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 Steve Bobzin, Ceres, Inc. maximum highway weight limits per truckload. Abstracts

While Yulex built its business around John Fulcher guayule for rubber production, this Chromatin is a vertically integrated feedstock offers the greatest potential sorghum feedstock provider with for biorefineries to offer a wide array experience in biotechnology, seed, of products that come from mutually and feedstock growth, harvest, and complimentary processing delivery. Sorghum has seen technologies. In addition to its core resurgence in the US as a feedstock guayule rubber processing technology for advanced bioethanol with the platform, Yulex has been developing a added benefit of using the sorghum wide array of supporting and stover and/or high biomass sorghum complimentary guayule processing for the production of biogas. Sweet technologies that yield resins, waxes, sorghum is an attractive rotational oils, cork, energy (electricity and crop with sugar cane in Brazil. In biofuels), green building materials, addition, high biomass sorghum has and specialty chemicals. great potential for cellulosic in Brazil and the US. The growing need for Monday, December 9, 2013 | global non-food feedstocks over the 10:30am – 12:00pm next 20 years will be met by a combination of three primary sources: Advances in Reeds and Grasses energy crops; agricultural residues; and woody biomass. By 2030 it is Red Text: Invited 4

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, corn). Sorghum has a four-month high yields with favorable logistics growing season and in Brazil, sweet (extended harvest period, and good sorghum is being co-cultivated with bulk density). Undesired escaped Red Text: Invited 5

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 bench-scale test. Monday, December 9, 2013 | 2:30pm – 4:00pm In this presentation, recent achievements on the project will be Gases as Feedstock: The New summarized. Particularly, to reduce Renewable? Red Text: Invited 6

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 much lower cost than current fossil BioGTL platform for the conversion of fuels. These applications are natural gas to fuels and chemicals. particularly suitable for insular and Sol Resnick, Calysta Energy rural environments across the Pacific Rim. WRI'S CAT™ Technology for Carbon Capture and Re-Use Julia Allen Karen Wawrousek, Western Research First-generation biofuels dominate the Institute alternative fuels market today, but issues like indirect land use exchange Abstracts and food vs. fuel drive demand for new feedstocks. The glut of cheap Michael Schuppenhauer natural gas from shale as well as Current progress towards meeting trends toward the use of waste renewable advanced and cellulosic biomass feedstock open up biofuel goals are falling short of opportunities for new technologies expectations due to technical and that will alter the alternative fuels financial hurdles. However, those landscape. Overlapping technologies efforts have focused on a narrow set between gas-to-liquid (GTL) and of lignin containing feedstocks and the biomass-to-liquid (BTL) allow quest for liquid biofuels. Reviewing developers some degree of freedom to domestic and international large-scale choose the most attractive feedstock. yield data from alternative, novel BTL is inherently more capital- feedstocks, using a biochemical intensive than GTL due to the solids pathway (anaerobic digestion) to handling step. The future of BTL gaseous advanced biofuels would on a hinges on advancements in broad base offer a vast novel pool of gasification and pyrolysis technology. feedstocks and highly cost-competitive While BTL requires higher upfront production processes with 3-5 times capital, advantages include low cost of the yield per acres than pursuing certain feedstocks such as MSW as liquid biofuel routes, while being GHG well as the potential for long-term negative. In fact the electricity cost feedstock agreements. At current oil would be at 3-15 c/kWh and fuel cost prices, most thermochemical GTL and Red Text: Invited 7

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 Sol Resnick process, systems for biomass and Important progress has recently been water recycle, and a system for made toward engineering a number of biomass residue conversion to phototrophic and fermentative nutrients. The microorganisms used in microorganisms for biofuels WRI's CAT™ process show a rapid production. Several limitations, most growth rate, indicating efficient notably the ever-increasing cost and conversion of CO2 into biomass. linkage to oil prices of sugar However, unlike many biologically- feedstocks, currently prevent the based carbon capture and re-use economical production of biofuels from systems, WRI’s CAT™ process is not microbial systems. Exploiting dependent on light, which affords this methane, an inexpensive, domestically process multiple advantages. Not only abundant carbon feedstock, can the CAT™ technology be directly represents an attractive strategy integrated to an existing stationary towards economically sustainable CO2 source, it is able to operate 24 biofuel production. Calysta Energy has hours a day, year-round. Deployment developed a genetic engineering of this technology with large, platform for host organisms cylindrical reaction vessels reduces the (methanotrophs) capable of necessary land area to less than 2% metabolizing methane to a variety of of that required for equivalent biofuels and biochemicals. The genetic biodiesel production from open pond tools, together with innovative algae processes. Additionally, this fermentation and bioprocess closed system uses significantly less approaches, enable the rapid water than that required for algal implementation of well-characterized open ponds algae or renewable crops, pathways to utilize natural gas as a since evaporation is not an issue. biological feedstock instead of sugar. Economic analysis of the process predicts that the CAT™ process- Karen Wawrousek produced biodiesel will be WRI’s patent-pending CAT™ process is economically competitive with a novel biological carbon capture and petroleum-based diesel, and a re-use technology for the recycle of preliminary limited life cycle analysis carbon dioxide (CO2) from stationary of the CAT™ process estimates that Red Text: Invited 8

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 value of biofuels and biobased Making sugar from CO2 chemicals for industry partners. The Moderator: Kef Kasdin, Proterro company has developed a novel,

scalable biosynthentic process that Low-Cost Sugar from Energy Beets: A integrates a patented, highly largely Unexplored Opportunity productive microorganism with a Steve Libsack, Betaseed robust, modular photobioreactor made

from off-the-shelf materials. This American Process Cellulosic Sugar process yields a fermentation-ready Technologies “Sugar is the New sucrose stream, rather than a mixture Crude® in Asia” of sugars, allowing simple, low-cost Theodora Retsina, American Process, downstream processing. Proterro will Inc. present updates on it key

development milestones. The CO2 or Sugar to Vegetable Oil: company has scaled up its novel Commercial Scale Production of Pure photobioreactor design and conducted Vegetable Oil Through Plant independent functionality tests that Cell Cultures confirmed the unit’s robustness: the Karl Doenges, Sweetwater Energy photobioreactor is able to withstand

category 1 hurricane winds. Because Abstracts of the innovative photobioreactor

design and materials used, Proterro Kef Kasdin has also been able to validate low A critical barrier to scale for the fabrication costs. Proterro is in the production of biofuels and biobased process of commissioning a pilot plant chemicals is access to reliable and in Florida and has completed a economical feedstock. Feedstock is preliminary design, layout and the largest component of end-product associated cost estimate for a costs and is subject to the variables demonstration-scale plant. and challenges of various agricultural product markets, including sugarcane, Steve Libsack, co-author David corn and biomass. Current Bransby approaches to the cultivation, harvest, In North America, about 1.2 million storage, transport and processing of acres of sugarbeets are currently crop-based products add substantial grown for sugar production. These cost - when all that is required is Red Text: Invited 9

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 of simple sugars for the production of cellulosic feedstock. We have recently ethanol and many advanced conducted energy beet trials in the bioproducts. Energy beets will Southeast with good success. For typically produce twice the ethanol per example, an initial commercial-scale acre compared to corn, and may even test in Baldwin County, South compete with sugarcane. Specifically, Alabama, yielded 35 tons of beets per production costs are lower than for acre over the winter with no irrigation sugarcane, the pulp has much higher (http://blog.al.com/live/2013/05/man value (as animal feed) than sugarcane ufacturing_plant_cited_as_m.html). bagasse, the region where the crop This translates into approximately 830 can be grown is multiple times larger gallons of ethanol per acre, and at a than the region suited to production of price of $30 per ton for beets, a gross sugarcane, and it is possible that income for the grower of $1,050 per energy beets could supply sugar for a acre and a sugar cost of 10 - 15 cents longer period each year than per pound for the processor. Work on sugarcane. Therefore, energy beets irrigated beet production for have the ability to become a major conversion to ethanol is also underway industrial crop for all types of in California industrial sugars. In addition, they can (http://www.ethanolproducer.com/arti be used as high energy, high value cles/9978/lowering-ethanols-carbon- livestock feed as whole beets, or beet footprint-with-energy-beets). Our pulp: in the Southeast, beet pulp panel will provide an overview of the retails for over $700 per ton when potential of energy beets for sold in 50-lb bags as horse feed. production of biofuels and Achieving the 2007 federal mandate bioproducts, including the agronomy (Energy Independence and Security of the crop and possible end-uses. Act) to produce 36 billion gallons of renewable fuels each year by 2022, Theodora Retsina will depend on crops like energy At American Process, we believe that beets. Being a simple sugar plant, it Sugar is the New Crude®. Our vision can easily be utilized to produce is to be the leading biomass pre- industrial sugars without the use of treatment technology in the world, enzymes and the need to first convert with a value proposition that combines Red Text: Invited 10

engineering capabilities, quality of capital efficient manner. Other technical support, assurance of high applications include pellet-making yields, clean sugars, and minimum facilities and biomass power plants. total cost of operation. We have Our first large-scale implementation of developed two proprietary biorefinery Green Power+ technology is a technologies for producing low-cost biorefinery pilot plant in Alpena, cellulosic sugars from non-food based Michigan, capable of converting about biomass. 20 tons/day of hemicelluloses to sugars and co-products. The plant The Green Power+® technology is a capacity is up to 2 million gallon/year patented technology for the of ethanol. The Alpena biorefinery production of low-cost cellulosic started up in Q3/2012. The AVAP sugars from the hemicelluloses of process produces low-cost C5 and C6 biomass. sugars from both cellulose and hemicellulose of biomass feedstocks. Our AVAP® technology is a patented The AVAP process employs a solvent technology for the production of low- for lignin and sulfur dioxide to cleanly cost cellulosic sugars from the fractionate biomass into cellulose, cellulose and hemicelluloses of any hemicellulose, and lignin. Optionally, biomass. the cellulose may be recovered as a cellulose material or precursor for The two technologies address different making cellulose derivatives. The market opportunities for biorefineries cellulose that is produced by the AVAP in the Asia Pacific region and process is extremely susceptible to worldwide. For each technology, any hydrolysis by enzymes to produce biomass may be utilized, including glucose, thus greatly reducing enzyme hardwoods, softwoods, and costs. We see AVAP as having agricultural residues. The Green tremendous potential in Asia in areas Power+ process produces low-cost C5 of abundant, competitively priced and C6 sugars from the hemicelluloses biomass feedstock. An advantage of of biomass feedstocks. Sugars are this technology is that it can operate extracted from the solids which are with softwood, hardwood or then utilized for existing applications, agricultural residue, thus increasing in synergy with biomass-based the number of locations where it can renewable power, pellets, sugarcane be deployed. The cellulose sugars plants, and many other existing sites. from the AVAP process can be Co-location minimizes capital costs for converted into a variety of high-value commercial implementation. We see chemicals, many of which are high tremendous opportunity for Green demand in Asia either on their own or Power+ configurations Asian sugar as a feedstock for plastics cane ethanol facilities, where our manufacture. Our first large-scale technology can substantially increase implementation of AVAP technology is the production of ethanol in a very a biorefinery pilot plant located in Red Text: Invited 11

