a for biobased chemicals. commercial scale MSW facility today. We are also pursuing efforts to Advanced Biofuels and produce other advanced biofuels and bioproducts from the same agricultural Biorefinery Platforms residue and MSW feedstocks. We currently are sourcing and managing Monday, December 9, 2013 | 380,000 tons of ag residues annually for our Kansas facility, and have 8:30am – 10:00am already conducted 4 harvests. We also

are planning to utilize dedicated Cellulosic Biofuels: Unleashing energy crops at some point in the near Commercial Production future. These efforts, along with start Red Text: Invited 23

up of the Ineos facility and progress in Biofuels is constructing a 25 million- development and construction by gallon-per-year cellulosic bio-ethanol other producers of Advanced Biofuels plant in Emmetsburg, Iowa that will make second generation biofuels a use corn cobs, leaves, husk and some reality. stalk as feedstock. Farmers this fall will harvest an estimated 120 Steve Hartig thousand tons of biomass to feed this Cellulosic bio-ethanol is poised to play commercial biorefinery. Construction a crucial role in the energy landscape is on schedule for startup in early of the United States, driven by recent 2014. POET-DSM is now working to process and technology gains as well license this technology to other as a commitment by government ethanol producers, and we see Asia as leaders to prioritize domestic, an important opportunity for putting sustainable energy production. But this technology to good use. Sound worldwide, the potential is even technology, created here in the U.S., greater. In the U.S., liquid fuels is ready to provide solutions for demand has fallen in recent years, energy demands well into the future. and most experts predict this trend will continue. However, worldwide Wes Bolsen demand, driven largely by growth in Carbon negative fuels is a reality Asia, is expected to increase today through the combination of dramatically in coming decades. The biofuels and biochar. Wes will talk potential for expansion of cellulosic about the company’s ability to bio-ethanol in those areas is commercialize Cool Planet’s enormous. China, as the second- technology through a much smaller, largest corn producer in the world, is distributed plant model. He will lay uniquely suited to reap the rewards of out transformative industry models commercialization efforts such as such as mainframe computers those by POET-DSM Advanced becoming personal computers, and Biofuels. China has stated that it will the mass production of micro-refinery not use corn for ethanol production; technology paralleling other however, the abundance of crop industries. He will finish by discussing residue that remains after harvest their first commercial facility and how does provide ample feedstock for with the backing of Google, GE, biomass-based ethanol production. ConocoPhillips, BP, and their other The Energy Information Administration marquee investors, they were able to expects China to lead the world in commercialize. demand growth for liquid fuel by 2035. India is expected to experience Monday, December 9| 10:30am the next-largest demand growth, and – 12:00pm the rest of Asia is not far behind. Cellulosic bio-ethanol can help meet that demand. POET-DSM Advanced Red Text: Invited 24

Production of Drop-In particularly desirable for heavy-duty Hydrocarbon Fuels from vehicles and air travel. Furthermore, Cellulosic Biomass enzyme costs for biological conversion of lower cost cellulosic biomass to Moderator: Thomas Foust, National ethanol tend to be high. Catalytic Renewable Energy Laboratory routes are being developed to convert biomass sugars and their breakdown Charles Cai, University of California, products such as furfural, 5- Riverside hydroxymethylfurfural (HMF), and levulinic acid that we call fuel Jesse Q. Bond, Syracuse University precursors into hydrocarbon fuels and fuel components compatible with the DME, its advantages and its promise existing fuel infrastructure. A potential through small-scale production advantage of this aqueous processing Brittany Syz, Oberon Fuels approach to such “drop-in” fuels is that only thermochemical conversion Abstracts is needed to make fuel precursors from cellulosic biomass, thereby Thomas Foust avoiding use of expensive enzymes. Charles Cai However, yields for traditional Jesse Q. Bond approaches to producing such Among sustainable resources, biomass intermediates are too low to be cost- is uniquely suited to production of competitive, and the key economic organic fuels on which our society is challenge is developing low cost so dependent, and only cellulosic routes that realize high yields of fuel biomass offers low enough costs and precursors from cellulosic biomass sufficient abundance to make a large that can be integrated with catalytic impact on petroleum use. However, conversion to hydrocarbons. although biomass at about $60/dry ton has an equivalent energy cost to This panel will provide insights and oil at about $20/barrel, conversion of recent performance information for cellulosic biomass is currently the key operations, leading expensive, and new processes are technologies, and commercial needed that overcome recalcitrance as strategies for aqueous processing of the primary barrier to cellulosic biomass for catalytic competitiveness. In addition, ethanol conversion to fuels. Charles Cai from and other oxygenates made from the University of California Riverside biomass carbohydrates, currently will start with an overview of biomass mostly sugar and starch and latter deconstruction to form fuel precursors cellulosic biomass, are not fully including sugars, furfural, HMF, and compatible with the current fuel levulinic acid. Various routes to infrastructure. And hydrocarbon fuels produce these components will be with higher energy density are described, yield and operational Red Text: Invited 25

considerations will be discussed, and alternative that will meet strict perspectives will be offered on leading emissions standards. options and challenges. DME has been used for decades as an Next, Jesse Bond from Syracuse energy source in China, Japan, Korea, University will summarize pathways Egypt, and Brazil, and it can be for catalytic processing of these fuel produced domestically from a variety precursors and identify appropriate of feedstocks, including biogas (plant catalysts for each fuel option. In and food scraps) and natural gas. addition, theoretical yield limits and Ideal uses in North America are in the current performance will be described transportation, agriculture, and for the various fuel options, and construction industries. Because opportunities for improvements will be production is not dependent upon the outlined. Tom Foust from the National price of crude oil, DME will have stable Renewable Energy Laboratory (NREL) pricing, which is competitive with that will then provide an overview of the of diesel. National Advanced Biofuels Consortium (NABC) and describe DME has storage and handling NABC partnerships to accelerate advantages over other diesel commercial production of hydrocarbon alternatives, such as CNG and LNG. In fuels. Included will be an outline of current demonstrations using bioDME key metrics that must be met for in Europe, Volvo has shown 95% commercial viability and the stage of reduction in CO2 emissions with development of various processes to companies such as DHL using DME in meet these goals. commercial operations. See http://www.biodme.eu The range of topics covered in this panel will allow the audience to better Oberon Fuels has developed an understand the pathways, progress, innovative, small-scale production and challenges in converting cellulosic method that produces DME in regional biomass into fuels and fuel fuel markets, bypassing the initial components that are compatible with need for a national infrastructure. In the existing aircraft and heavy and these regional fuel markets, regional light duty vehicle infrastructures. feedstocks are used to produce DME to be utilized in regional trucking Brittany Syz hauls, creating new regional DME (dimethyl ether) is a clean- economies and green jobs. The first burning, non-toxic, diesel alternative. such production facility will be online Its high cetane value and quiet in Imperial Valley, in Brawley, CA in combustion, as well as its inexpensive August 2013. The regional model propane-like fueling system, make it supports not only local job growth and an excellent, inexpensive diesel clean air initiatives, it also provides a viable economic structure that allows Red Text: Invited 26

for capital expenditure in sync with performance level of any of the diesel market growth – it really hits the alternatives. triple bottom line concept of sustainability. On June 20th, Mack Trucks also announced they plan to commercialize The need: DME trucks. In its press release, Mack Trucks noted that DME “offers many In his climate initiatives speech in environmental and societal benefits, July, President Obama identified that including that it can be made from we need to address heavy-duty multiple sustainable feedstocks,” trucking. Why? Because 30% of the including grass clippings, animal waste particulate matter in our atmosphere and other sustainable sources. is caused by commercial trucking. DME produced using the Oberon Fuels See:http://www.aboutdme.org/aboutd method is actually carbon negative – me/files/ccLibraryFiles/Filename/0000 because it uses feedstocks that would 00002405/Release_Mack_2013-06- otherwise release methane into the 20.pdf atmosphere and converts them to a clean-burning, zero particulate matter Conclusion: fuel. DME is the smart choice for an Partners entering the field: alternative fuel to diesel. Our goal is to educate your audience on the In early June, on the steps of the prospects and promise of DME and the Capitol in Sacramento with small-scale production method that representatives of the Governor’s models its success. office involved, Oberon Fuels, Volvo Trucks North America and Safeway Monday, December 9| 2:30pm Inc. (the big CA-based food giant) – 4:00pm announced a partnership to demonstrate DME in the San Joaquin Biorefineries: Perspectives on Valley. They have received a Finance & Construction $500,000 grant from the San Joaquin

Valley Air Pollution Control District to Moderator: Martin Sabarsky, Cellana help support the project. LLC

At the same event, Volvo Trucks North US next-generation biofuel America announced plans that it would investment: no swing for the fences commercialize DME trucks in North yet America by 2015. Volvo is the largest Alejandro Zamorano Cadavid, seller of heavy-duty trucks worldwide. Bloomberg New Energy Finance It is committed to DME engines because DME has the best Red Text: Invited 27

Decision and Risk Analysis in business case to attract needed Industrial Biotechnology funding. This presentation will Alan Propp, Merrick & Company describe the Decision and Risk Analysis process and present an Project Financing Considerations example of how the science was Lester H. Krone, Stern Brothers & Co. applied to an industrial biotechnology project. Critical Questions for Bolt-On Biorefineries: Avoiding Unintended Lester H. Krone Consequences Project Finance Fundamentals 1. Doug Dudgeon, Harris Group Inc. Current State of the Bank Market 2. Typical Project Finance Structure 3. Abstracts Project Structure Mitigates Project Risk 4. Project Capitalization 5. Alejandro Zamorano Cadavid Successful Financing Requires The Renewable Fuel Standard Systematic Approach 6. At Financial celebrated its seven-year anniversary Close Project Finance Execution 1. on 1 May 2013. Bloomberg New Sources and Uses 2. Private Energy Finance takes a look back at Placement Memorandum 3. privately and government-funded Independent Engineer Report 4. deals in the US next-generation Timetable 5. Project Finance Waterfall biofuels industry and appraises 6. Case Study – Project Finance Credit conversion technologies based on the Quality Stern Brothers & Co. – amount of funds invested. Alternative Energy Finance Group 1. Overview 2. Current Projects 3. Alan Propp Biographies Making good decisions is critical for companies developing and deploying Doug Dudgeon new industrial biotechnologies. In a time of limited capital availability, Decision and Risk Analysis (D&RA) is bolt-ons are all the rage in biorefining. an objective, analytical process The cash to be saved from shared designed to help companies navigate infrastructure with the host facility and through the maze of uncertainty to from smaller size can be significant make sound business and technical when compared with the cost of a decisions. D&RA allows companies to standalone installation. That said, with better understand both business and all their inherent interdependencies, technological risks so they can be bolt-on projects are necessarily more proactively managed throughout complex than standalone projects. development. With proper D&RA When considering a bolt-on companies can increase their odds of application, it is important to pursuing the right project with the remember first and foremost to do no right technology. D&RA can also be harm to the host. Additional questions used to develop a compelling, credible must be addressed in developing a Red Text: Invited 28