Thomaston, Georgia. The Thomaston Renewable Oil Feedstocks for biorefinery is capable of converting the Pacific Rim about 10 tons/day of biomass to sugars, ethanol, and co-products. The Moderator: Sanjay Wagle, AliphaJet plant capacity is up to 300,000 gallon/year of ethanol or other Naveen Sikka, TerViva products, such as butanol, jet fuel, or biochemicals. The Thomaston Kirk Haney, SGB biorefinery started up in the first half of 2013. Mike Cey, Ag-West Bio Inc

Karl Doenges Abstract The recent volatility of commodity Speakers will discuss how they have prices and rapid pace of industry begun to commercially establish their development is stimulating the feedstocks, with specific emphasis on: biorefining and oleochemical industry (1) Business models -- contemplated to diversify its raw material feedstock, structures, process for validation, and using an array of fats and oils. The ultimate winning models with ability to take various fats and oils is a customers (2) Prioritization -- how the key risk mitigation strategy; however, feedstock company prioritized reengineering the way oils are made landowner customer segments, addresses the feedstock problem at a geographies, and downstream market more fundamental and sustainable opportunities (3) Focus on Asia Pac -- level. This presentation will review evaluation of feedstock Sweetwater Energy’s deployment of a appropriateness for Asia Pacific new CO2 or sugar to pure vegetable countries. oil technology, whereby a modular, CO2 source-flexible, commercial scale Wednesday, December 11, oil production technology has been 2013 | 8:30am – 10:00am optimized for the biodiesel, renewable diesel and oleochemical Oilseed Derived Feedstocks - industries. This talk will highlight: 1) The basic science behind CO2 and Can they compete with sugars to pure vegetable oils, 2) using Petroleum? this technology as a bolt-on for corn ethanol plants and the resulting Moderator: Jack Grushcow, Linnaeus synergies, and 3) characterization of Plant Sciences Inc. the vegetable oil to enable optimum downstream processing Jonathan Burbaum, U.S. Department of Energy Tuesday, December 10, 2013 | 4:00pm – 5:30pm Andy Shafer, Elevance Renewable Sciences, Inc.

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evaluating the potential impact of David C. Bressler, Biorefining ‘low-cost’ petrochemical plants now Conversions Network being built in China. This discussion will examine how multiple companies are looking to highlight the value of Abstract their bio-based offerings, to have successful market entry and growth Oilseed Derived Feedstocks - Can they despite the low-cost challenge. compete with Petroleum? Andy Shafer Elevance Renewable Sciences and Renewable Chemical Wilmar International Limited have Platforms begun commercial shipment of specialty chemicals from their World- Monday, December 9, 2013 | Scale biochemical refinery in Asia. The 8:30am – 10:00am biorefinery, which has a capacity of 180 KTPY (400m lbs.) and is Differentiation for Bio-based expandable to 360 ktpy, is Elevance's first biorefinery using its proprietary Chemicals metathesis technology. With commercial capacity now available, 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- Bando Takehiko, Mitsubishi Chemical performance chemicals in end user applications. This paper will discuss Commercialization - Elevance's World the status and progress of the Scale Biorefinery has commenced commercialization activity taking place production and New Products have and the benefits the bio-based entered the Market - An update on technology is bringing to the progress and what's coming downstream markets. Andy Shafer, Elevance Renewable Sciences, Inc. Monday, December 9, 2013 |

10:30am – 12:00pm Abstracts

Damien Perriman Expanding the Oleochemical Jeffrey Hsu Value Chain to Bio-based Bando Takehiko Consumer Products In addition to all the other challenges Moderator: Tom Beardslee, Verdezyne of commercializing bio-based chemicals, Asian markets are Hirzun Mohd Yusof, Sime Darby Red Text: Invited 13

Biosynthetic Technologies (BT) holds Sam Bhargava, Jarden Applied exclusive patented technology that Materials synthesizes plant and animal oils into high performance synthetic oils used Biobased Synthetic Chemistry in lubricants, industrial chemicals and Allen Barbieri, Biosynthetic personal care. These “biosynthetic” Technologies base oils meet or exceed the performance characteristics of existing Abstracts petroleum based synthetic oils. Having Tom Beardslee tested BTs biosynthetic oils Hirzun Mohd Yusof extensively, several of the world’s Sam Bhargava largest manufacturers of automotive The conventional processing of crude and industrial lubricants are certifying vegetable oils produces an array of finished products they will bring to molecules in the current oleochemical market under their brand names. The value chain such as fatty acids, fatty chief formulator of a leading global alcohols, and dimer acids that are motor oil company recently stated used in the manufacture of consumer that these are “the most exciting products ranging from surfactants and products to enter the lubricants lubricants to polymers and cosmetics. industry since the introduction of PAOs These oleochemicals provide a [synthetic lubricants] 50 years ago.” renewable alternative to First-mover advantage, lack of viable petrochemical-based products. New competition and patented technology technologies are being developed that make BT a market leader in the broaden the reach of oleochemicals sustainable chemicals sector. into new products and are providing bio-based alternatives for consumers. Business model: BT is a wholesale This panel assembles speakers manufacturer/distributor of biobased representing multiple points along the synthetic oils that can be used by value chain of converting oleochemical finished motor oil, lubricant and inputs into bio-based consumer chemicals products manufacturers. products. From plantation and BT is currently working with dozens of vegetable oil refining to renewable leading global lubricant and additive chemical conversion technology to manufacturers as well as automotive polymerization and product and equipment manufacturers who are applications, the presentations show each testing, formulating and the current value chain as well as the preparing to launch motor oils and future potential value chain of the lubricant products based on BT’s oleochemical industry. biosynthetic oils.

Allen Barbieri Manufacturing: BT contracted with OVERVIEW Albemarle to build a demo manufacturing plant which is currently Red Text: Invited 14

operating in Baton Rouge. In late fall, life, lower maintenance costs and 2013, BT will begin construction of a increased fuel efficiency. full scale continuous flow production plant. Water pollution: BTs biodegradable oils could significantly reduce the ENVIRONMENTAL BENEFITS environmental impact of improperly dumped motor oil and oil runoff from Sustainable: BTs oils are made from roads, which accounts for 40% of fatty acids found in plant oils including water pollution in the U.S. (EPA). In canola, soy, palm, and coconut, as fact the amount of used motor oil and well as refined animal fat (all 100% lubricants that enter the world’s renewable carbon). oceans is the equivalent of one Exxon Valdez spills every week. Biodegradable: Using the most stringent testing protocols, BTs oils all Monday, December 9, 2013 | exceed the industry standard of 60% 2:30pm– 4:00pm biodegradation within 28 days. Renewable Chemicals and Non-Toxic: BTs oils tested non-toxic, Consumer Products even at contamination levels 100 times higher than the level needed to Renewable Chemicals: The Path to claim non-toxicity. Commercialization

Moderator: Max Senechal, Metabolix Not bioaccumulative: BT’s products do not bio-accumulate in living DEINOL: The Deino Way to Biofuels. organisms. Emmanuel Petiot, Deinove SA

Greenhouse gas reduction: A Life Leveraging a Consumer Brand for B2B Cycle Analysis of Greenhouse Gas Pull - The Nexa Project Emissions (GHG) from BT’s products is Jeff Uhrig, Bioformix 83% lower than the GHG emissions associated with poly-alpha olefin Chemo-catalytic conversion of (PAO), a product of similar function cellulose into para-xylene and use. John Bissell, Micromidas

Machine life / fuel efficiency: Abstracts Numerous tests have shown that BTs biosynthetic oils keep metal surfaces Max Senechal cleaner and facilitate lower friction and Metabolix, Inc. is an advanced scarring on bearing surfaces than the biomaterials company that is well highest quality synthetic oils. All of positioned to address growing market this keeps leads to longer machine demand for sustainable solutions in the plastics, chemicals and energy Red Text: Invited 15

industries. The Metabolix scientific In this presentation, Metabolix’s vice foundation and core science is the president, biobased chemicals, Max metabolic pathways for the production Senechal, will outline the Company’s of a class of microbial biopolymers – strategy for bringing these renewable polyhydroxyalkanoates (PHAs) – from chemicals to market and review renewable resources. PHAs can be various strategic options available to tailored through metabolic pathway biobased chemical producers in engineering and have a wide range of commercializing their innovative applications in industry, including use products. Key question addressed: Is in polymer form as biobased plastics the “drop-in” chemicals strategy or in monomer form as chemical currently pursued by several intermediates. The first commercial participants in the space the only product using the fermentation route approach worth considering for are biopolymers marketed under the successful market entry? Mirel and Mvera brands. Emmanuel Petiot Metabolix is also focused on DEINOVE (Alternext Paris: ALDEI) is a developing biobased four-carbon (C4) cleantech company that designs, and three-carbon (C3) chemicals. The develops and markets novel industrial C4 program is currently at the processes using biotransforming semiworks scale (80,000L) and platforms based on the untapped Metabolix has shipped samples of extraordinary and advantageous biobased gamma butyrolactone (GBL) properties of the Deinococci. Originally to prospective customers for testing, founded in France, and principally and GBL has been successfully located in Montpellier, DEINOVE is converted to BDO via hydrogenation. now offering its technology platforms Metabolix is making steady progress globally, with commercial with its C3 program as well – the representation in both the EU and Company is currently running United States (www.deinove.com). fermentation at the 20L scale and recently successfully recovered acrylic Taking advantage of the unique acid from biomass using its genetic properties and unusual proprietary FAST process. In 2013, robustness of this relatively unknown Metabolix is continuing fermentation bacterial phylum, DEINOVE optimizes scale up, engineering of microbial natural fermentation and metabolic strains, and development and capabilities of these bacterial "micro- optimization of its FAST recovery factories" to produce renewable green technology to produce biobased C4 chemicals. The company is structured and C3 chemicals to match chemical in two groups: while DEINOL is industry specifications for quality and focused on producing renewable purity. advanced biofuels, DEINOCHEM is focused on green chemicals or intermediates. Of notice, two Red Text: Invited 16

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 generally concentrated on feedstock, efficiencies and reduced raw material enzymes and capital, with feedstock and variable costs. and biotransformation accounting generally at least 50% of the total In summary, the exceptional cost. However, most processes are properties (robustness, biodiversity constrained to operate at the optimal and metabolic properties) of DEINOL’s temperatures of yeast or commonly platforms, and DEINOL's used bacteria (~35C), not maximizing developmental research results, on biotransformation kinetics, favoring demonstrate the huge potential for infections and preventing from using these microorganism’s to become true enzymes at their optimum activity bacterial micro-factories for the levels. production of biofuels in a cost- effective, environmentally friendly DEINOL’s platform leverage way. thermophillic organisms (45-55C) that are not subject to these constraints or Jeff Uhrig limitations, and release their own While B2B sales channels are enzymes in solution during the attractive to resource limited start- fermentation/hydrolysis, a rare ups, the consumer channel presents a process advantage named nearer term commercial opportunity, Consolidated Bioprocessing (CBP). allowing companies to build brand Having the ability of produce end- equity, demonstrate market adoption products and expressing the enzymes and de-risk larger scale B2B necessary to break down feedstock for applications. This presentation will metabolism, all at higher address how Bioformix created a temperatures, gives DEINOL’s consumer brand - Nexabond 2500 - solutions a unique and significant cost and a respective in internet and Red Text: Invited 17

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, other aromatic species. The including the ketal-based JAVELINTM presentation will discuss (1) the Technology, is positioned for specific advantages of the chemical significant global adoption with pathway, (2) which feedstocks have renewable alternatives that cut been tested and will be scaled first, greenhouse gas emissions and (3) the comparative projected address toxicity issues in everyday economics of the process, and (4) the items. Segetis has successfully performance of our recently developed and commercialized constructed pilot plant. specialty plasticizers for flexible PVC used in everyday objects including Tuesday, December 10, 2013 | flooring, shoes and packaging, as well 2:00pm– 3:30pm as formulation aids used in consumer and industrial markets such as Processes Leading to detergents and hard surface cleaners. Commercialization Levulinic Acid, the starting point for

these innovations, is the Making it Real: Levulinic Acid as a decomposition product of sugars and Biobased Building Block of a carbohydrates, the most abundant Renewable Chemical Industry feedstocks on Earth. Levulinic Acid is Moderator: Atul Thakrar, Segetis readily transformed into a wide variety

of chemicals and has the potential to Strategic Commercialization of Bio- become a foundational biobased based Chemicals chemical building block of a renewable Sue Hager, Myriant Corporation chemical industry. Despite high levels

of interest, the lack of large-scale, Developing new platform molecules: a cost effective, clean Levulinic Acid case study Red Text: Invited 18

production has hampered the growth Marcel Lubben of levulinic-based products. Segetis Royal DSM N.V. is a global science- has made a breakthrough in making based company active in health, Levulinic Acid a reality with a nutrition and materials. DSM is a disruptive process that enables frontrunner in creating bio-based and industrial-scale Levulinic Acid environmentally sound solutions, and production using a wide array of seeks to demonstrate the commercial biomass feedstocks, including viability of renewable technologies in fructose, glucose and cellulosic sugars. collaboration with strategic partners in The combination of our ability to make the value chain. The development and cost effective Levulinic Acid at scale supply of high value knowledge, with the market development for ingredients and expertise in the field JavelinTM plasticizers and formulation of bio-conversion technology are aids positions Segetis to meet the critical success factors determining its increasing demand for high future. DSM’s strategy is to license its performance, healthier, biobased technology and expertise to bio-based materials. entrepreneurs, enabling them to convert biomass in the most Sue Hager commercial and sustainable ways The chemicals industry is experiencing possible. Today’s market needs are a fundamental shift as cost- driven by a number of major global competitive bio-based chemicals trends and challenges. At DSM we are become a commercial reality. Leading using our innovative strengths to this transformation is the market address some of the most important availability of bio-succinic acid, a of these trends and challenges focused platform molecule used as a direct on supporting the transition from replacement for petroleum-based fossil to renewable raw materials. The succinic acid and also as a investment wave in the bio-based fundamental building block chemical in economy started from the early 00s. the production of numerous industrial After almost one decade, tremendous and consumer applications. Myriant is and impressive progress has been one of the world leaders made in technology development for commercializing bio-succinic acid with renewable chemicals and materials. two operating plants in production. Pioneers in the field are making their Myriant’s presentation will discuss the first commercial steps. company’s pathway for strategic financing of its flagship commercial In this speech we shall present a case plant, its commercialization strategy study to illustrate our vision on what for the production of bio-succinic acid, drives success in the as well as its plans for accelerated commercialization of bio-based development of its bio-based chemicals and materials: partnerships chemicals pipeline. & capital, feedstock choice, excellent operations, application know-how and Red Text: Invited 19

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. Satish Khanna, Provivi,Inc

Ajay Parikh, ZCL Chemicals Jayant Sawant Anand Ghosalkar, Praj Corporation Butadiene (BD) is a top 10 petrochemical primarily produced by Biocatalytic Preparation of heavy crackers. With more US Cyclopropanes – A Novel and Cost- crackers shifting to shale gas based Effective Approach light crackers, a net drop in production Moderator: Pedro Coelho, Provivi, Inc of butadiene has created a demand- supply imbalance and sharp price Abstracts swings in global markets.

Alternatively, butadiene can be Satish Khanna produced from renewable feedstocks Ajay Parikh such as sugars using greener routes Anand Ghosalkar via 2,3 butanediol-fermentation and In recent years, biocatalysis has subsequent conversion to BD by established itself as a key technology catalysis. Several research groups, for the production of fine chemicals. start-ups and few petrochemical The fine chemicals industry utilizes majors have shifted focus to bio- this technology to catalyze chemical butadiene. The focus of our study is to reactions with unprecedented regio develop deeper insights into the and enantioselectivity. technical and market driven Implementation of biocatalysis challenges of bio-BD production and therefore often results in a decreased shortlist solutions to overcome the number of chemical steps required for challenges. We present a detailed the synthesis of a given fine chemical, techno-economic analysis of bio- and ultimately reduces the cost of butadiene production starting with manufacture. In addition, biocatalysis sugar based BDO fermentation and favors mild conditions, resulting in purification by solvent extraction decreased waste streams, decreased followed by catalytic conversion of energy input, and improved BDO to BD. Detailed sensitivity safety. With an ever expanding Red Text: Invited 20

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 Our initial product platform is based on breakthrough technology for Jeff Fornero, Glucan Biorenewables cyclopropane biosynthesis developed at and licensed from the California Abstract Institute of Technology. There, we developed the most active catalyst CBiRC is developing renewable ever reported for olefin chemical platforms from bio-based cyclopropanation, a key reaction in the innovations and new ventures utilizing synthesis of drugs, hormones, and tools, components and materials being insecticides (Coelho et al., Science explored by the Center. Core 339, 307 (2013); Coelho, et al. Nat. knowhow and technologies include Chem. Biol. 9, 485 (2013)). The bioengineering of fatty acid and combined annual global market size polyketide biochemistry in for products that are accessible via microorganisms, as well as an Provivi’s technology exceeds $7 innovative and complimentary billion. Active pharmaceutical portfolio of developments in chemical ingredients (APIs) that contain a catalysis. By combining biocatalysis cyclopropane motif have a combined and chemical catalysis CBiRC creates market of over $5 billion. In addition, new knowhow and powerful pyrethroid insecticides containing a technologies that have the potential to cyclopropane motif have a market size nurture a sustainable bio-based of about $2 billion. Here we will briefly chemical industry. CBiRC believes the describe the technology, and then existing petrochemical supply chain focus on product applications and can be transformed with key market opportunities pursued by foundational intermediates that deliver Provivi. an array of drop-in chemistry or similar functionality to existing fossil- Wednesday, December 11, carbon-based chemicals. Here we will 2013 | 8:30am – 10:00am describe our progress towards Red Text: Invited 21

creating an advanced manufacturing start up of our Hugoton, Kansas system with new platform molecules commercial scale cellulosic facility in for biobased chemicals. December 2013. Abengoa also started operations at a demonstration scale Advanced Biofuels and municipal solid waste to ethanol facility in Spain, and is negotiating a Biorefinery Platforms commercial scale MSW facility today. We are also pursuing efforts to Monday, December 9, 2013 | produce other advanced biofuels and bioproducts from the same agricultural 8:30am – 10:00am residue and MSW feedstocks. We