bolt-on application, in order to avoid unintended consequences from Advances in DOE supported Cellulosic integrating new technology with an Ethanol Projects existing, revenue-generating asset. James J. Spaeth, U.S. Department of These questions concern the Energy compatibility of core technologies, economic trade-offs between the PROESA™ technology: Break-through existing process and the new process, Technology for Producing Advanced infrastructure availability, and the Bio-Fuels and Renewable commercial affinity of the host’s Chemicals from Cellulosic Biomass current business with the potential new business. Drawing on experience Kevin Gray, BetaRenewables from Harris Group’s decades of Abstracts experience in advanced biofuels and retrofit plant design, this paper Jasmine Isar examines the critical questions of bolt- Butanol production by Clostridium ons for implementing advanced acetobutylicum ATCC 4259 was biofuels technologies in three studied using the renewable feedstock contexts: a dry-grind ethanol plant, a jatropha seed cake,. Chemical sugarcane mill and ethanol plant, and mutagenesis for improvisation of the a pulp mill. strain for better butanol tolerance and

production was done. Optimization of Tuesday, December 10, 2013 | the physiochemical parameters 2:00pm – 3:30pm resulted in about 14.3 g/l of butanol in 120 h using acid treated jatropha seed Progress in Production and cake hydrolysate (7% w/v) along with Commercialization of Alcohol AnS components. The process was Fuels scaled up to 15 L level yielding 18.6 g/L of butanol in 72 h. This is the first Moderator: James J. Spaeth, U.S. report wherein high yield of butanol Department of Energy has been obtained in single batch fermentation using acid pretreated Production of Biobutanol from jatropha seed cake. The strain was Jatropha Seed Cake found to be tolerant to 3.0 % butanol Jasmine Isar, Reliance Industries under optimized conditions and the Limited tolerance could also be demonstrated by the strain’s ability to accumulate Clean technology innovation to rhodamine 6G. The strain was found produce advanced biofuels, reduce to be having high expression of the landfilling and simulate regional stress-response protein GroEL, the economies reason behind the tolerance and Sam Park, Enerkem growth of the strain in presence of 3.0 % (v/v) butanol in the medium. Red Text: Invited 29

categories of technologies that were Sam Park developed over the last decade and The commercialization of advanced give an example of a technology using biofuels is driven by the need to non-recyclable municipal solid waste reduce dependency on oil, increase as a feedstock. The benefits of energy diversity and reduce carbon commercial-scale advanced biofuels footprint. Public policies around the facilities, as well as the advantages of world contribute to the expansion of using municipal solid waste will also biofuel production by stimulating both be presented. Finally, the presentation the supply and the demand for will explain why public policies around renewable fuels. Despite the global the world play an important role in recession, the advanced biofuels stimulating innovation and creating a industry now operates facilities, and a business climate that attracts private first wave of technologies is now ready investment to allow for the to be commercialized, as large scale commercialization of these facilities currently being built. A wide innovations. variety of biomass and residues are used as feedstock for these projects, James J. Spaeth including municipal solid waste. Three DOE will provide an update on main categories of technologies are progress and developments in DOE being developed and commercialized supported cellulosic ethanol projects. in this sector. The ones capable of using municipal solid waste (MSW) as Kevin Gray a feedstock provide an alternative to Since 2006 Biochemtex has invested landfilling and incineration, thereby approximately $200 million in the offering additional environmental development of the PROESA™ benefits. MSW feedstock has a large technology. The process is designed to potential, given its abundance, and provide low-cost, high quality second the fact that it is already collected. In generation sugars readily convertible addition to providing a new source of into bio-fuels and/or bio-chemicals. energy, advanced biofuels facilities PROESA™ integrates an energy stimulate regional economies, create efficient, chemical-free biomass high quality jobs, provide synergies pretreatment operation and a novel with traditional sectors such as viscosity reduction, enzymatic forestry, agriculture and waste, and hydrolysis step. The unique contribute to revitalize the industrial configuration ensures limited manufacturing sector. formation of degradation products that could lower yield and inhibit The presentation will talk about the (bio)catalyst performance. One of the context for the growth of this sector features of PROESA™ is the and for the use of MSW as a feedstock opportunity to process a number of for the production of advanced different biomass types ranging from biofuels. It will describe the main energy crops, agricultural residues, Red Text: Invited 30

woody biomass, and industrial by- Tuesday, December 10, 2013 | products, without the necessity to 4:00pm – 5:30pm change hardware. Biochemtex has engineered and constructed a 1 dry Overcoming Challenges in ton/day biomass processing pilot Regulation and Intellectual facility in Rivalta, Italy integrating all Property unit operations required to convert lignocellulosic biomass into fuels Regulatory Strategies for Use of and/or chemicals. BetaRenewables, a Genetically Modified Organisms in joint venture between Biochemtex, Biofuel Production TPG and Novozymes, was formed in Moderator: David J. Glass, D. Glass 2011 to license the PROESA™ Associates, Inc. technology for the production of fuels and chemicals. BetaRenewables has William Kenealy, Mascoma Corporation partnerships with leading bioconversion companies, What is patentable in the biotech Genomatica, Codexis, and Gevo, to sector? integrate PROESA™ with downstream Konrad Sechley, Gowling Lafleur processes to produce value-added Henderson, LLP chemicals. The PROESA™ technology is the basis for one of the world’s first The Here and Now of TSCA commercial scale cellulosic ethanol Regulations on Biobased Chemicals plants located in Crescentino, Kathleen Roberts, Bergeson & Italy. This plant, which opened in Jan Campbell Consortia Management, LLC 2013, is designed to produce approximately 20 MM gallons of Abstracts ethanol from a combination of agricultural residues (wheat straw) David J. Glass and energy crops (Arundo William Kenealy donax). Construction of a second Genetically modified microorganisms plant in Brazil is expected to be are broadly coming into use for the completed in Q1 2014 while similar production of biofuels or bio-based scale plants in North Carolina and chemicals. These include modified California are in the design yeast strains with improved or more phase. Lastly, M&G Chemicals efficient ability to ferment ethanol, recently announced plans to build a 1 photosynthetic microorganisms to MM ton/yr PROESA™ biorefinery produce fuels from solar energy, and producing both bioethanol and microbes that can synthesize alkane biobased chemicals in China. This fuels or fuel precursors. These are in plant will be almost four times the size addition to the numerous industrial of the facility in Crescentino. enzymes used in ethanol

fermentations that are now produced in modified production strains. Many Red Text: Invited 31

of these GMMs will need to navigate advanced yeast products which reduce the unique regulatory regimes that the cost and increase the yield of have been established in the U.S. and grain-based ethanol production. elsewhere in the world to provide TransFerm expresses glucoamylase oversight over industrial uses of (GA) during the fermentation, which biotechnology, and a growing number can reduce the cost of separately of companies have successfully purchased enzymes as well as completed the process for their novel increase the rate of ethanol strains. production. TransFerm Yield + is an David Glass will begin this panel with advanced yeast that in addition to GA an overview of regulatory also makes less glycerol thereby requirements for industrially-useful resulting in a higher yield of ethanol genetically modified organisms in the per feedstock compared to U.S. and internationally, including conventional yeast. These are the first regulations applicable to the uses of bioengineered yeast both fully modified microorganisms, algae or reviewed by U.S. regulatory plants. This will be followed by authorities and broadly-utilized in the presentations of case studies grain ethanol industry. The genetic illustrating the paths to regulatory modifications used to create approvals for under several different TransFerm and TransFerm Yield + are regulatory regimes, including built in a robust Saccharomyces approvals for use of GMMs in cerevisiae host, utilize genes from contained manufacturing and for use safe and well-characterized donor of inactivated biomass as an animal organisms, and are very stable. feed ingredient. TransFerm and TransFerm Yield + have been favorably reviewed by the David Glass will present a case study FDA’s Center for Veterinary Medicine of Joule Unlimited's successful (CVM) for inclusion in the Association navigation of the U.S. biotechnology of American Feed Control Officials regulatory system for a novel (AAFCO) official publication of feed microorganism for the production of ingredients. In addition, both products ethanol. This is a strain of have been reviewed by the EPA under cyanobacteria modified to produce the MCAN program. ethanol from sunlight, carbon dioxide and water, which was the subject of a Konrad Sechley Microbial Commercial Activity Notice The challenge to obtain patent submitted to EPA, and approved protection for technology in this sector through a voluntary consent order has never been greater. Are allowing use of the organism at Joule's nucleotide sequences still patentable? Demonstration Plant in New Mexico. Is the yeast strain we developed patentable? What data is required to Next, William Kenealy will discuss protect biotech related inventions? Mascoma’s experience with its Much uncertainty exists in the Red Text: Invited 32

requirements to protect innovation development within the biofuel and Wednesday, December 11, bioenergy sector. These and other 2013 | 8:30am – 10:00am issues that impact what is protectable, the scope of protection, and Advances in Aviation Biofuels information required to support protection of innovation will be Moderator: Michael Vevera, Mercurius reviewed within the US, Europe and Biorefining Canada. The take home messages arising from recent decisions in the Is there a renewable aviation fuel US, Europe and analogous cases in feedstock? Canada, pertaining to the types of Eric Mathur, SGB subject matter that can be protected, the extent of experimental support, Liquid Phase, Catalytic Conversion of and what is required for utility will be Biomass to Bio-Jet discussed. This presentation will Karl Seck, Mercurius Biorefining compare trends within the European, US and Canadian Patent Offices, with Alaska Airlines Investments in Aviation an emphasis the biotech sector and Biofuels how these trends impact protection of Jay Long, Alaska Airlines biofuel and bioenergy innovation. Highlights of Two Aviation Biofuel Kathleen Roberts Research Efforts by USDA NIFA and How do the evolving biobased and renewable chemical technologies fit the FAA within the current Toxic Substances Ralph Cavalieri, Washington State Control Act (TSCA) framework? The University short answer is “not well.” The longer answer involves detailed evaluation, probable notification, reporting, and Abstract recordkeeping requirements. This session will focus on helping Eric Mathur manufacturers understand the journey Drop-in replacements for petroleum their biobased product will face in the liquid fuels represent a major U.S. Environmental Protection component of the growing energy Agency’s current system of TSCA demand. Although cellulosic ethanol regulation, including potential delays, will mitigate the depletion of oil detours, and pitfalls. We will consider reserves used for gasoline and algal strategies, tips and helpful hints that biofuels hold long term promise as jet companies can use as they navigate fuel substitutes, there are no obvious the TSCA gauntlet. Without this present day renewable feedstocks knowledge, commercial success will be available. Candidates include canola, virtually unachievable. oil palm, rapeseed and soybean; yet Red Text: Invited 33

redirecting agricultural resources maximized time for accumulation of toward energy feedstocks negatively selected and unselected mutations. impacts food security and is not Jatropha planting materials were sustainable. A promising alternative is collected throughout Central America the non-edible oilseed shrub, Jatropha in order to create a germplasm curcas. Jatropha is native to Central repository which now comprises over America and was distributed by 12,000 genotypes derived from ~600 Portuguese sailors to colonies three accession families. The plant collection centuries ago in the Cape Verde exhibits high levels of phenotypic Islands. The primitive crop was diversity including variation in recognized as heating oil and as a flowering time, oilseed content, fruit result, 35,000 tons of Jatropha seed yield, plant architecture, susceptibility was exported from Cape Verde to to fungal pathogens, pest resistance, regions throughout Africa, Asia and drought, heat, flood and cold Latin America. The spread of a few tolerance.Our results now confirm that Jatropha cultivars from the center of genotypic diversity underlies the origin to the pan tropics created a observed phenotypic variation.Small genetic bottleneck in the diversity of sequence repeats (SSR) and genome- Jatropha found outside Central wide SNP markers were used to America. Recent attempts at analyze diversity of the germplasm domestication of Jatropha failed collection. The results conclusively because the plantations used demonstrate that the genomic undomesticated landraces derived variation residing in the Central from Cape Verde germplasm which American germplasm collection limited genetic improvement through positively correlates with the breeding and selection. The inability to phenotypic diversity. Moreover, the commercialize Jatropha as an energy analysis reveals that virtually all feedstock was not a reflection on Jatropha land races cluster tightly, Jatropha, but rather of the business confirming genome scale homologies getting in front of the science of and derivation from a common domestication. Here, we present ancestor. In contrast, the germplasm evidence that Jatropha curcas collected near the center of origin of germplasm is not genetically the species forms eight divergent depauperate;but rather encompasses clades, punctuated with a wide a substantial pool of genetic variation spectrum of genotypic variance within sufficient to propel breeding efforts each clade. Thus, our findings suggest designed to achieve economic yields. that Jatropha curcas possesses the It is well known that the highest genetic potential necessary for crop degree of genetic diversity within a improvement. Considering the short species is typically found near the generation time of this perennial and center of origin. High genetic diversity the ability to propagate both by sexual at the origin of a species is a and vegetative methods, there are no consequence of population density and apparent genetic obstacles preventing Red Text: Invited 34