currently are sourcing and managing Cellulosic Biofuels: Unleashing 380,000 tons of ag residues annually Commercial Production for our Kansas facility, and have already conducted 4 harvests. We also Moderator: Brian Conroy, BP are planning to utilize dedicated energy crops at some point in the near The Reality of Commercial Scale future. These efforts, along with start Cellulosic Ethanol Production up of the Ineos facility and progress in Christopher Standlee, Abengoa development and construction by Bioenergy US Holding, Inc. other producers of Advanced Biofuels make second generation biofuels a Meeting worldwide demand with reality. cellulose Steve Hartig, POET-DSM Advanced Steve Hartig Biofuels Cellulosic bio-ethanol is poised to play a crucial role in the energy landscape Cellulosic ethanol: Scale-up and of the United States, driven by recent commercialization process and technology gains as well Markus Rarbach, Clariant as a commitment by government leaders to prioritize domestic, Delivering Big by Getting Small sustainable energy production. But Wes Bolsen, Cool Planet Biofuels worldwide, the potential is even greater. In the U.S., liquid fuels Abstracts demand has fallen in recent years, and most experts predict this trend Christopher Standlee will continue. However, worldwide Abengoa is an international leader in demand, driven largely by growth in the development of Advanced Asia, is expected to increase Biofuels. We have pursued a 10 year dramatically in coming decades. The effort to produce cellulosic ethanol potential for expansion of cellulosic from multiple agricultural residues, bio-ethanol in those areas is culminating in the commissioning and enormous. China, as the second- Red Text: Invited 22

largest corn producer in the world, is constitutes an almost carbon neutral uniquely suited to reap the rewards of new energy source using an already commercialization efforts such as existing renewable feedstock that those by POET-DSM Advanced doesn’t compete with food or feed Biofuels. China has stated that it will production and land use. The field of not use corn for ethanol production; application is wide, from second however, the abundance of crop generation biofuel to the chemical residue that remains after harvest industry. A key controversial issue does provide ample feedstock for regarding technological developments biomass-based ethanol production. aimed at the production of cellulosic The Energy Information Administration ethanol is the commercial economic expects China to lead the world in viability of the process. In order to be demand growth for liquid fuel by competitive on the fuel market, the 2035. India is expected to experience production costs must be comparable the next-largest demand growth, and with the production costs of the rest of Asia is not far behind. manufacturing conventional bioethanol Cellulosic bio-ethanol can help meet from plants containing sugar or that demand. POET-DSM Advanced starch. The challenges facing process Biofuels is constructing a 25 million- development therefore include gallon-per-year cellulosic bio-ethanol optimization of the ethanol yield while plant in Emmetsburg, Iowa that will lowering operational and capital costs use corn cobs, leaves, husk and some such as the reduction in enzyme costs stalk as feedstock. Farmers this fall and energy efficiency improvements. will harvest an estimated 120 Clariant’s sunliquid® technology offers thousand tons of biomass to feed this an efficient and economic process for commercial biorefinery. Construction the production of cellulosic ethanol. It is on schedule for startup in early overcomes the main challenges of 2014. POET-DSM is now working to competitive conversion of license this technology to other lignocellulosic feedstock into cellulosic ethanol producers, and we see Asia as sugars for fermentation to cellulosic an important opportunity for putting ethanol. In July 2012 a demonstration this technology to good use. Sound plant with an annual output of 1000 technology, created here in the U.S., tons of ethanol started operation. This is ready to provide solutions for is the last step on the way to energy demands well into the future. commercializing a technology platform for second generation biofuels and Markus Rarbach biobased chemicals. The plant Cellulosic ethanol made from represents the complete production agricultural residues has been a chain, including pretreatment, scientific and commercial interest for process-integrated production of decades however the development feedstock and process specific and commercial deployment of enzymes, hydrolysis, simultaneous C5 technologies have been limited. It and C6 fermentation and energy Red Text: Invited 23

saving ethanol separation. Thus, a sustainability in transport, energy high process yield of 20-25% can be independence and create green jobs achieved and cellulosic ethanol and income for the agricultural sector. production becomes competitive to first generation ethanol. The process Wes Bolsen itself is energy neutral, yielding Carbon negative fuels is a reality cellulosic ethanol with about 95% of today through the combination of CO2 emission reductions. However, biofuels and biochar. Wes will talk the process is flexible for use of about the company’s ability to different feedstock and different commercialize Cool Planet’s production plant concepts. After technology through a much smaller, successful operation of over four years distributed plant model. He will lay of pilot plant operations, on 20 July out transformative industry models 2012, the sunliquid demonstration such as mainframe computers plant was officially commissioned in becoming personal computers, and the Lower Bavarian town of Straubing. the mass production of micro-refinery The plant replicates the entire process technology paralleling other chain on an industrial scale, from pre- industries. He will finish by discussing treatment to ethanol purification, their first commercial facility and how serving to verify the viability of with the backing of Google, GE, sunliquid technology on an industrial ConocoPhillips, BP, and their other scale. On an annual basis, up to 1,000 marquee investors, they were able to tons of cellulosic ethanol can be commercialize. produced at this plant, using approximately 4,500 tons of wheat Monday, December 9| 10:30am straw. In May 2013, first runs with – 12:00pm corn stover and sugarcane residues also showed good results. This was a Production of Drop-In new milestone reached by the project, Hydrocarbon Fuels from confirming that the technology is Cellulosic Biomass flexible for different feedstocks and can be implemented worldwide. The Moderator: Thomas Foust, National results obtained in the demonstration Renewable Energy Laboratory plant are being incorporated into plans for the first production plant. The Charles Cai, University of California, worldwide potential for cellulosic Riverside ethanol is huge, in the transport sector as well as the chemical Jesse Q. Bond, Syracuse University industry. Liquid energy sources will continue to play an important part as DME, its advantages and its promise energy source for transportation. through small-scale production Thus, cellulosic ethanol can make a Brittany Syz, Oberon Fuels huge contribution towards more

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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 particularly desirable for heavy-duty considerations will be discussed, and vehicles and air travel. Furthermore, perspectives will be offered on leading enzyme costs for biological conversion options and challenges. of lower cost cellulosic biomass to ethanol tend to be high. Catalytic Next, Jesse Bond from Syracuse routes are being developed to convert University will summarize pathways biomass sugars and their breakdown for catalytic processing of these fuel products such as furfural, 5- precursors and identify appropriate hydroxymethylfurfural (HMF), and catalysts for each fuel option. In levulinic acid that we call fuel addition, theoretical yield limits and precursors into hydrocarbon fuels and current performance will be described fuel components compatible with the for the various fuel options, and existing fuel infrastructure. A potential opportunities for improvements will be advantage of this aqueous processing outlined. Tom Foust from the National approach to such “drop-in” fuels is Renewable Energy Laboratory (NREL) that only thermochemical conversion will then provide an overview of the Red Text: Invited 25

National Advanced Biofuels DME has storage and handling Consortium (NABC) and describe advantages over other diesel NABC partnerships to accelerate alternatives, such as CNG and LNG. In commercial production of hydrocarbon current demonstrations using bioDME fuels. Included will be an outline of in Europe, Volvo has shown 95% key metrics that must be met for reduction in CO2 emissions with commercial viability and the stage of companies such as DHL using DME in development of various processes to commercial operations. See meet these goals. http://www.biodme.eu

The range of topics covered in this Oberon Fuels has developed an panel will allow the audience to better innovative, small-scale production understand the pathways, progress, method that produces DME in regional and challenges in converting cellulosic fuel markets, bypassing the initial biomass into fuels and fuel need for a national infrastructure. In components that are compatible with these regional fuel markets, regional the existing aircraft and heavy and feedstocks are used to produce DME light duty vehicle infrastructures. to be utilized in regional trucking hauls, creating new regional Brittany Syz economies and green jobs. The first DME (dimethyl ether) is a clean- such production facility will be online burning, non-toxic, diesel alternative. in Imperial Valley, in Brawley, CA in Its high cetane value and quiet August 2013. The regional model combustion, as well as its inexpensive supports not only local job growth and propane-like fueling system, make it clean air initiatives, it also provides a an excellent, inexpensive diesel viable economic structure that allows alternative that will meet strict for capital expenditure in sync with emissions standards. market growth – it really hits the triple bottom line concept of DME has been used for decades as an sustainability. energy source in China, Japan, Korea, Egypt, and Brazil, and it can be The need: produced domestically from a variety of feedstocks, including biogas (plant In his climate initiatives speech in and food scraps) and natural gas. July, President Obama identified that Ideal uses in North America are in the we need to address heavy-duty transportation, agriculture, and trucking. Why? Because 30% of the construction industries. Because particulate matter in our atmosphere production is not dependent upon the is caused by commercial trucking. price of crude oil, DME will have stable DME produced using the Oberon Fuels pricing, which is competitive with that method is actually carbon negative – of diesel. because it uses feedstocks that would otherwise release methane into the Red Text: Invited 26

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 Decision and Risk Analysis in performance level of any of the diesel Industrial Biotechnology alternatives. Alan Propp, Merrick & Company

On June 20th, Mack Trucks also Project Financing Considerations announced they plan to commercialize John M. May, Stern Brothers & Co. DME trucks. In its press release, Mack

Trucks noted that DME “offers many Critical Questions for Bolt-On environmental and societal benefits, Biorefineries: Avoiding Unintended including that it can be made from Consequences multiple sustainable feedstocks,” Doug Dudgeon, Harris Group Inc. including grass clippings, animal waste and other sustainable sources. Abstracts

See:http://www.aboutdme.org/aboutd Alejandro Zamorano Cadavid me/files/ccLibraryFiles/Filename/0000 The Renewable Fuel Standard 00002405/Release_Mack_2013-06- celebrated its seven-year anniversary 20.pdf on 1 May 2013. Bloomberg New Red Text: Invited 27

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 business case to attract needed bolt-on application, in order to avoid funding. This presentation will unintended consequences from describe the Decision and Risk integrating new technology with an Analysis process and present an existing, revenue-generating asset. example of how the science was These questions concern the applied to an industrial biotechnology compatibility of core technologies, project. economic trade-offs between the existing process and the new process, John M. May infrastructure availability, and the Project Finance Fundamentals 1. commercial affinity of the host’s Current State of the Bank Market 2. current business with the potential Typical Project Finance Structure 3. new business. Drawing on experience Project Structure Mitigates Project from Harris Group’s decades of Risk 4. Project Capitalization 5. experience in advanced biofuels and Successful Financing Requires retrofit plant design, this paper Systematic Approach 6. At Financial examines the critical questions of bolt- Close Project Finance Execution 1. ons for implementing advanced Sources and Uses 2. Private biofuels technologies in three Red Text: Invited 28

contexts: a dry-grind ethanol plant, a the physiochemical parameters sugarcane mill and ethanol plant, and resulted in about 14.3 g/l of butanol in a pulp mill. 120 h using acid treated jatropha seed cake hydrolysate (7% w/v) along with Tuesday, December 10, 2013 | AnS components. The process was 2:00pm – 3:30pm scaled up to 15 L level yielding 18.6 g/L of butanol in 72 h. This is the first Progress in Production and report wherein high yield of butanol Commercialization of Alcohol has been obtained in single batch fermentation using acid pretreated Fuels jatropha seed cake. The strain was found to be tolerant to 3.0 % butanol Moderator: James J. Spaeth, U.S. under optimized conditions and the Department of Energy tolerance could also be demonstrated by the strain’s ability to accumulate Production of Biobutanol from rhodamine 6G. The strain was found Jatropha Seed Cake to be having high expression of the Jasmine Isar, Reliance Industries stress-response protein GroEL, the Limited reason behind the tolerance and growth of the strain in presence of 3.0 Clean technology innovation to % (v/v) butanol in the medium. produce advanced biofuels, reduce landfilling and simulate regional Sam Park economies The commercialization of advanced Sam Park, Enerkem biofuels is driven by the need to reduce dependency on oil, increase Advances in DOE supported Cellulosic energy diversity and reduce carbon Ethanol Projects footprint. Public policies around the James J. Spaeth, U.S. Department of world contribute to the expansion of Energy biofuel production by stimulating both the supply and the demand for Kevin Gray, Chemtex renewable fuels. Despite the global recession, the advanced biofuels Abstracts industry now operates facilities, and a first wave of technologies is now ready Jasmine Isar to be commercialized, as large scale Butanol production by Clostridium facilities currently being built. A wide acetobutylicum ATCC 4259 was variety of biomass and residues are studied using the renewable feedstock used as feedstock for these projects, jatropha seed cake,. Chemical including municipal solid waste. Three mutagenesis for improvisation of the main categories of technologies are strain for better butanol tolerance and being developed and commercialized production was done. Optimization of in this sector. The ones capable of Red Text: Invited 29

using municipal solid waste (MSW) as Kevin Gray a feedstock provide an alternative to landfilling and incineration, thereby Tuesday, December 10, 2013 | offering additional environmental 4:00pm – 5:30pm benefits. MSW feedstock has a large potential, given its abundance, and Overcoming Challenges in the fact that it is already collected. In Regulation and Intellectual addition to providing a new source of Property energy, advanced biofuels facilities stimulate regional economies, create Regulatory Strategies for Use of high quality jobs, provide synergies Genetically Modified Organisms in with traditional sectors such as Biofuel Production forestry, agriculture and waste, and Moderator: David J. Glass, D. Glass contribute to revitalize the industrial Associates, Inc. manufacturing sector.

Kevin Wenger, Mascoma Corporation The presentation will talk about the context for the growth of this sector What is patentable in the biotech and for the use of MSW as a feedstock sector? for the production of advanced Konrad Sechley, Gowling Lafleur biofuels. It will describe the main Henderson, LLP categories of technologies that were developed over the last decade and The Here and Now of TSCA give an example of a technology using Regulations on Biobased Chemicals non-recyclable municipal solid waste Kathleen Roberts, Bergeson & as a feedstock. The benefits of Campbell Consortia Management, LLC commercial-scale advanced biofuels facilities, as well as the advantages of Abstracts using municipal solid waste will also be presented. Finally, the presentation David J. Glass will explain why public policies around Kevin Wenger the world play an important role in Genetically modified microorganisms stimulating innovation and creating a are broadly coming into use for the business climate that attracts private production of biofuels or bio-based investment to allow for the chemicals. These include modified commercialization of these yeast strains with improved or more innovations. efficient ability to ferment ethanol,

photosynthetic microorganisms to James J. Spaeth produce fuels from solar energy, and DOE will provide an update on microbes that can synthesize alkane progress and developments in DOE fuels or fuel precursors. These are in supported cellulosic ethanol projects. addition to the numerous industrial

enzymes used in ethanol Red Text: Invited 30

fermentations that are now produced Next, Kevin Wenger will discuss in modified production strains. Many Mascoma’s experience with its 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 Red Text: Invited 31

protect biotech related inventions? knowledge, commercial success will be Much uncertainty exists in the virtually unachievable. 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. Abstract Kathleen Roberts How do the evolving biobased and Eric Mathur renewable chemical technologies fit Drop-in replacements for petroleum within the current Toxic Substances liquid fuels represent a major Control Act (TSCA) framework? The component of the growing energy short answer is “not well.” The longer demand. Although cellulosic ethanol answer involves detailed evaluation, will mitigate the depletion of oil probable notification, reporting, and reserves used for gasoline and algal recordkeeping requirements. This biofuels hold long term promise as jet session will focus on helping fuel substitutes, there are no obvious manufacturers understand the journey present day renewable feedstocks their biobased product will face in the available. Candidates include canola, U.S. Environmental Protection oil palm, rapeseed and soybean; yet Agency’s current system of TSCA redirecting agricultural resources regulation, including potential delays, toward energy feedstocks negatively detours, and pitfalls. We will consider impacts food security and is not strategies, tips and helpful hints that sustainable. A promising alternative is companies can use as they navigate the non-edible oilseed shrub, Jatropha the TSCA gauntlet. Without this curcas. Jatropha is native to Central Red Text: Invited 32

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 maximized time for accumulation of Jatropha from becoming the preferred selected and unselected mutations. oilseed feedstock for renewable jet Jatropha planting materials were fuel. collected throughout Central America in order to create a germplasm Karl Seck repository which now comprises over Red Text: Invited 33

Mercurius Biorefiningwas established Synthetic Biology, to produce profitable drop-in fuel (Jet Fuel and Diesel) from non-food Algae and Marine biomass feedstocks. Mercurius is Biotechnology developing a “faster ~cheaper~ better” method of producing profitable Monday, December 9, 2013 | drop-in biofuels. Mercurius uses a 8:30am – 10:00am proprietary, patent pending technology called Renewable Acid- hydrolysis Condensation Hydrotreating Optimizing Algae Production for (“REACH”) to make profitable drop-in Beneficial Bioproducts hydrocarbon liquid fuels. Microalgal-based Food and Feed Jay Long Ingredients – Crop of the Future? This presentation describes strategies, Moderator: Jim Flatt, Synthetic Genomics motivations, and future plans for integrating aviation biofuel into normal Marine Microalgae for a Sustainable airline operations. As aviation biofuel Future-Production and Application of technologies evolve, driving production Algae for Fuel, Feed and rates up and unit costs down, the use Nutraceuticals of alternative aviation fuels as part of Valerie Harmon, Cellana LLC the standard fuel sourcing operation is drawing closer to a reality. Alaska Algae, beyond energy Airlines flew 75 flights using an 80/20 Jose Sanchez, OriginOil blend of HEFA based bio-jet in 2011 as part of a “proof of concept” initiative Commercialization of New Algae-based aimed to support the aviation biofuel Omega-3 Supplements with High production efforts. The airline is Bioavailability currently engaged in a project with Brian L. Goodall, Valicor Renewables Hawaii BioEnergy for a project to LLC produce aviation bio-fuel in an area that currently sees very high jet fuel Abstracts prices and significant supply constraints. It is Alaska’s opinion that Jim Flatt identifying these types of markets will Projected growth in world population not only allow the producers to and changes in dietary trends, coupled compete on an economic basis, but will with constraints on arable land, water also provide much needed supply and carbon emissions will increasingly integrity. tax conventional agricultural food production methods and drive development of alternative crops which can augment traditional sources of food and feed products. Large- Red Text: Invited 34

scale cultivation of microalgae has the too expensive and limited in scale to potential to meet a significant portion compete on price and availability of future needs, however, requires today. However, advances in significant development to establish synthetic biology and genomics offer cost-effective and robust production of powerful new tools to improve strain strains with desirable nutritional performance and robustness, which properties. Microalgae are relatively are required for attractive production unique in their ability to produce a economics. Strains exhibiting large fraction of their biomass as high- improved photosynthetic efficiency quality protein as well as other relative to wildtype ancestor strains nutrients including oils and carotenoid are required to achieve higher pigments. Researchers at Synthetic biomass productivity and titer Genomics have assembled a vast and associated with economical production 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 Red Text: Invited 35

catalog of algal strains for mass since often times the same amount, if cultivation. To date, Cellana has not more energy is expended screened a large collection of algal generating algae than is generated by strains and produced more than 10 it. Current methods for separating metric tons of biomass from numerous algae from water such as centrifuge natural marine algal strains for the and membranes are simply too development of biofuels, feed, and expensive for large-scale high-value nutraceuticals. Cellana’s implementation. Centrifuge in research and production at KDF have particular can also damage the algae addressed major areas that are crucial product rendering it useless in many for the commercialization of algal applications. Enter OriginOil, biofuels: yield improvement, cost developer of a chemical-free, high- reduction, and the overall economics. speed technology that efficiently 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 Red Text: Invited 36

burgeoning market applications for benefits of EPA (such as anti- high quality algae. Using the recent depression) it is pure EPA, rather than OriginOil findings regarding shelf life an EPA / DHA mix, that is important and astaxanthin production, Riggs will for effectiveness. In addition, the EPA educate attendees about new growth dose taken is less critical than how sectors in the algae industry. much of the EPA ends up in the blood stream and target organs – the Brian L. Goodall, co-authors Isaac bioavailability of the EPA source. In Berzin, Sam Couture, Kiran Kadam & the market today, there are products Brian Waibel Qualitas that are more bioavailable, like Krill Health and Valicor Renewables have oil, which is premium priced and partnered to commercialize the first enjoys market growth of 40% per algae-based pure EPA supplement. annum. It is important to remember 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 Red Text: Invited 37