Jatropha from becoming the preferred also provide much needed supply oilseed feedstock for renewable jet integrity. fuel. Ralph Cavalieri Karl Seck In recent years, the USDA National Mercurius Biorefiningwas established Institute of Food and Agriculture to produce profitable drop-in fuel (Jet (NIFA) and the Federal Aviation Fuel and Diesel) from non-food Administration (FAA) have biomass feedstocks. Mercurius is competitively awarded funding that developing a “faster ~cheaper~ directly supports the development of better” method of producing profitable alternative jet fuels, with emphasis on drop-in biofuels. Mercurius uses a biologically derived jet fuel. Two such proprietary, patent pending projects are being led by Washington technology called Renewable Acid- State University. hydrolysis Condensation Hydrotreating The USDA NIFA Agriculture and Food (“REACH”) to make profitable drop-in Research Initiative Coordinated hydrocarbon liquid fuels. Agricultural Project entitled Northwest

Advanced Renewables Alliance (NARA) Jay Long seeks to develop a sustainable supply This presentation describes strategies, chain that uses forest residues as the motivations, and future plans for primary feedstock and biochemically integrating aviation biofuel into normal produces jet fuel and other valuable airline operations. As aviation biofuel products. The five-year project is in its technologies evolve, driving production third year and involves about 180 rates up and unit costs down, the use individuals from multiple universities of alternative aviation fuels as part of and companies. Important progress the standard fuel sourcing operation is has been made in addressing technical drawing closer to a reality. Alaska as well as sociological and economic Airlines flew 75 flights using an 80/20 challenges. Four pretreatment blend of HEFA based bio-jet in 2011 as pathways have been examined as well part of a “proof of concept” initiative as methods for economically collecting aimed to support the aviation biofuel and transporting feedstock and production efforts. The airline is developing valuable co-products. currently engaged in a project with Industrial partners, such as Hawaii BioEnergy for a project to Weyerhaeuser and Catchlight, are produce aviation bio-fuel in an area working closely with the project’s that currently sees very high jet fuel university researchers on feedstock, prices and significant supply pretreatment, and co-product issues. constraints. It is Alaska’s opinion that Conversion of the hydrolysate to identifying these types of markets will isobutanol and ultimately jet fuel is not only allow the producers to advancing through the efforts of Gevo, compete on an economic basis, but will another of the project’s industrial partners. A parallel conversion Red Text: Invited 35

pathway to jet fuel, known as Moderator: Jim Flatt, Synthetic BioChemCat, is also being explored. Genomics Educational programming, a major objective of the NIFA program, is Marine Microalgae for a Sustainable addressing needs for STEM education Future-Production and Application of related to biofuels at all levels from Algae for Fuel, Feed and primary grades to secondary and Nutraceuticals undergraduate education. Valerie Harmon, Cellana LLC

The second project, very recently Algae, beyond energy awarded by the FAA, is the Center of Jose Sanchez, OriginOil Excellence for Alternative Jet Fuels and Environment. This new Center of Commercialization of New Algae-based Excellence (COE) is co-led by the Omega-3 Supplements with High Massachusetts Institute of Technology Bioavailability and involves a total of sixteen Brian L. Goodall, Valicor Renewables research universities. The COE also LLC engages nearly fifty companies and other organizations as research Abstracts collaborators and stakeholders. The FAA identified ten overarching Jim Flatt research objectives in the solicitation Projected growth in world population that address all aspects of and changes in dietary trends, coupled development, testing, and certification with constraints on arable land, water of alternative jet fuels as well as and carbon emissions will increasingly environmental impacts of aviation tax conventional agricultural food related to noise, emissions, and production methods and drive energy consumption. development of alternative crops which can augment traditional sources Synthetic Biology, of food and feed products. Large- Algae and Marine scale cultivation of microalgae has the potential to meet a significant portion Biotechnology of future needs, however, requires significant development to establish Monday, December 9, 2013 | cost-effective and robust production of 8:30am – 10:00am strains with desirable nutritional properties. Microalgae are relatively Optimizing Algae Production for unique in their ability to produce a Beneficial Bioproducts large fraction of their biomass as high- quality protein as well as other Microalgal-based Food and Feed nutrients including oils and carotenoid Ingredients – Crop of the Future? pigments. Researchers at Synthetic Genomics have assembled a vast and Red Text: Invited 36

diverse collection of microorganisms of commodity food and feed products. including algae collected globally SGI has recently developed algal through its bioprospecting programs. strains allowing significantly improved Hundreds of these have been light availability in a mass culture evaluated for nutritional quality, setting, without compromising leading to the identification of photosynthetic performance of the microalgal lead strain candidates strain, providing the foundation for which provide similar amino acid improved production economics. profiles for various plant and animal proteins, including soy, canola and Valerie Harmon pea, dairy proteins and fishmeal Cellana is a leading developer of proteins. Algal protein ingredients algae-based bioproducts, and its pre- must meet a number of other commercial production of marine requirements in order to be microalgae takes place at the Kona considered mainstream sources for Demonstration Facility (KDF) in human and animal nutrition, including Hawaii. KDF is housing more than 80 desirable olfactory profile, neutral high-performing algal strains for color, low allergenicity and different bioproducts, of which over 30 compatibility with different food have been grown outside at scale. The matrices, which may require either patented ALDUOTM algal cultivation aqueous soluble or insoluble proteins. technology allows Cellana to The use of microalgal biomass as a successfully tackle the issue of partial replacement for fishmeal in contamination, one of the biggest aquafeeds represents another technical challenges for algal mass promising market application. Specific cultivation in open ponds. As a result, microalgal strains provide proteins and algal mass cultivation is no longer lipids similar in composition to those limited to a few extremophiles, of current sources such as menhaden instead, we are able to tap into the as well as desired micronutrients. hugely diversified algal strains in Microalgal cultivation currently is still nature and significantly expand the too expensive and limited in scale to catalog of algal strains for mass compete on price and availability cultivation. To date, Cellana has today. However, advances in screened a large collection of algal synthetic biology and genomics offer strains and produced more than 10 powerful new tools to improve strain metric tons of biomass from numerous performance and robustness, which natural marine algal strains for the are required for attractive production development of biofuels, feed, and economics. Strains exhibiting high-value nutraceuticals. Cellana’s improved photosynthetic efficiency research and production at KDF have relative to wildtype ancestor strains addressed major areas that are crucial are required to achieve higher for the commercialization of algal biomass productivity and titer biofuels: yield improvement, cost associated with economical production reduction, and the overall economics. Red Text: Invited 37

Commercially acceptable solutions harvests algae with just finely tuned have been developed and tested for electrical pulses. In the first stage, the major factors limiting areal pulses cause the algae to coagulate. productivity of algal biomass and lipids In the second, a cloud of microbubbles based on years of R&D work gently lifts the algae to the water conducted at KDF. This talk will share surface. In 2012, OriginOil teamed up some of the highlighted R&D work, with their first customer, MBD Energy including areal yield improvement, oil to pilot the algae harvesting optimization, Omega 3 fatty acid technology, freshly patented in development, flue gas utilization and Australia. Since then the company has feed application of defatted algal expanded rapidly, recently installing biomass for both aquatic species and its technology in the La Defense livestock. Improved overall economics building complex in Paris as a part of a was achieved through a holistic dual water clean-up and energy approach by integration of high-value generation system. The team also co-products in the business model. recently completed tests validating an increase in the shelf life of the algae Jose Sanchez feedstock when harvested through Algae is one of nature’s most efficient OriginOil’s EWS technology. and versatile photosynthetic factories. Additionally, OriginOil has found that With a short growing cycle, algae is EWS can accelerate the production of not only an attractive and healthy astaxanthin by sanitizing and source for natural feed and fertilizer, concentrating the Hematococcus but also is a viable petroleum pluvialis (HP) algae for more effective alternative in fuels and chemicals. stressing. In this session, VP Jose Unlike petroleum, algae absorbs Sanchez will discuss the need in the rather than emits CO2. The main algae industry to reduce production challenge for implementation of this costs and solutions available to do so. resource is overcoming cost barriers In the second half of his presentation, associated with algae production, Mr. Sanchez will address the since often times the same amount, if burgeoning market applications for not more energy is expended high quality algae. Using the recent generating algae than is generated by OriginOil findings regarding shelf life it. Current methods for separating and astaxanthin production, Riggs will algae from water such as centrifuge educate attendees about new growth and membranes are simply too sectors in the algae industry. expensive for large-scale implementation. Centrifuge in Brian L. Goodall, co-authors Isaac particular can also damage the algae Berzin, Sam Couture, Kiran Kadam & product rendering it useless in many Brian Waibel Qualitas applications. Enter OriginOil, Health and Valicor Renewables have developer of a chemical-free, high- partnered to commercialize the first speed technology that efficiently algae-based pure EPA supplement. Red Text: Invited 38