Tim Hsiau, University of California, Berkeley James Carothers Naturally-occurring metabolic systems Ginkgo Bioworks: A factory for have evolved control circuitry that engineering organisms minimizes the accumulation of toxic Jason Kelly, Ginkgo BioWorks intermediates and maintains homeostasis. At present, engineering Abstracts even rudimentary control circuitry for synthetic biological systems remains Vivek Mutalik very difficult. We have successfully Our ability to routinely engineer formulated design-driven approaches genetic networks for applications is that use mechanistic modeling and limited by the scarcity of a well kinetic RNA folding simulations to 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 Red Text: Invited 38

Cheap and reliable gene synthesis Metabolic Engineering of methods will benefit the field of Aquatic Photosynthetic biotechnology. To address this Organisms problem, we have developed an inexpensive gene synthesis and error Moderator: Peter Heifetz, Heifetz correction method. In our system, we BioConsulting can make hundreds of genes in a one- pot fashion from microarray- Mark Hildebrand, University of synthesized oligos. We then California, San Diego developed a genetic system for selection of correct sequences from Margaret McCormick, Matrix Genetics, the background pool of error- Inc. containing genes. Our genetic system can also provide information about the David Stern, Cornell University solubility of the synthesized gene. Such solubility information will be Farzad Haerizadeh, Life Technologies useful in ranking algorithms for rational design of genetic and Abstract metabolic pathways. Aquatic photosynthetic organisms, Jason Kelly including cyanobacteria, micro algae, Organism engineering firms are diatoms and macro algae (seaweeds) emerging that build microbes to have attracted significant interest as specification for customers. Making potential sources of heavy usage of robotics and photosynthetically-derived and computer-aided design (CAD), these carbon-neutral renewable feedstocks, companies are closer to specialty chemicals and biofuels. The semiconductor chip fabrication plants application of metabolic engineering than to biotech labs. Ginkgo tools and approaches to cyanobacteria Bioworks opened a new 11,000 sq ft and algae creates opportunities for organism fabrication facility in coupling bioproduction to direct solar early 2012. This talk will describe the energy conversion without the need facility and our work for organic carbon feedstocks. Recent designing and building microbes for advances in the understanding of the production of chemicals and biological circuitry and its regulation in fuels -- including our development of these organisms has now enabled organisms engineered to use C1 effective application of technologies compounds as feedstocks. for strain selection, targeted genetic manipulation and synthetic biology to Monday, December 9, 2013 | chemicals, food, feed and medicines. 2:30pm – 4:00pm In parallel, product-optimized cultivation and product isolation strategies have evolved rapidly as Red Text: Invited 39

projects have moved toward ethanol and biodiesel) and industrial commercialization. This session will chemical production. However, highlight significant advances in selection of microalgal strains with metabolic engineering of desirable physiological and cyanobacteria / algae as well as the development of integrated and biochemical features from local scalable platforms for specialized habitats has been one of major bioproduction. challenges in the large-scale production of microalgal biofuels and Tuesday, December 10, 2013 | chemicals. Although the coastal areas 2:00pm – 3:30pm of Pearl River Delta have long been known for huge diversity of aquatic Updates on Algal Biofuel life surprisingly only little work has Production in China been done to use local microalgae resources for biofuel and chemical Production of Biofuels and Chemicals production. Recently, we have Using Microalgae Isolated from the Coastal Regions of China isolated 18 marine microalgal strains, Moderator: David Guangyi Wang, belonging to the families Tianjin University Chlorellaceae, Scotiellocystoidaceae, Neochloridaceae, Selenastraceae and Development of Jet Biofuels for a Scenedesmaceae. Of isolated strains Sustainable China Aviation Industry Mychonastes afer PKUAC 9 and Pengcheng Fu, Beijing University of Chemical Technology Scenedesmus abundans PKUAC 12 were selected for biomass and ethanol Synthetic photosynthetic production analysis. Comparison of Cyanobacteria to produce fine three cultivation modes (stationary, chemicals directly from CO2 shaken and aerated) resulted in the Weiwen Zhang, Tianjin University highest biomass productivity obtained

for aerated cultures that yielded 0.09 Efficient full use of Algae biomass g and 0.11 g dry weight per day per components for production of liquid liter of medium for M. afer PKUAC 9 fuels and S. abundans PKUAC 12, Guanyi Chen, Tianjin University respectively. Carbohydrate Abstract accumulation monitored by FTIR showed that early stationary phase is David Guangyi Wang optimal for biomass harvest. Microalgal biomass has been Microalgal biomass was successfully considered as one of the most used as a carbohydrate feedstock for promising feedstocks for biofuel (e.g., fermentative bioethanol production. Red Text: Invited 40

Scenedesmus abundans PKUAC 12 from traditional biofuel development. was superior feedstock for bioethanol Jet bifuels will incorporate second- production when pre-treated with the generation methodologies relative to sustainable feedstock source selection combination of dilute acid treatment and fuel processing, which are and cellulase. Additionally, two uniquely suited for aerospace strains, Chlorella Sorokiniana 21 and applications. These jet biofuels can Monoraphidium 3s35, were found to potentially be blended with traditional produce large amount of long-straight kerosene fuel (Jet-A) to reduce chain alkane and fatty alcohols under dependency on petroleum-based fuels. the optimized cultivation conditions Additionally, sustainable feedstock sources may avoid deforestation 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 and food industries. dioxide outputs. Details will be 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 Guanyi Chen demand/distribution for aviation biofuels; 2) demonstration flights and Rapid growth of Economy in China 3) Supply-chain R&D focused on algae promotes the booming urbanization feedstock and processing techniques. and automobile use thereby leading to the increasing oil demand. Future oil The study may result in the source is considered to be coming environmentally progressive fuel from biomass, particular algae solutions (bio-jet fuels) that differ biomass, through efficient conversion Red Text: Invited 41

process. Currently algal biomass may An industrial biotechnology pipeline for be converted into liquid fuel optimizing isoprenoid production in substituting for petroleum oil by Saccharomyces cerevisiae extracting its oil components followed Moderator: Sunil Chandran, Amyris by esterification process. In this case, non-oily components may be not well Systematic level engineering E. coli used. This paper is addressing this and S. cerevisiae efficiently producing bottleneck and presents research work aromatic amino acid and ethanol done in our research group. Liu Shuangping, Jiangnan University

To this end, our technical route Renewable Chemicals from Waste consists of the following steps：1）30- Carbon 40wt.% oily component are extracted Lisa Dyson, Kiverdi followed by esterification process to produce biodiesel; 2) non-oily Clarification of succinic acid component is subjected to fermentation broth by ultrafiltration hydrothermal process to produce and the associated membrane fouling crude bio-oils; 3) bio-oils are mechanism upgraded further into high-quality of Jianmin Xing, Institute of Process liquid fuels. In this case, all Algal Engineering, CAS biomass components are converted to liquid fuels. Abstracts

The results obtained indicated that the Sunil Chandran yield of liquid fuels produced from Amyris is building an integrated algae, which is suitable for vehicle renewable products company to apply use, may be up to 60-70wt.%, much industrial synthetic biology to higher than biodiesel yield from only genetically modify microorganisms, oily component, while the production primarily yeast, to serve as living cost may be reduced by 75-80% per kg of oil. The market penetration of factories. These modified yeast strains such liquid fuel seems a little bit more convert plant-sourced sugars into promising and competitive. The potentially thousands of isoprenoid research may be quite valuable in molecules, providing a broad range of promoting algae oil development in renewable chemicals and the coming years. transportation fuels. Synthetic biology

approaches at Amyris have Tuesday, December 10, 2013 | 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 Red Text: Invited 42

pipeline and its application for decreasing the acetic acid formation; combinatorial exploration of thousands genetic switch on or off the expression of complex genotypes to continuously of target genes to conciliate cell improve isoprenoid-producing strains growth and precursors synthesis; over the past five years. attenuate the shunting pathway to construct non-growing cells; and also Liu Shuangping, co-authors Liang increase the enflux by expression the Zhang, Guiyang Shi exporter. After these works, an National Engineering Laboratory for industrial E. coli with high productivity Cereal Fermentation Technology and yield was constructed which was (NELCFT) is an advanced biomass comparable or even better than biotransformation research center in existing producing strains. Based on China. Some microorganisms the chassis above, the organisms producing organic acids and ethanol producing D-phenylalanine, L- are being building by applying phenylglycine and tryptophan are industrial synthetic biology in NELCFT. being constructed. Besides, our team In microorganisms, the control also constructed a super S. cerevisiae architectures of the biosynthesis at the system level with the abilities of pathway for desired chemicals always higher permeability resistance, lower 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 synthetic biology, the costs and time bio-based replacements to are decreased to develop an effective oleochemicals and petrochemicals cell factory at systematic level. The using our proprietary Carbon biosynthesis of aromatic amino acids Engineering Platform, a bioprocessing (like phenylalanine, tyrosine and technology capable of converting low- 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 pathways. In this study, the fraction of the cost of current engineering of a L-phenylalanine (Phe) approaches. Kiverdi’s technology uses producer was carried at systematic a class of proprietary level: modify the PTS system to chemoautotrophic microbes that act moderate the glucose uptake rate as “whole cell” biocatalysts in our innovative 1-step bioprocess for the Red Text: Invited 43

direct production of renewable organic 10kDa) were used and two models molecules from gaseous inorganic were applied to analyze the fouling carbon and hydrogen for a wide mechanism. Results indicate that variety of product applications such as ultrafiltration is feasible in clarifying surfactants, polymers and fuel succinic acid fermentation broth. additives. Almost all the microorganism cells (99.6%) were removed from the With increasing levels of waste fermentation broth. Proteins were also generation and decreasing available removed effectively by all membranes space for disposal, each year 134M selected in this study. The removal tons of landfill, 170M tons of rate was 79.86% for PES 100kDa, agriculture and 60M tons of forest 86.43% for PES 30kDa, 86.83% for waste generated in the United States PES 10kDa, and 80.06% for the RC is an untapped biomass resource to 10kDa. After ultrafiltration, clearer produce high value chemicals. permeate was obtained comparing Kiverdi’s solution fills a “scale gap,” with the centrifugation. Among all that enables localized, low CapEx tested membrane, PES 10kDa gave deployment of waste-derived syngas the best results which showed relative conversion plants to target diverse higher initial flux (18.86 L/m2 h), chemistries to make drop-in and highest protein removal rate (87%), custom renewable chemicals with mid and less flux drop rate. Resistance-in- to long carbon chain lengths, varying series model was applied to determine 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 Red Text: Invited 44

most membranes increased after patented and third party verified ultrafiltration. LipiTriggerTM technology to produce high value algae products. T2e’s Wednesday, December 11, LipiTriggerTM and grow tank 2013 | 8:30am – 10:00am technology not only increases algae biomass productivity and lipid content Techno-economic and Physical but also improves algae lipid profiles Analysis for Microalgae Growth favorable for biofuels production. T2e's novel intellectual

property uses a highly sustainable, Moderator: Mary Rosenthal, Algae waste resources-based Biomass Organization mixotrophic approach in conjunction

with our patent pending grow tank Algal Lipid Trigger and Grow Tank system to provide certain strains of System algae with the most optimal controlled Mark Randall, T2e Energy, LLC growing environment. This approach

has achieved unequalled results Techno-economic and Fluid Dynamics including greater algal density while Analysis for Growing Microalgae with favorably changing the oil profile that the Intent of Producing Biofuel Using a serves the biochemical/bioplastics and System Model biofuels industry. Controlling every Leah Raffaeli, University of Denver variable of the algal growth process

enables T2eH to produce a predictable A Dynamic Imaging Cell Monitoring level of oil and biomass production System for Real-Time Analysis of with a symbiotic modular system co- Algae-to-Biofuel Production located near existing coal, gas, or Victoria Kurtz, Fluid Imaging wood fired power plants. With this Technologies, Inc. approach T2e can produce a gallon of

algal oil at scale for less than Algae Testbed Public Private $1.00/gallon anywhere in the world. Partnership (ATP3): An Experimental Framework for Performing Long Term The presentation will cover results Cultivation Trails Across Different from UCSD’s Center for Algae Regional, Seasonal, Environmental, Biotechnology, US Department of and Operational Conditions Agriculture and Ohio State’s Ohio John McGowen, Arizona State BioProducts Innovation Center that University, AzCATI verify the level of algal growth including total biomass, lipids and the Abstracts resulting change in oil profile. We will also discuss our unique methodology Mark Randall for our mixotrophic system that T2e Energy Holdings ("T2e") is a start- includes our patented algae grow tank up alga to biofuels company located in system that enables T2e to grow algae San Diego dedicated to use its Red Text: Invited 45

adjacent to existing power plants energy inputs, carbon emissions, and anywhere in the world. environmental sustainability. No algal growth scenarios included in this study Leah Raffaeli come close to meeting the U.S. Techno-economic and systems studies requirement of biofuel emitting at to date are abbreviated and missing a least 20% less carbon emissions than number of important variables. By diesel from crude oil. including these variables in a detailed model integrating biology, chemistry, Victoria Kurtz engineering, and financial aspects, a This presentation will detail a novel more defined fluid dynamics and new system (patent applied) for real- financial analysis are possible. time analysis and monitoring of algae Through optimizing the model production. The system uses in-flow productivity based on the resulting net digital imaging to capture images of profit, the system analysis results in a all representative cells or other more accurate assessment of microorganisms in photo bioreactors environmental and economic or raceway ponds. Sophisticated sustainability of specific algal growth image processing algorithms are used scenarios. Photobioreactor algal in real time to segment each growth scenario optimization in the microorganism from the background, system model has resulted in realistic and record over 30 size, shape and engineering design requirements. gray-scale measurements for each Results show feasibility for microorganism. Cell size and 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, Red Text: Invited 46

including how the particle images are university and commercial facilities in acquired and measurements made. Hawaii (Cellana), California (Cal Poly Finally the results of the analysis will San Luis Obispo), Ohio (Touchstone be shown, detailing how the system Research Laboratories), and Georgia can be used to monitor microorganism (Georgia Institute of Technology), size and concentration, and in ATP3 provides comprehensive particular, identification of predators. cultivation and harvesting facilities along with the required experience of John McGowen operating an open testbed system. ATP3 is made possible by a $15 million U.S. Department of Energy A primary objective is to utilize our competitive grant from its Bioenergy team’s expertise and world-class Technologies Office. Our vision is to facilities to perform long term establish a sustainable network of cultivation trials with process and regional testbeds that empowers statistical rigor, producing robust, knowledge creation and dissemination meaningful datasets across different within the algal research community, regional, seasonal environmental and accelerates innovation, and supports operational conditions. These data are growth of the nascent algal fuels critically important to support TEA and industry. Our goal is to create a LCA activities that will guide research network of operating testbeds, and development towards the bringing together world-class transformative goal of cost- scientists, engineers and business competitive algal biofuels by 2022. To 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 Red Text: Invited 47

testbed sites via the adoption of a Computational Design: Enabling New uniform design of outdoor raceway Products, New Enzymes, and New pond systems with respect to: size, Pathways geometry; volume; depth and Eric Althoff, Arzeda Corp. hydrodynamic mixing. Identical outdoor raceway ponds have been Developing novel strategies on laccase installed at each site using a standard production and purification: Recent geometry, paddle wheel design, and advances and new trends monitoring and control systems. As it Feng Wang, Institute of Process is also essential that each site Engineering, CAS operates to a standard set of protocols with respect to experimental study An Industrial Scale Platform for timing and duration, operating Enzymes and Other Proteins conditions, as well as sampling and Danai Brooks, Dyadic International, analytical protocols for a fixed set of Inc. parameters, we have spent Phase 1 harmonizing protocols across all sites. Abstracts Without these controls for our cultivation systems and protocols, the Tim Hitchman inter-site variability would be more Degumming is the removal of difficult if not impossible to interpret. phospholipid impurities from oil. This For this presentation, we will present step results in the majority of loss of current progress on the initial oil yield associated with purification of experimental design framework, our crude soybean and canola oils, among harmonization and alignment activities others. The reason for the yield loss is and results from our initial cultivation that phospholipids are emulsifiers and trials. drag oil with them as they are removed. Enzymatic degumming with Technical and Research Purifine® PLC is a unique process that results in increased yield of Presentations degummed oil by reducing heavy phase oil losses and release of the Monday, December 9, 2013 | DAG component of the gums. The 8:30am – 10:00am value of implementing Purifine PLC degumming is further enhanced by Development of New Industrial integration into downstream refining Enzymes (for edible oil or biodiesel end uses) and improvements in quality of co- products such as meal. Enzymes for The biotech (r)evolution in oilseed degumming of vegetable oils are crushing & refining hardly a new thing, having first been Moderator: Tim Hitchman, DSM introduced to the

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industry in the early 80’s. Since then, techniques, we have recently a steady stream of technical developed a new algorithm, called publications has demonstrated that Enzyme Identification™, to rapidly degumming using enzymes is feasible engineer enzymes with known and can be applied to deliver oil that catalytic mechanisms for non-native exceeds standard quality substrates. For a given chemical specifications. Only in recent years, reaction, our algorithm screens “in the economic benefits of enzyme silico” large databases of structural degumming have also been quantified, and sequence information to allow the those processors who have taken the rapid discovery of existing enzymes lead in adopting the technology for that possess both the necessary degumming of catalytic machinery and an crude soybean oil are now appropriate substrate binding pocket. documenting yield gains of up to 2%. For those enzymes that possess the These data now allow the industry to necessary catalytic machinery but lack establish benchmarks for overall an active site that can accommodate economics of investment for future the substrate(s) of interest, Enzyme implementations. This practical case Identification automatically redesigns about technology developments in the and remodels the active site pocket oilseed crushing & refining industry thus enabling catalysis of the desired will illustrate how to fast forward reaction. The demonstrated success evolution into revolution and provide and wide applicability of our methods direction for successful biotech open the way for the design of a commercialization. variety of novel biocatalyst necessary for the efficient development of 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, Red Text: Invited 49

biopolymer modification and induced mutation and continuously biosensors. However, low enzyme bioengineered since. Addresses the yield in laccase production and high- critical bottlenecks of protein cost in laccase purification limit its use discovery, development, scale-up and in industrial applications. Great efforts commercialization. Enables new have been made to enhance laccase product introduction with less time, production, involving isolation and cost and risk. Enables new product breeding of high-producing strains, introduction with less time, cost and medium optimization, inducers risk. Broad platform capabilities utilization, reactor design and validated through 17 years of R&D recombinant expression of laccase and 15 years of product sales and genes in different hosts. Methods for partnerships with key players. laccase purification have been developed for different microbial Monday, December 9, 2013 | species, including ammonium sulphate 10:30am – 12:00pm precipitation, ultrafiltration, three- phase partitioning, gel filtration, ion Stabilization in Fermentation exchange, and affinity Processes chromatography. It is crucial to improve laccase production and New Yeasts for Grain Based Ethanol purification for the high demand of Moderator: William Kenealy, Mascoma laccase from industry. In this paper, Corporation we critically review recent advances in laccase production and evaluate the Study on anaerobic fermentation of advantages, drawbacks, and different organic wastes for biogas scalability of developing technologies production for laccase purification. In addition, Priyanka Gupta, Birla Institute of novel strategies for the enhancement Technology, Mesra, Ranchi of laccase yield and magnetic separation methods for simple and Production of ethanol from pine efficient purification of laccase are put needles using different fermentation forward according to our recent strategies achievement on laccase research. Parvez Singh Slathia, Shri Mata