The integrated production facility that the polar lipid composition that (from algae cultivation to finished makes Krill oil so bioavailable comes product) is located in the Southwest from the algae in their diet. Together United States and enjoys many critical Valicor Renewables and Qualitas benefits – excellent climate and high Health have developed a new patents- insolation, essentially unlimited access pending nutraceuticals platform which to brackish water, land availability, permits the efficient recovery and infrastructure and abundant local fractionation of these EPA-containing sourcing of carbon dioxide thanks to polar lipids from their naturally- the local oil and gas industries. The occurring algae source without strain used is Nannochloropsis, a hydrolysis or other degradation. natural strain not genetically engineered. The strain exhibits Monday, December 9, 2013 | exceptional EPA production capacity, 10:30am – 12:00pm has been grown in multiple locations including Israel and 2 different states New Approaches to make in the Desert Southwest. Crop Biology Easier to Engineer protection methodologies for this strain have been developed over the Reliable parts to make biology easy to last several years giving us a proven, engineer robust growth platform. Two Moderator: Vivek Mutalik, Lawrence additional major benefits of the Berkeley National Lab selected strain are the fact that it delivers EPA in the absence of other Developing designable genetic control omega-3’s such as DHA, and that the systems for applied synthetic biology bulk of the EPA resides in the biomass James Carothers, University of as polar lipids e.g. phospholipids and Washington, Seattle glycolipids. Since EPA and DHA compete for the same receptors in the High-throughput, one-pot gene body, for adults seeking the health synthesis and characterization benefits of EPA (such as anti- Tim Hsiau, University of California, depression) it is pure EPA, rather than Berkeley an EPA / DHA mix, that is important for effectiveness. In addition, the EPA Ginkgo Bioworks: A factory for dose taken is less critical than how engineering organisms much of the EPA ends up in the blood Jason Kelly, Ginkgo BioWorks stream and target organs – the bioavailability of the EPA source. In Abstracts the market today, there are products that are more bioavailable, like Krill Vivek Mutalik oil, which is premium priced and Our ability to routinely engineer enjoys market growth of 40% per genetic networks for applications is annum. It is important to remember limited by the scarcity of a well Red Text: Invited 39

characterized compendium of diverse engineer RNA-regulated genetic regulators that are orthogonal (that is, devices with quantitatively-predictable do not inadvertently cross-react), activities that can control gene homogeneous (operate with similar expression in metabolic pathways kinetics, thermodynamics and other producing industrial chemicals in E. structural properties) and have coli (Carothers et al. & Keasling. predictable functionality in variety of Science 2011). We have also contexts. There is also a need for demonstrated that models and developing methods to study the simulation tools can be used to inform performance variability of the design of microbial dynamic characterized part in a particular sensor-regulator systems (DSRS) sequence, environmental and host engineered to produce fatty acid- context, so that genome scale based chemicals and fuels (Zhang, engineering efforts can be realized. Carothers, & Keasling. Nature Biotech. 2012). At the University of In this talk, I will present a series of Washington, we are developing examples in developing large designable RNA-based systems to compendium of genetic parts that solve control problems in a 15 gene function reliably in different contexts, pathway engineered to produce methods to characterize the context substituted styrenes for advanced effects on part performance, polymer applications in photonics, sequence-activity models to photolithography and biomedicine. understand the mechanistic details of Here, I will present these results and a part operation and the development discuss the potential for using the of software tools to aid in sharing and conceptual and experimental visualization of these open source frameworks that we have established parts collections. These tools are to create full-fledged design platforms valuable in building bio-manufacturing for applied synthetic biology. platforms to produce key components of fuels, medicines and biochemicals. Tim Hsiau Cheap and reliable gene synthesis James Carothers methods will benefit the field of Naturally-occurring metabolic systems biotechnology. To address this have evolved control circuitry that problem, we have developed an minimizes the accumulation of toxic inexpensive gene synthesis and error intermediates and maintains correction method. In our system, we homeostasis. At present, engineering can make hundreds of genes in a one- even rudimentary control circuitry for pot fashion from microarray- synthetic biological systems remains synthesized oligos. We then very difficult. We have successfully developed a genetic system for formulated design-driven approaches selection of correct sequences from that use mechanistic modeling and the background pool of error- kinetic RNA folding simulations to containing genes. Our genetic system Red Text: Invited 40

can also provide information about the solubility of the synthesized gene. Farzad Haerizadeh, Life Technologies Such solubility information will be useful in ranking algorithms for Abstract rational design of genetic and metabolic pathways. Aquatic photosynthetic organisms, including cyanobacteria, micro algae, Jason Kelly diatoms and macro algae (seaweeds) Organism engineering firms are have attracted significant interest as emerging that build microbes to potential sources of specification for customers. Making photosynthetically-derived and heavy usage of robotics and carbon-neutral renewable feedstocks, computer-aided design (CAD), these specialty chemicals and biofuels. The companies are closer to application of metabolic engineering semiconductor chip fabrication plants tools and approaches to cyanobacteria than to biotech labs. Ginkgo and algae creates opportunities for Bioworks opened a new 11,000 sq ft coupling bioproduction to direct solar organism fabrication facility in energy conversion without the need early 2012. This talk will describe the for organic carbon feedstocks. Recent facility and our work advances in the understanding of designing and building microbes for biological circuitry and its regulation in the production of chemicals and these organisms has now enabled fuels -- including our development of effective application of technologies organisms engineered to use C1 for strain selection, targeted genetic compounds as feedstocks. manipulation and synthetic biology to chemicals, food, feed and medicines. Monday, December 9, 2013 | In parallel, product-optimized 2:30pm – 4:00pm cultivation and product isolation strategies have evolved rapidly as Metabolic Engineering of projects have moved toward Aquatic Photosynthetic commercialization. This session will highlight significant advances in Organisms metabolic engineering of cyanobacteria / algae as well as the Moderator: Peter Heifetz, Heifetz development of integrated and BioConsulting scalable platforms for specialized bioproduction. Mark Hildebrand, University of California, San Diego Tuesday, December 10, 2013 |

2:00pm – 3:30pm Margaret McCormick, Matrix Genetics, Inc. Updates on Algal Biofuel David Stern, Cornell University Production in China Red Text: Invited 41

known for huge diversity of aquatic Production of Biofuels and Chemicals life surprisingly only little work has Using Microalgae Isolated from the been done to use local microalgae Coastal Regions of China resources for biofuel and chemical Moderator: David Guangyi Wang, production. Recently, we have Tianjin University isolated 18 marine microalgal strains, Development of Jet Biofuels for a belonging to the families Sustainable China Aviation Industry Chlorellaceae, Scotiellocystoidaceae, Pengcheng Fu, Beijing University of Neochloridaceae, Selenastraceae and Chemical Technology Scenedesmaceae. Of isolated strains

Mychonastes afer PKUAC 9 and Synthetic photosynthetic Scenedesmus abundans PKUAC 12 Cyanobacteria to produce fine chemicals directly from CO2 were selected for biomass and ethanol Weiwen Zhang, Tianjin University production analysis. Comparison of three cultivation modes (stationary, Large-scale Production of Microalgal shaken and aerated) resulted in the Biomass and Chemicals in China: highest biomass productivity obtained Current Status and Perspective for aerated cultures that yielded 0.09 Zhongxin Yang, Hangzhou Xinwei Low-carbon Tech R&D Ltd, Hangzhou, g and 0.11 g dry weight per day per Zhejiang, P. R. China liter of medium for M. afer PKUAC 9 and S. abundans PKUAC 12, respectively. Carbohydrate Abstract accumulation monitored by FTIR

David Guangyi Wang showed that early stationary phase is Microalgal biomass has been optimal for biomass harvest. considered as one of the most Microalgal biomass was successfully promising feedstocks for biofuel (e.g., used as a carbohydrate feedstock for ethanol and biodiesel) and industrial fermentative bioethanol production. chemical production. However, Scenedesmus abundans PKUAC 12 selection of microalgal strains with was superior feedstock for bioethanol desirable physiological and production when pre-treated with the biochemical features from local combination of dilute acid treatment habitats has been one of major and cellulase. Additionally, two challenges in the large-scale strains, Chlorella Sorokiniana 21 and production of microalgal biofuels and Monoraphidium 3s35, were found to chemicals. Although the coastal areas produce large amount of long-straight of Pearl River Delta have long been chain alkane and fatty alcohols under the optimized cultivation conditions Red Text: Invited 42

and can be used to produce important practices and potential competition supplements for petrodiesel-like fuels with global food resources, while and chemicals used in the cosmetics helping to lower aviation carbon dioxide outputs. Details will be and food industries. discussed. Pengcheng Fu Viable and alternative fuels for Weiwen Zhang commercial aviation uses can play an Photosynthetic Cyanobacteria have important role for the emission attracted significant attention as a reduction and sustainable economic “Microbial factory” to produce biofuels development. PetroChina and Boeing and various fine chemicals, mostly due are leading respective teams in China to their abilities to utilize CO2 and for exploring second-generation sunlight directly as carbon and energy biofuel feedstocks and processes that sources, respectively. We will report have the potential to reduce our recent progress on engineering greenhouse gases throughout their cyanobacterium Synechocystis sp. entire lifecycle. PCC 6803 to produce 3- hydroxybutyrate (3HB) and butanol Three interactive efforts have been directly from CO2, and on made in order to develop a strengthening Synechocystis for better sustainable aviation fuel industry in tolerances against toxic products. China: 1) Industrial evaluation of feedstocks, processing and Zhongxin Yang demand/distribution for aviation The focus of my presentation is the biofuels; 2) demonstration flights and industrialization possibility of 3) Supply-chain R&D focused on algae microalgae as a fuel in China. From a feedstock and processing techniques. business perspective, the cost is the main limiting factor to the microalgae The study may result in the fuel industrialization. The cost consists environmentally progressive fuel of the following four technical aspects: solutions (bio-jet fuels) that differ algal species, cultivation site and from traditional biofuel development. cultivation technique, harvesting and Jet bifuels will incorporate second- extracting technology, converting generation methodologies relative to technology and final products sustainable feedstock source selection application. Furthermore, my and fuel processing, which are presentation will explore the uniquely suited for aerospace possibility of algae cultivation in applications. These jet biofuels can various regions in China, as grouped potentially be blended with traditional by energy adequacy, water and land kerosene fuel (Jet-A) to reduce availability, the abundance of carbon dependency on petroleum-based fuels. dioxide, the suitability towards factory Additionally, sustainable feedstock farming, and so on. To be considered sources may avoid deforestation the most ideal location for microalgae Red Text: Invited 43

biofuel production, the following Abstracts criteria have to be met: 1)There has to be an abundance of energy, i.e. a Sunil Chandran large variance in temperature Amyris is building an integrated throughout the day, and algae renewable products company to apply cultivation can be maintained industrial synthetic biology to throughout the year in nature genetically modify microorganisms, condition; 2) Virtually no rainy days, and more clear days. This is conducive primarily yeast, to serve as living to year round algae farming; 3) factories. These modified yeast strains Adequate water resources, including convert plant-sourced sugars into fresh water, salt water, waste water, potentially thousands of isoprenoid and sea water; 4) The availability of molecules, providing a broad range of economical land as granted for such renewable chemicals and purposes. transportation fuels. Synthetic biology Tuesday, December 10, 2013 | approaches at Amyris have 4:00pm – 5:30pm accelerated creation and improvement of S. cerevisiae strains that make high Biosynthetic Pathways to levels of isoprenoids. This presentation Desired Renewable Chemicals will describe our integrated yeast strain engineering and phenotyping An industrial biotechnology pipeline for pipeline and its application for optimizing isoprenoid production in combinatorial exploration of thousands Saccharomyces cerevisiae Moderator: Sunil Chandran, Amyris of complex genotypes to continuously improve isoprenoid-producing strains Systematic level engineering E. coli over the past five years. and S. cerevisiae efficiently producing aromatic amino acid and ethanol Liu Shuangping, co-authors Liang Liu Shuangping, Jiangnan University Zhang, Guiyang Shi National Engineering Laboratory for Renewable Chemicals from Waste Cereal Fermentation Technology Carbon (NELCFT) is an advanced biomass Lisa Dyson, Kiverdi biotransformation research center in

China. Some microorganisms Clarification of succinic acid fermentation broth by ultrafiltration producing organic acids and ethanol and the associated membrane fouling are being building by applying mechanism industrial synthetic biology in NELCFT. Jianmin Xing, Institute of Process In microorganisms, the control Engineering, CAS architectures of the biosynthesis pathway for desired chemicals always Red Text: Invited 44

form a system, it is difficult to release glycerol formation, cellobiose and all the drawbacks generated by high protein utilization, and intracellular intracellular concentrations of desired redox balance. chemicals or intermediates. With the development of contributing tools Lisa Dyson (hosts, vectors, genetic controllers, Kiverdi is an advanced sustainable oils and characterized enzymes) in and chemicals company developing bio-based replacements to synthetic biology, the costs and time oleochemicals and petrochemicals are decreased to develop an effective using our proprietary Carbon cell factory at systematic level. The Engineering Platform, a bioprocessing biosynthesis of aromatic amino acids technology capable of converting low- (like phenylalanine, tyrosine and cost, abundant waste carbon in form tryptophan) is one of the most of syngas or industrial waste gas into complicated amino acid synthesis high-value renewable chemicals at a fraction of the cost of current pathways. In this study, the approaches. Kiverdi’s technology uses engineering of a L-phenylalanine (Phe) a class of proprietary producer was carried at systematic chemoautotrophic microbes that act level: modify the PTS system to as “whole cell” biocatalysts in our moderate the glucose uptake rate innovative 1-step bioprocess for the decreasing the acetic acid formation; direct production of renewable organic genetic switch on or off the expression molecules from gaseous inorganic carbon and hydrogen for a wide of target genes to conciliate cell variety of product applications such as growth and precursors synthesis; surfactants, polymers and fuel attenuate the shunting pathway to additives. construct non-growing cells; and also increase the enflux by expression the With increasing levels of waste exporter. After these works, an generation and decreasing available space for disposal, each year 134M industrial E. coli with high productivity tons of landfill, 170M tons of and yield was constructed which was agriculture and 60M tons of forest comparable or even better than waste generated in the United States existing producing strains. Based on is an untapped biomass resource to the chassis above, the organisms produce high value chemicals. producing D-phenylalanine, L- Kiverdi’s solution fills a “scale gap,” phenylglycine and tryptophan are that enables localized, low CapEx deployment of waste-derived syngas being constructed. Besides, our team conversion plants to target diverse also constructed a super S. cerevisiae chemistries to make drop-in and at the system level with the abilities of custom renewable chemicals with mid higher permeability resistance, lower to long carbon chain lengths, varying Red Text: Invited 45

levels of unsaturation, and added the main factor that caused the functional groups that serve as operation resistance. Results showed sustainable intermediates for that most membranes selected in this surfactants, polymers or fuel additives study tended to be fouled by cake that can compete on cost and layer or concentration polarization. performance. Our feedstock flexibility Hermia’s model, which is composed of and high-yield bioprocess enables us four individual sub-models, was used to produce drop-in and custom to analyze the predominant fouling renewable chemicals allowing us offer mechanism for the used membranes. downstream customers predictably Results showed that the fouling of RC higher margins and supply certainty 10kDa and PES 30kDa was controlled for achieving their sustainability goals. by the complete blocking mechanism, while PES 100kDa was controlled by Jianmin Xing the intermediate blocking and PES Ultrafiltration was investigated to 10kDa was controlled by cake layer. clarify succinic acid fermentation broth This conclusion was also proved by in consideration of integrating SEM photos. Membrane characteristics fermentation and separation process were analyzed before and after and removal the product in situ. ultrafiltration by AFM and goniometer. Different membranes(PES 100kDa, Both contact angle and roughness of PES 30kDa, PES 10kDa and RC most membranes increased after 10kDa) were used and two models ultrafiltration. were applied to analyze the fouling mechanism. Results indicate that Wednesday, December 11, ultrafiltration is feasible in clarifying 2013 | 8:30am – 10:00am succinic acid fermentation broth. Almost all the microorganism cells Techno-economic and Physical (99.6%) were removed from the Analysis for Microalgae Growth fermentation broth. Proteins were also removed effectively by all membranes Moderator: David Hazlebeck, Global selected in this study. The removal Algae Innovations rate was 79.86% for PES 100kDa,

86.43% for PES 30kDa, 86.83% for Algal Lipid Trigger and Grow Tank PES 10kDa, and 80.06% for the RC System 10kDa. After ultrafiltration, clearer Mark Randall, T2e Energy, LLC permeate was obtained comparing with the centrifugation. Among all Techno-economic and Fluid Dynamics tested membrane, PES 10kDa gave Analysis for Growing Microalgae with the best results which showed relative the Intent of Producing Biofuel Using a higher initial flux (18.86 L/m2 h), System Model highest protein removal rate (87%), Leah Raffaeli, University of Denver and less flux drop rate. Resistance-in- series model was applied to determine Red Text: Invited 46

A Dynamic Imaging Cell Monitoring enables T2eH to produce a predictable System for Real-Time Analysis of level of oil and biomass production Algae-to-Biofuel Production with a symbiotic modular system co- Victoria Kurtz, Fluid Imaging located near existing coal, gas, or Technologies, Inc. wood fired power plants. With this approach T2e can produce a gallon of Algae Testbed Public Private algal oil at scale for less than Partnership (ATP3): An Experimental $1.00/gallon anywhere in the world. Framework for Performing Long Term Cultivation Trails Across Different The presentation will cover results Regional, Seasonal, Environmental, from UCSD’s Center for Algae and Operational Conditions Biotechnology, US Department of John McGowen, Arizona State Agriculture and Ohio State’s Ohio University, AzCATI BioProducts Innovation Center that verify the level of algal growth Abstracts including total biomass, lipids and the resulting change in oil profile. We will Mark Randall also discuss our unique methodology T2e Energy Holdings ("T2e") is a start- for our mixotrophic system that up alga to biofuels company located in includes our patented algae grow tank San Diego dedicated to use its system that enables T2e to grow algae patented and third party verified adjacent to existing power plants LipiTriggerTM technology to produce anywhere in the world. high value algae products. T2e’s Leah Raffaeli LipiTriggerTM and grow tank Techno-economic and systems studies technology not only increases algae to date are abbreviated and missing a biomass productivity and lipid content number of important variables. By but also improves algae lipid profiles including these variables in a detailed favorable for biofuels model integrating biology, chemistry, production. T2e's novel intellectual engineering, and financial aspects, a property uses a highly sustainable, more defined fluid dynamics and waste resources-based financial analysis are possible. mixotrophic approach in conjunction Through optimizing the model with our patent pending grow tank productivity based on the resulting net system to provide certain strains of profit, the system analysis results in a algae with the most optimal controlled more accurate assessment of growing environment. This approach environmental and economic has achieved unequalled results sustainability of specific algal growth including greater algal density while scenarios. Photobioreactor algal favorably changing the oil profile that growth scenario optimization in the serves the biochemical/bioplastics and system model has resulted in realistic biofuels industry. Controlling every engineering design requirements. variable of the algal growth process Results show feasibility for Red Text: Invited 47

photobioreactor growth scenarios to concentrations are produced in real- be economically sustainable when co- time, and are used for trend analysis. products are included, but definite This system can be hooked into any technological advancements and part of the production flow loop for growth improvements must be made. analysis at any point in the process. The main factors inhibiting a cost The system is Class I, Div. I effective photobioreactor growth compliant, and automatically cooled to scenario are culture density, maintain proper working temperature temperature, and lighting distribution on-site. Using a unique auto-dilution for solar lit photobioreactors, and system, the concentration is adjusted lighting cost for artificially lit for optimum presentation of the photobioreactors. Open pond algal microorganisms to the imaging growth scenarios do not show any system. Since every particle image prospect of economic or and its measurements are saved by environmental sustainability with the system, it creates an ironclad current technology due to the large audit trail for how data was recorded. amount of surface area required, Test data collected in the field will be inefficient water use, and low culture shown illustrating typical results from density. All algal growth scenarios are the system. A short video will show inferior to petro-diesel regarding how the system works in real-time, energy inputs, carbon emissions, and including how the particle images are environmental sustainability. No algal acquired and measurements made. growth scenarios included in this study Finally the results of the analysis will come close to meeting the U.S. be shown, detailing how the system requirement of biofuel emitting at can be used to monitor microorganism least 20% less carbon emissions than size and concentration, and in diesel from crude oil. particular, identification of predators.

Victoria Kurtz John McGowen This presentation will detail a novel ATP3 is made possible by a $15 new system (patent applied) for real- million U.S. Department of Energy time analysis and monitoring of algae competitive grant from its Bioenergy production. The system uses in-flow Technologies Office. Our vision is to digital imaging to capture images of establish a sustainable network of all representative cells or other regional testbeds that empowers microorganisms in photo bioreactors knowledge creation and dissemination or raceway ponds. Sophisticated within the algal research community, image processing algorithms are used accelerates innovation, and supports in real time to segment each growth of the nascent algal fuels microorganism from the background, industry. Our goal is to create a and record over 30 size, shape and network of operating testbeds, gray-scale measurements for each bringing together world-class microorganism. Cell size and scientists, engineers and business Red Text: Invited 48

executives to lead the effort to achieve our goals, ATP3 is increase stakeholder access to high implementing an experimental quality facilities by making available framework named “Unified Field an unparalleled array of outdoor Studies” (UFS). cultivation, downstream equipment, and laboratory facilities tightly Through our UFS framework, we will managed by a multi-institutional and generate data for the assessment of transdisciplinary team. the current and future state of technology informing modeling efforts ATP3 is utilizing that same powerful to establish economic and combination of facilities, technical sustainability metrics and project expertise, and management structure future targets for algal biofuel to support DOE’s TEA,LCA and production. System and scale resource modeling and analysis variation has the potential to induce activities, helping to close critical unwanted non-geographical-related knowledge gaps and inform robust variability between testbeds as a analyses of the state of technology for function of differences in system algal biofuels. Anchored by the design and scale of operation between existing 300,000L, open testbed sites. In Phase 1 of our project, this facilities at ASU and augmented by variation is being minimized at the five university and commercial facilities in testbed sites via the adoption of a Hawaii (Cellana), California (Cal Poly uniform design of outdoor raceway San Luis Obispo), Ohio (Touchstone pond systems with respect to: size, Research Laboratories), and Georgia geometry; volume; depth and (Georgia Institute of Technology), hydrodynamic mixing. Identical ATP3 provides comprehensive outdoor raceway ponds have been cultivation and harvesting facilities installed at each site using a standard along with the required experience of geometry, paddle wheel design, and operating an open testbed system. monitoring and control systems. As it is also essential that each site A primary objective is to utilize our operates to a standard set of protocols team’s expertise and world-class with respect to experimental study facilities to perform long term timing and duration, operating cultivation trials with process and conditions, as well as sampling and statistical rigor, producing robust, analytical protocols for a fixed set of meaningful datasets across different parameters, we have spent Phase 1 regional, seasonal environmental and harmonizing protocols across all sites. operational conditions. These data are Without these controls for our critically important to support TEA and cultivation systems and protocols, the LCA activities that will guide research inter-site variability would be more and development towards the difficult if not impossible to interpret. transformative goal of cost- For this presentation, we will present competitive algal biofuels by 2022. To current progress on the initial Red Text: Invited 49

experimental design framework, our others. The reason for the yield loss is harmonization and alignment activities that phospholipids are emulsifiers and and results from our initial cultivation drag oil with them as they are trials. removed. Enzymatic degumming with Purifine® PLC is a unique process that Technical and Research results in increased yield of degummed oil by reducing heavy Presentations phase oil losses and release of the DAG component of the gums. The Monday, December 9, 2013 | value of implementing Purifine PLC 8:30am – 10:00am degumming is further enhanced by integration into downstream refining Development of New Industrial (for edible oil or biodiesel end uses) Enzymes and improvements in quality of co- products such as meal. Enzymes for degumming of vegetable oils are The biotech (r)evolution in oilseed hardly a new thing, having first been crushing & refining introduced to the Moderator: Tim Hitchman, DSM industry in the early 80’s. Since then,

a steady stream of technical Computational Design: Enabling New publications has demonstrated that Products, New Enzymes, and New degumming using enzymes is feasible Pathways and can be applied to deliver oil that Eric Althoff, Arzeda Corp. exceeds standard quality

specifications. Only in recent years, Developing novel strategies on laccase the economic benefits of enzyme production and purification: Recent degumming have also been quantified, advances and new trends those processors who have taken the Feng Wang, Institute of Process lead in adopting the technology for Engineering, CAS degumming of

crude soybean oil are now An Industrial Scale Platform for documenting yield gains of up to 2%. Enzymes and Other Proteins These data now allow the industry to Danai Brooks, Dyadic International, establish benchmarks for overall Inc. economics of investment for future

implementations. This practical case Abstracts about technology developments in the

oilseed crushing & refining industry Tim Hitchman will illustrate how to fast forward Degumming is the removal of evolution into revolution and provide phospholipid impurities from oil. This direction for successful biotech step results in the majority of loss of commercialization. oil yield associated with purification of crude soybean and canola oils, among Red Text: Invited 50

Eric Althoff biosynthetic pathways for the The recent emergence of industrial industrial scale synthesis of high value and synthetic biotechnology has the chemicals. potential to radically transform the chemical industry. Despite significant Feng Wang success, the availability of efficient Laccase belongs to a group of copper- biocatalysts is one of the major containing blue oxidases with the limitations to design novel cell capability of catalyzing one-electron factories that can produce valuable oxidation coupled to the reduction of chemicals renewably. To this end, we molecular oxygen to water. Because developed novel computational laccase can oxidize phenolic methods and applied them to compounds, aromatic amines, and rationally engineer enzymes with a even nonphenolic substrates in the wide range of activities, including the presence of redox mediators, it is de novo design of enzymes catalyzing widely used for various purposes such a retro-aldol reaction, a Kemp as pulp delignification and bleaching, elimination reaction, and a Diels-Alder wastewater treatment, dye reaction. To complement our previous decolorization, food processing, techniques, we have recently biopolymer modification and developed a new algorithm, called biosensors. However, low enzyme Enzyme Identification™, to rapidly yield in laccase production and high- engineer enzymes with known cost in laccase purification limit its use catalytic mechanisms for non-native in industrial applications. Great efforts substrates. For a given chemical have been made to enhance laccase reaction, our algorithm screens “in production, involving isolation and silico” large databases of structural breeding of high-producing strains, and sequence information to allow the medium optimization, inducers rapid discovery of existing enzymes utilization, reactor design and that possess both the necessary recombinant expression of laccase catalytic machinery and an genes in different hosts. Methods for appropriate substrate binding pocket. laccase purification have been For those enzymes that possess the developed for different microbial necessary catalytic machinery but lack species, including ammonium sulphate an active site that can accommodate precipitation, ultrafiltration, three- the substrate(s) of interest, Enzyme phase partitioning, gel filtration, ion Identification automatically redesigns exchange, and affinity and remodels the active site pocket chromatography. It is crucial to thus enabling catalysis of the desired improve laccase production and reaction. The demonstrated success purification for the high demand of and wide applicability of our methods laccase from industry. In this paper, open the way for the design of a we critically review recent advances in variety of novel biocatalyst necessary laccase production and evaluate the for the efficient development of advantages, drawbacks, and Red Text: Invited 51

scalability of developing technologies Study on anaerobic fermentation of for laccase purification. In addition, different organic wastes for biogas novel strategies for the enhancement production of laccase yield and magnetic Priyanka Gupta, Birla Institute of separation methods for simple and Technology, Mesra, Ranchi efficient purification of laccase are put forward according to our recent Production of ethanol from pine achievement on laccase research. needles using different fermentation strategies Danai Brooks Parvez Singh Slathia, Shri Mata What is C1? A robust and versatile Vaishno Devi University fungal platform for gene discovery, expression and the production of Continuous Fermentation: Prospects enzymes and other proteins. Based on for Future Rollout the Myceliophthora thermophila Joshua Velson, Nexant fungus, a soil-borne saprophyte that secretes cellulases. Developed in the Abstracts early 1990’s through a fortuitous UV- induced mutation and continuously William Kenealy bioengineered since. Addresses the Mascoma has developed advanced critical bottlenecks of protein yeast products for grain-based ethanol discovery, development, scale-up and production. By expressing starch- commercialization. Enables new degrading enzymes in situ during the product introduction with less time, fermentation, TransFerm™, can cost and risk. Enables new product reduce the cost of separately introduction with less time, cost and purchased enzymes as well as risk. Broad platform capabilities increase the yield and rate of ethanol validated through 17 years of R&D production. The genetic modifications and 15 years of product sales and used to create TransFerm™, are built partnerships with key players. in a robust ethanol production strain, resulting in a yeast product that is Monday, December 9, 2013 | truly a “drop-in” solution for ethanol 10:30am – 12:00pm producers. Transferm™ Yield + is an improved strain that also makes less Stabilization in Fermentation glycerol thereby resulting in a higher yield compared to conventional yeast. Processes The increased yield was enabled by

introduction of an alternative New Yeasts for Grain Based Ethanol metabolic pathway designed to Moderator: William Kenealy, Mascoma complement the reduction in glycerol. Corporation The in situ production of glucoamylase

also reduces the concentration of glucose present initially during Red Text: Invited 52

fermentation. As a result, transferm Yield + is able to rapidly produce Parvez Singh Slathia ethanol while producing less glycerol Today most energy demands are met in response to osmotic stress. The by non-renewable energy resources combination of these interactions that may lead to resource depletion provides the basis for ethanol yield and environmental problems. A improvements of up to 5%. Mascoma demand to develop novel renewable has teamed with industry leaders ICM energy harvesting technologies is and Lallemand Biofuels and Distilled urgent and the production of ethanol Spirits to bring TransFerm products to from biomass has been focused upon the ethanol production industry. recently. Most ethanol has been produced from starch based plants Priyanka Gupta causing high food and feed prices. Priyanka Gupta, Raj Shekhar Singh, Ethanol from lignocellulosic biomass Ashish Sachan, Ambarish S. Vidyarthi such as wood, agriculture wastes, Current environmental problems have energy crops, wastes from pulp and led to an increased interest in paper industry is seen as future anaerobic digestion of solid wastes. technology. Solid wastes can be stabilized by the process of anaerobic digestion thereby In the current study we have tried to generating biogas, which can be used use pine needles from Pinus roxburghii as renewable energy source. Biogas is as a source for ethanol production. a source of energy which will leave Pines are coniferous, evergreen, less of an environmental footprint, as resinous tree belonging to the genus compared to coal or oil. The objective Pinus of the family Pinaceae. Pine of this study was to see the needles from Pinus roxburghii were performance of the selected organic obtained from forests of Tikri, wastes in a single stage anaerobic Udhampur, J&K (India). The Pine reactor for biogas production. In this needles were dried and comminuted study, experiments were conducted to paas through a sieve of 2 mm. using different organic wastes i.e. Different concentrations of acid vegetable waste, garden waste, saw (H2SO4)and alkali (NaOH, KOH, dust, fruit wastes, and sugarcane aqueous NH3)were used in bagasse, under anaerobic conditions. conjunction with steam and pressure Mine water (from Jitpur colliery, India) for varying time intervals for was used as inoculum for the biogas maximizing the sugar release. The production. The study was carried out biomass stream after pretreatment in a specially designed laboratory was filtered and pH adjustment was scale reactor for a period of two made in the filtrate. The solid fraction months. Amongst the considered was dried and again treated with biomasses the highest methane yield steam for obtaining sugar from this of 212.87cc was observed with fraction. Enzyme treatment of both vegetable waste. the fractions was done using Red Text: Invited 53

commercial cellulase and pectinase Continuous fermentation is an oft- enzymes and optimization of time for cited but rarely realized goal for maximum sugar release was done. commercial biotech Enzyme loading of 5U/g of biomass processes. Despite the possible was used. For fermentation benefits of continuous fermentation Saccharomyces cerevisiae and configurations, the critical problems in Kulyveromyces marxianus were used maintaining steady state production in two different modes SSF with living organisms introduce many (Simultaneous Saccharification and risks with scale. This presentation will Fermentation) and SHF (Separate present a brief overview of the history Hydrolysis &Fermentation). Ethanol of continuous fermentation, its concentration was measured by Gas advantages and disadvantages, and chromatography at regular intervals. the prospects for the future, focusing on emerging process configurations, In our study acid treatment proved to how they mitigate many of the be better in releasing sugar than problems with older designs, and alkaline. Both the methods were used finally which bioproducts are most to carry out the further study. Enzyme likely to see commercialization of treatment with the both the enzymes continuous fermentation processes in combined gave better sugar yield the future. Designs including free-cell when enzymes were used in isolation. chemostats, immobilized cell In fermentation studies, it was seen bioreactors and membrane-integrated that SSF proved to be a better method bioreactors will be discussed. yielding more ethanol concentration than SHF. While using biomass loading Monday, December 9, 2013 | of 5g in the pretreatment processes 2:30pm – 4:00pm fermentation yield of .2g of ethanol was obtained as the best production. Advancements in Biological Further, Kulyveromyces marxianus Processes in Industrial was a better producer of ethanol Biotechnology having a higher conversion efficiency of 52% than that of Saccharomyces Moderator: Bhima Vijayendran, cerevisiae which is 47%. The biomass Redwood Innovation Partners, LLC residue left after first pretreatment has considerable amount of sugar Performance of a Carboxydothermus which was used for ethanol hydrogenoformans gas lift reactor for concentration. Thus, pine needles are syngas upgrading into hydrogen an attractive source for ethanol Mathieu Haddad, University of production and technology needs to be Montreal developed for their use in fuel ethanol industry. Thermophilic Consolidated

Bioprocessing of Cellulosic Materials to Joshua Velson Biohydrogen Red Text: Invited 54

Lew Christopher, South Dakota School cost efficiency and environmental of Mines and Technology-CBRD attractiveness.

Synthetic Biology - A Tool for Self Of interest to this study is the obligate Replicating In Situ Resource Utilization anaerobic Carboxydothermus John Cumbers, NASA Ames Space hydrogenoformans, an extreme Portal and SynBioBeta thermophilic (70°C) bacillus that uses CO as sole source of carbon and energy and catalyses the WGS Abstracts reaction.

Mathieu Haddad, co-author Serge R. We investigated and optimized, in a

Guiota 35 L gas-lift reactor, H2 conversion of CO by a pure culture of C. Gasification of biomass and waste is a hydrogenoformans. The reactor was way to produce non-fossil hydrogen operated with a continuous supply of

(H2) through high temperature (750– gas for 3 months. Reactor 1500°C) conversion of carbonaceous performance was evaluated under material into a synthesis gas (syngas) various operational conditions, such mainly composed of carbon monoxide gas recirculation (0.3 and 1.5 L/min),

(CO), carbon dioxide (CO2) and H2. CO loading rate (from 0.05 to 0.46 -1 –1 The final calorific value of syngas, that mol.L reactor.d ) and nutrient addition is its H2 to CO ratio, varies depending to the medium. Overall, results on the feedstock type used and indicated a constant H2 yield of –1 gasification conditions. 95±1% and 82±1% (molH2·mol CO) in a supported and unsupported growth One typical way of achieving medium respectively regardless of the augmentation of the H2 content of operational condition tested. Once the syngas in industry is through the use biomass concentration was sufficiently of the water-gas shift (WGS) reaction: dense, a maximum CO conversion –1 -1 CO + H2Ov → CO2 + H2. The WGS is activity of 0.17 molCO.L reactor.day or -1 catalyzed using catalysts that are 3.79 LCO/Lreactor.day was achieved. highly intolerant to sulfur and need One major parameter that impacted regeneration, and the process is both biological activity and volumetric energy-intensive, namely for steam mass transfer was the gas generation. recirculation/CO feed ratio. We here demonstrate that, as long as this ratio Alternatively, biologically-mediated is higher than 40, mass transfer WGS reactions possess several limitations are bypassed resulting in advantages over conventional maximum conversion efficiency chemically mediated WGS (90.4±0.3%) and biological activity –1 mechanisms; such as enzyme (2.7±0.4 molCO.g volatile suspended solid –1 specificity, improved yields, overall (VSS).day ). Red Text: Invited 55

whereas MCC produced 9.4 mmol Lew Christopher H2/g cellulose at 7-fold higher H2 An environmentally friendly and production rates than SWG. The potentially viable alternative for consolidated bioprocessing (CPB) sustainable H2 production is presented capabilities of C. saccharolyticus through the utilization of renewable present opportunities for cost savings cellulosic materials. Switchgrass of capital and operational expenses in (SWG) is viewed as one of the most excess of 50%. The advantages of promising energy crops for the U.S. utilizing thermophilic microorganisms conditions with low nutrient/water on low-cost carbon feedstocks without requirements and high adaptability to prior thermo-chemical treatment for any weather/soil/land conditions. bioH2 will be discussed. Production of SWG is projected to increase ten-fold within next ten John Cumbers years. Furthermore, its high Gravity is the force of nature that carbohydrate (>65%) and low lignin drives the cost of space exploration (<20%) content favors a fermentation and our eventual settlement of the route for SWG utilization. On the other solar system. To send a kilogram of hand, the handling and disposal of mass from the Earth into low earth municipal solid waste (MSW) is of orbit currently costs between $5,000 growing global concern. In the U.S. to $20,000 depending on the launch only, MSW recently reached 5 lb per vehicle. Every human breathes out capita per day. However, MSW one kilogram of CO2 every contains approximately 60% of day. Currently we do not have biodegradable material which can be technologies capable of fully closing utilized for bioenergy production. The the CO2 loop in space, meaning that H2 production capabilities of the we are wasting valuable resources. extreme thermophile Caldicellulosiruptor saccharolyticus In situ resource utilization (ISRU) is a DSM 8903 were examined on SWG set of technologies developed by and MSW, with glucose and NASA and others for reducing the cost microcrystalline cellulose (MCC) of access to space by utilizing the serving as references. It was resources on other planetary bodies demonstrated that C. saccharolyticus and in closed loop life support can ferment SWG bioH2 in a single systems. Biology offers an advantage step at 11.2 mmol H2/g without any over physical and thermal systems physicochemical or biological because a small initial mass can be pretreatment. In comparison, the sent into space and replicate into a bioH2 yields on MSW were much larger mass from the resources approximately 4-fold lower. BioH2 around it. production from glucose reached the theoretical maximum for dark In this talk I will outline a new concept fermentation of 4 mol H2/mol glucose, of resource utilization for planetary Red Text: Invited 56

sustainability. I will then outline the Abstracts resources that are available in different space environments and Brian M. Baynes discuss some of the technologies that Solid catalysts, despite their broad could be used for the production of applicability as the workhorses of the food and other consumables that petrochemical industry, have seen could be made there. little penetration in biomass hydrolysis, because the required catalytic power, lifetime, and costs Tuesday, December 10, 2013 | have been difficult to achieve. Here 2:00pm – 3:30pm we introduce a family of novel, chemically-defined materials that are Feedstock Conversion highly potent and reusable biomass Technologies hydrolysis catalysts. Mild operating temperature, relatively short

residence time on the order of an Novel Solid Catalysts for Production of hour, and easy catalyst reuse combine Renewable Sugars and Chemicals from to enable biomass to be converted to Lignocellulosic Materials sugar and other products at very low Moderator: Brian M. Baynes, Midori cost. Product selectivity can be Renewables, Inc. tailored to yield (1) fermentable

sugars, (2) soluble fiber, or (3) sugar Lignocellulosic biomass as potential dehydration products such as substrates for the white biotechnology HMF/furfural. This talk will highlight Joachim Venus, Leibniz Institute Midori’s work in invention and Agricultural Engineering (ATB) development of this technology

through 100 kg/hr pilot tests over the Characterization of an adapted past four years and its potential to microbial population to the make ultra low cost sugars and other bioconversion of carbon monoxide into renewable products a reality. butanol using next-generation sequencing technology Joachim Venus Guillaume Bruant, National Research Renewable feedstocks (e.g. crops, Council Canada lignocellulosics, green biomass,

residues) can be utilized directly, e.g. Evaluation of second generation as energy carriers, as packaging biofuels production from native materials, as fibres, for the production halophytes by chemical- of colouring agents or as lubricants. characterization of Salicornia sinus- However, they can also be converted persica biotechnologically by enzymes and Alassali, A., Cybulska, I., Thomsen, M. microorganisms, giving us access to a H., Masdar Institute of Science and multitude of new, biocompatible Technology products and possible uses. Often the

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economy of bioprocesses is still the technologies of renewable materials. problem because in the case of bulk For that purpose a multifunctional products the price is affected mainly pilot plant was planned and built at by raw material costs. The production the site of ATB to investigate different of environmental friendly, climatically raw materials and products. The sustainable basic chemicals based on construction of a pilot facility for the renewable raw materials can help to production of lactic acid from save limited fossil resources and to renewable resources consequently fills increase the economic potential of a gap in the various phases of rural areas. The cultivation and bioprocess engineering from applied utilization of renewable resources for fundamental through application the non-food application become an research to the launch of alternative source of revenue in biotechnological processes in practice. agriculture and forestry. Even though First results of lactic acid fermentation the manifold potential is used only for in a 450-L-bioreactor will be a small part today the production and presented. Depending on the further processing of renewable resources will processing of the lactic acid the be a growing economic factor in separation of impurities after future. The sugars after the pre- fermentation is a major process cost treatment of several agricultural and too. Therefore an optimization is forestry feedstocks can be converted necessary to find a balance between by lactic acid bacteria to produce lactic the substitution of expensive nutrients acid. Lactic acid, its salts and esters and the limitation of interfering or have a wide range of potential uses undesirable components of natural and are extensively used in diverse raw materials respectively. fields, e.g. bioplastics. The goal is to Exploitation of high quality L(+)- and develop a fermentation process based D(-) lactic acid for the production of on the substitution of expensive biopolymers is one of the recent nutrients and supplements by cheaper applications. Conventional processes materials from renewable resources for down-streaming are based on due to their main proportion of the precipitation steps that generate large whole process costs. Besides the basic amounts of chemical effluents. research projects respecting the Consequently the environmental screening and characterization of impact of traditional processes can be microorganisms, phenotypic reduced by using alternative optimization, down-stream processing technologies, such as electrodialysis of fermentation products, application with monopolar and bipolar and refining of lactic acid, economic membranes. assessment of bioconversion processes the scale-up to a technical Guillaume Bruant scale of several processing steps have Microbial production of butanol is still to be developed for transferable conventionally based on the utilization solutions of bioconversion of carbohydrates as carbon feedstock. Red Text: Invited 58

More and more companies are experiments. Bacterial and archaeal interested in using alternative carbon 16S rRNA genes were amplified and sources, such as biomass, notably by then sequenced using the Ion Torrent combining conversion of biomass into sequencing technology. syngas via gasification and microbial fermentation of syngas components. Data generated were analyzed using To date, only few syngas-fermenting the ribosomal database project (RDP) microorganisms that can produce Classifier web tool. Notable differences butanol are known. Discovering new were observed in the microbial microorganisms or microbial consortia community structure between the capable of fermenting syngas into initial anaerobic microbial consortium liquid biofuels, and engineering them and the CO-adapted population to make them commercially attractive obtained after enrichment is primordial in a strategy to develop experiments. Diversity of the bacterial an economically viable platform for population notably decreased after CO biobutanol production. enrichments, with 17 different phyla identified in the initial sludge, as This presentation will discuss the use opposed to 14 after one month and 11 of next-generation sequencing after two months of operation. technology to perform microbial Bacteroidetes, Actinobacteria and community analyses of anaerobic Proteobacteria, which were the undefined mixed cultures, with the dominant phyla in the initial sludge, objective to identify microbial species significantly decreased after CO particularly adapted to the enrichments. On the opposite, bioconversion of carbon monoxide Firmicutes became ultra dominant, (CO), a major component of syngas, representing more than 82% of the into butanol. CO enrichment total bacterial sequences after two experiments were performed with an months. The order Clostridiales anaerobic granular sludge treating became the most important in agricultural wastes. The sludge was number, with Acetobacterium being incubated during two months at the most representative genus mesophilic temperature, with (49.8%). Archaeal diversity was less continuous CO injections in the important since almost all the headspace, creating an atmosphere of sequences were classified into the 100% CO. Liquid samples were same phylum, Euryarchaeota, for both collected after one and two months of the initial anaerobic microbial operation, for subsequent microbial consortium and the CO-adapted community analyses. Presence of population. After CO enrichments, the volatile fatty acids (VFA) and alcohols class Methanobacteria increased, with was verified by gas chromatography Methanobacterium becoming the most after two months. DNA was extracted representative archaeal genus (44.6% from the initial sludge and from the of the total archaeal sequences). On samples collected during enrichment the opposite, the class Red Text: Invited 59

Methanomicrobia, initially dominant, main fractions were obtained (juice decreased. The Ion Torrent and fibers). Washing of the fresh sequencing technology allowed us to biomass aims to reduce or remove the monitor the evolution of the microbial nonstructural ash (salt deposits). Both population during CO enrichments, fractions were tested for their total dry and gave insights on microbial species matter and ash content. Sugar particularly adapted to the monomer composition was tested for bioconversion of CO into butanol. both fractions applying acid hydrolysis Future work will now include using as described in (Sluiter et al., 2008a). known bacterial solvent producers to The extent of glucan-to-glucose bio-augment the adapted consortium, convertibility was tested for the juice to improve its butanol production and by enzymatic hydrolysis (by optimize and stabilize the performance simultaneous saccharification and of the process. fermentation (SSF)) and acid hydrolysis followed by fermentation by Ayah Alassali baker’s yeast. High Performance 1. Introduction: Abu Dhabi exemplifies Liquid Chromatography was used for a coastal desert, where seawater ethanol, sugars and other metabolites could be used for salt-tolerant crops analysis as described in (Sluiter et al., (halophytes) cultivation. The produced 2008b). halophyte biomass could be utilized in feed, food and/or energy production, 3. Results: The juice fractions were depending on its chemical found to represent 68.86 ± 1.78% of composition. In this study the UAE the unwashed batch and 74.09 ± native halophyte Salicornia sinus- 3.68% of the washed batch. The fiber persica was studied for its potential to fractions were found to contain be used as a feedstock for bioethanol 98.99% (unwashed biomass) and production. Fresh Salicornia sinus- 99.52 % DM (washed biomass) of perica contains more than 65% of which 19.77% (unwashed biomass) water. For such green biomass direct and 19.73% (washed biomass) was fractionation and fermentation can be ash. Dry matter content of the juices advantageous. This allows for water were found to be 13.06% (unwashed) preservation and the ability to run at and 11.58% (washed) of which lower dry matter in the fermentation 58.01% (unwashed) and 57.37 % step. Chemical characterization and (washed) was ash. Sugar analysis ethanol potential of the juice and revealed relatively low concentration fibers of the fractionated Salicornia of glucose, xylose, and arabinose in sinus-perica was examined in this the juice fractions (8.40 g/L glu, 5.97 study. g/L xyl, and 3.42 g/L ara in juice of unwashed biomass and 7.39 g/L glu 2. Methodology: Two batches of 4.87 g/L xyl, and 2.17 g/L ara in juice Salicornia sinus-persica (washed and of washed biomass) and not much unwashed) were juiced, where two difference was observed between the Red Text: Invited 60

washed and unwashed biomass. The Mike Guggenheimer, RSC Bio fiber fractions contained 15.6 g/100 g Solutions DM glu, 11.55 g/100 g DM xyl, and 14.04 g/100 g DM ara for the Abstracts unwashed biomass and 16.67 g/100 g DM glu 13.01 g/100 g DM xyl, and Benjamin Saydah 13.82 g/100 g DM ara for the washed Life cycle assessment (LCA) has been biomass. This is comparable to the used widely in recent years to lignocellulose content of the mature estimate the energy and (dry) plant (Cybulska et al., 2013). environmental implications of The highest ethanol yield in SSF with deploying algae-to-energy systems at Baker’s yeast was achieved in juice scale. Until recently, the emergent extracted from washed biomass after nature of the industry required that enzymatic hydrolysis. In this most LCA studies were based largely experiment 129.61% of the theoretical on assumptions about how the yield (based on sugar analysis) was industry would develop, likely achieved. This shows that fresh juice impacting the value of the results. For of Salicornia sinus-persica is a good this study data from a pilot-scale medium for yeast fermentation - but facility is used to estimate the life more work is needed to identify all cycle energy and emissions profiles of fermentable sugars in the juice. algae-to-biofuels production using hydrothermal liquefaction (HTL) as the Tuesday, December 10, 2013 | conversion pathway. HTL is attractive 4:00pm – 5:30pm because it is efficient, using the entire algae cell biomass rather than only Sustainability Performance the lipid fraction, and it is flexible, Value in Industrial producing a nonaqueous phase liquid product that can be blended during Biotechnology Processes petroleum refining and upgraded into

drop-in fuels, such as gasoline or Life Cycle Assessment of diesel. The pilot data is used to inform Transportation Fuel Production from the scale up to commercial scale and a Hydrothermal Liquefaction of Algae LCA is conducted on the estimates of Grown in Open Ponds commercial scale operation. The pilot Moderator: Benjamin Saydah, scale and commercial scale processes Sapphire Energy are then compared to current bio-fuels

process for both their Energy Return Is Wood Bioenergy Carbon Neutral? on Investment and Greenhouse Gas Roger Sedjo, Resources for the Future emissions. Results suggest that the

algal biofuel production based on HTL The Commercialization of Readily can provide substantial greenhouse Biodegradable, Biobased Fluids and gas (GHG) emission reductions Cleaners, and the Motivations Driving compared to petroleum-derived Their Adoption Red Text: Invited 61

benchmarks. Sensitivity analyses in anticipation of higher future reveal several bottlenecks currently demand. Empirical support for this holding back HTL-based algae-to- outcome is provided from the forest energy production including industrial experience of the U.S. since the mid- CO2, and nitrogen fertilizer supply 1990s. chains, biocrude yields and heat recycling. Efforts to address these in Mike Guggenheimer industry and the implications on The results of readily biodegradable, anticipated life cycle impacts are biobased fluids and cleaners can be discussed. seen across a wide range of industries and segments. In solely looking at performance, many readily Roger Sedjo biodegradable, biobased chemicals The view that wood biomass for have proven time and time again to energy is “carbon neutral” since its be high performers in comparison to use has approximately zero net traditional petroleum-based because carbon emissions released in counterparts. This single paper its utilization for bioenergy are submission would explore lessons subsequently captured in forest learned and successful regrowth has been challenged by the commercialization examples in a wide use of a static accounting view of the variety of industries (marine to utility forest carbon. The challenge argues fleets, waste segment and offshore oil that the use of wood for biofuels will & gas) seeing the benefits of result in a decrease in the forest stock renewable chemical solutions, such as and a net reduction in the carbon readily biodegradable, biobased captured in the forest, at least for an functional fluids and cleaners. These extended period. The argument of this products are developed from paper is the static approach ignores ingredients from a number of regular dynamic economic factors. The BIO meeting attendees. The single anticipated future use of forest for submission would also explore the energy provides incentives to increase commercial motivations (performance, forestry investments in forest stocks, value and sustainability) driving the which will increase carbon switch from conventional sequestration in the near term and petrochemicals to readily offset carbon losses due to increased biodegradable, biobased alternatives. harvesting of forests for energy. This situation is demonstrated with a Wednesday, December 11, dynamic optimization forest 2013 | 8:30am – 10:00am management model which shows that investments in the creation of forest Microbial Synthetic Pathways biomass can often anticipate future to Renewable Chemicals and uses and thus forest stock, and thus Biofuels sequestered carbon, can actually rise

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Bright Science at DSM: Development During the last 5-10 years a of a Commercial Lignocellulosic tremendous progress was made to Biomass-to-Ethanol Process continuously drive down Moderator: John Perkins, DSM manufacturing costs which posed major hurdles for commercialization. Screening and Analysis Pipeline for the It has been recognized that cost Microbial Production of Renewable optimization needs an integral process Chemicals and Fuels view (feedstock, pretreatment, Michael Leavell, Amyris enzymatic hydrolysis, yeast propagation and ethanol fermentation DEINOCHEM: The Deino Way to Green using high performing yeast) rather Chemicals than optimization of single Nagib Ward, Deinove SA technologies. This presentation will show how innovation and an Corbion: Developing a biobased integrated research approach are used products portfolio to drive yeast and enzyme Hans van der Pol, Corbion development programs. Latest results to improve thermo-tolerant enzyme Abstracts mixes, their production as well as advanced yeasts with improved John Perkins robustness and C5 sugar converting Royal DSM N.V. is a global science- properties will be discussed. based company active in health, nutrition and materials. DSM is a Michael Leavell frontrunner in applied industrial Amyris is an integrated renewable biotechnology to produce ingredients products company focused on for food and feed, or in bio-conversion providing sustainable alternatives to a technology for fuels or bio-based broad range of petroleum-sourced chemicals. products. Our industrial synthetic biology platform converts plant sugars Today’s market needs are driven by a into a variety of molecules that can be number of major global trends and used in a wide range of renewable challenges. At DSM we’re using our products, including ingredients in: innovative strengths to address some cosmetics, flavors and fragrances, of the most important of these trends polymers, industrial lubricants, and challenges, such as climate consumer products, and renewable change, energy independence. fuels. To enable rapid strain Advanced biofuels such as cellulosic development, a high-throughput strain ethanol offer excellent solutions to screening pipeline is needed as well as these challenges of today and even an in-depth analysis of winning strains more for future generations. using omics technologies. Together these technologies allow identification and characterization of improved Red Text: Invited 63

strains, enabling strain engineers to DEINOCHEM is focused on producing quickly iterate on strain designs. This renewable chemicals and reduced presentation will focus on Amyris’ high sugars from a variety of starting throughput screening platform and materials including cellulosic and other omics analysis tools, and present renewable feedstocks. The main costs specific examples of how each of biochemical production are technology enables strain concentrated on feedstock, enzymes development. and capital, with feedstock and enzymes often accounting to 50% or Nagib Ward more of the total cost. DEINOCHEM’s DEINOVE (Alternext Paris: ALDEI) is a platforms leverage thermophillic cleantech company that designs, organisms (45-55C) that are not develops and markets novel industrial subject to the common constraints or processes using biotransforming limitations often seen in industry, and platforms based on the untapped release their own enzymes in solution extraordinary and advantageous during the fermentation/hydrolysis, a properties of the Deinococci. Originally rare process advantage named founded in France, and principally Consolidated Bioprocessing (CBP). located in Montpellier, DEINOVE is Having the ability of produce end- now offering its technology platforms products and expressing the enzymes globally, with commercial necessary to break down feedstock for representation in both the EU and biotransformation, all at higher United States (www.deinove.com). temperatures, gives DEINOCHEM’s solutions a unique and significant cost Taking advantage of the unique advantage in techno-economic genetic properties and unusual analyses. robustness of this relatively unknown bacterial phylum, DEINOVE optimizes Due to the unique genetics of natural fermentation and metabolic Deinococci and essential towards capabilities of these bacterial "micro- biochemicals production, DEINOVE factories" to produce renewable green developed specific proprietary chemicals. The company is structured molecular biology tools to both tailor in two groups: while DEINOL is the end products production and focused on producing renewable optimize the metabolism of its advanced biofuels, DEINOCHEM is platforms. DEINOVE is the World focused on green chemicals or leading synthetic biology group intermediates. Of notice, two leveraging Deinococci for industrial subsidiaries operate independently, applications. These properties are key yet falling within the DEINOCHEM success factors for DEINOVE’s group: new antibiotics research targeted industrial applications by (DEINOBIOTICS SAS) and plastics allowing a platform customization for bioremediation (THANAPLASTTM). each commercial partner, independent

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of either the starting feedstock or relationships and a unique global pretreatment technology employed. supply chain. Sustainability is in the heart of our business. Based on the In summary, the exceptional strong foundation in technology, properties (robustness, biodiversity sustainability and cooperation, and metabolic properties) of Corbion is building a differentiated DEINOCHEM’s platforms, and biobased products portfolio. Thus, DEINOCHEM's developmental research Corbion products are designed by results, demonstrate the huge science, powered by nature, and potential for these microorganism’s to delivered through dedication. This become true bacterial micro-factories presentation will discuss the recent for the production of bio-based expansion of the Corbion products and chemicals in a cost-effective, project portfolio -into area's such as environmentally friendly way. succinic acid, FDCA and CaPropionate- and highlight key partnerships, Hans van der Pol capabilities and technologies that form Corbion (Purac) is built on a strong the basis for the development into a foundation of leading edge full-fledged biobased products fermentation and down-stream company. processing technologies, deep market understanding, strong customer

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