Vaishno Devi University Danai Brooks

What is C1? A robust and versatile Continuous Fermentation: Prospects fungal platform for gene discovery, for Future Rollout expression and the production of Joshua Velson, Nexant enzymes and other proteins. Based on the Myceliophthora thermophila Abstracts fungus, a soil-borne saprophyte that secretes cellulases. Developed in the William Kenealy early 1990’s through a fortuitous UV- Red Text: Invited 50

Mascoma has developed advanced process of anaerobic digestion thereby yeast products for grain-based ethanol generating biogas, which can be used production. By expressing starch- as renewable energy source. Biogas is degrading enzymes in situ during the a source of energy which will leave fermentation, TransFerm™, can less of an environmental footprint, as reduce the cost of separately compared to coal or oil. The objective purchased enzymes as well as of this study was to see the increase the yield and rate of ethanol performance of the selected organic production. The genetic modifications wastes in a single stage anaerobic used to create TransFerm™, are built reactor for biogas production. In this in a robust ethanol production strain, study, experiments were conducted resulting in a yeast product that is using different organic wastes i.e. truly a “drop-in” solution for ethanol vegetable waste, garden waste, saw producers. Transferm™ Yield + is an dust, fruit wastes, and sugarcane improved strain that also makes less bagasse, under anaerobic conditions. glycerol thereby resulting in a higher Mine water (from Jitpur colliery, India) yield compared to conventional yeast. was used as inoculum for the biogas The increased yield was enabled by production. The study was carried out introduction of an alternative in a specially designed laboratory metabolic pathway designed to scale reactor for a period of two complement the reduction in glycerol. months. Amongst the considered The in situ production of glucoamylase biomasses the highest methane yield also reduces the concentration of of 212.87cc was observed with glucose present initially during vegetable waste. 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 Red Text: Invited 51

In the current study we have tried to combined gave better sugar yield use pine needles from Pinus roxburghii when enzymes were used in isolation. as a source for ethanol production. In fermentation studies, it was seen Pines are coniferous, evergreen, that SSF proved to be a better method resinous tree belonging to the genus yielding more ethanol concentration Pinus of the family Pinaceae. Pine than SHF. While using biomass loading needles from Pinus roxburghii were of 5g in the pretreatment processes obtained from forests of Tikri, fermentation yield of .2g of ethanol Udhampur, J&K (India). The Pine was obtained as the best production. needles were dried and comminuted Further, Kulyveromyces marxianus to paas through a sieve of 2 mm. was a better producer of ethanol Different concentrations of acid having a higher conversion efficiency (H2SO4)and alkali (NaOH, KOH, of 52% than that of Saccharomyces aqueous NH3)were used in cerevisiae which is 47%. The biomass conjunction with steam and pressure residue left after first pretreatment for varying time intervals for has considerable amount of sugar maximizing the sugar release. The which was used for ethanol biomass stream after pretreatment concentration. Thus, pine needles are was filtered and pH adjustment was an attractive source for ethanol made in the filtrate. The solid fraction production and technology needs to be was dried and again treated with developed for their use in fuel ethanol steam for obtaining sugar from this industry. fraction. Enzyme treatment of both the fractions was done using Joshua Velson 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 Red Text: Invited 52

the future. Designs including free-cell way to produce non-fossil hydrogen chemostats, immobilized cell (H2) through high temperature (750– bioreactors and membrane-integrated 1500°C) conversion of carbonaceous bioreactors will be discussed. material into a synthesis gas (syngas) mainly composed of carbon monoxide Monday, December 9, 2013 | (CO), carbon dioxide (CO2) and H2. 2:30pm – 4:00pm The final calorific value of syngas, that is its H2 to CO ratio, varies depending Advancements in Biological on the feedstock type used and Processes in Industrial gasification conditions.

Biotechnology One typical way of achieving

augmentation of the H content of Moderator: Bhima Vijayendran, 2 syngas in industry is through the use Redwood Innovation Partners, LLC of the water-gas shift (WGS) reaction:

CO + H2O → CO2 + H2. The WGS is Performance of a Carboxydothermus v catalyzed using catalysts that are hydrogenoformans gas lift reactor for highly intolerant to sulfur and need syngas upgrading into hydrogen regeneration, and the process is Mathieu Haddad, University of energy-intensive, namely for steam Montreal generation.

Biohydrogen Production from Alternatively, biologically-mediated Lignocellulosic Biomass WGS reactions possess several Chunzhao Liu, Institute of Process advantages over conventional Engineering, CAS chemically mediated WGS

mechanisms; such as enzyme Thermophilic Consolidated specificity, improved yields, overall Bioprocessing of Cellulosic Materials to cost efficiency and environmental Biohydrogen attractiveness. Lew Christopher, South Dakota School of Mines and Technology-CBRD Of interest to this study is the obligate

anaerobic Carboxydothermus Planetary Sustainability and Biological hydrogenoformans, an extreme Resource Utilization in Space thermophilic (70°C) bacillus that uses John Cumbers, SynBioBeta CO as sole source of carbon and

energy and catalyses the WGS

reaction. Abstracts

We investigated and optimized, in a Mathieu Haddad, co-author Serge R. 35 L gas-lift reactor, H conversion of Guiota 2 CO by a pure culture of C.

hydrogenoformans. The reactor was Gasification of biomass and waste is a Red Text: Invited 53

operated with a continuous supply of disrupt recalcitrant structure of gas for 3 months. Reactor lignocellulosic biomass for improving performance was evaluated under hydrogen production. Achievements in various operational conditions, such our research group together with gas recirculation (0.3 and 1.5 L/min), recent progress around the world will CO loading rate (from 0.05 to 0.46 be critically reviewed in this paper, -1 –1 mol.L reactor.d ) and nutrient addition and future perspectives in the field of to the medium. Overall, results biohydrogen production from indicated a constant H2 yield of lignocellulosic waste will be outlined. –1 95±1% and 82±1% (molH2·mol CO) in An emphasis is given on an efficient a supported and unsupported growth hydrogen production process via medium respectively regardless of the thermophilic fermentation of cornstalk operational condition tested. Once the by co-cultures of Clostridium biomass concentration was sufficiently thermocellum and C. dense, a maximum CO conversion thermosaccharolyticum integrated –1 -1 activity of 0.17 molCO.L reactor.day or with alkaline hydrolysis. Together -1 3.79 LCO/Lreactor.day was achieved. these results will establish the most One major parameter that impacted important conditions to explore for both biological activity and volumetric future process development on mass transfer was the gas biohydrogen production. recirculation/CO feed ratio. We here demonstrate that, as long as this ratio Lew Christopher is higher than 40, mass transfer An environmentally friendly and limitations are bypassed resulting in potentially viable alternative for maximum conversion efficiency sustainable H2 production is presented (90.4±0.3%) and biological activity through the utilization of renewable –1 (2.7±0.4 molCO.g volatile suspended solid cellulosic materials. Switchgrass –1 (VSS).day ). (SWG) is viewed as one of the most promising energy crops for the U.S. Chunzhao Liu conditions with low nutrient/water Lignocellulosic biomass is considered requirements and high adaptability to as one of promising resources for the any weather/soil/land conditions. production of renewable energy. The Production of SWG is projected to microbial conversion of the increase ten-fold within next ten lignocellulosic biomass into hydrogen years. Furthermore, its high is attracting increasing interest carbohydrate (>65%) and low lignin because hydrogen is as one of the (<20%) content favors a fermentation most promising energy carriers for the route for SWG utilization. On the other future. However, this bioconversion hand, the handling and disposal of efficiency is still limited because of low municipal solid waste (MSW) is of biodegradability of the lignocellulosic growing global concern. In the U.S. biomass. Therefore, there is a need to only, MSW recently reached 5 lb per develop an efficient technique to capita per day. However, MSW Red Text: Invited 54

contains approximately 60% of Novel Solid Catalysts for Production of biodegradable material which can be Renewable Sugars and Chemicals from utilized for bioenergy production. The Lignocellulosic Materials H2 production capabilities of the Moderator: Brian M. Baynes, Midori extreme thermophile Renewables, Inc. Caldicellulosiruptor saccharolyticus DSM 8903 were examined on SWG Lignocellulosic biomass as potential and MSW, with glucose and substrates for the white biotechnology microcrystalline cellulose (MCC) Joachim Venus, Leibniz Institute serving as references. It was Agricultural Engineering (ATB) demonstrated that C. saccharolyticus can ferment SWG bioH2 in a single Characterization of an adapted step at 11.2 mmol H2/g without any microbial population to the physicochemical or biological bioconversion of carbon monoxide into pretreatment. In comparison, the butanol using next-generation bioH2 yields on MSW were sequencing technology approximately 4-fold lower. BioH2 Guillaume Bruant, National Research production from glucose reached the Council Canada theoretical maximum for dark fermentation of 4 mol H2/mol glucose, Evaluation of second generation whereas MCC produced 9.4 mmol biofuels production from native H2/g cellulose at 7-fold higher H2 halophytes by chemical- production rates than SWG. The characterization of Salicornia sinus- consolidated bioprocessing (CPB) persica capabilities of C. saccharolyticus Mette H. Thomsen, Institute Energy present opportunities for cost savings Center of capital and operational expenses in excess of 50%. The advantages of Abstracts utilizing thermophilic microorganisms on low-cost carbon feedstocks without Brian M. Baynes prior thermo-chemical treatment for Solid catalysts, despite their broad bioH2 will be discussed. applicability as the workhorses of the petrochemical industry, have seen John Cumbers little penetration in biomass hydrolysis, because the required Tuesday, December 10, 2013 | catalytic power, lifetime, and costs 2:00pm – 3:30pm have been difficult to achieve. Here we introduce a family of novel, Feedstock Conversion chemically-defined materials that are Technologies highly potent and reusable biomass hydrolysis catalysts. Mild operating

temperature, relatively short residence time on the order of an Red Text: Invited 55

hour, and easy catalyst reuse combine processing of renewable resources will to enable biomass to be converted to be a growing economic factor in sugar and other products at very low future. The sugars after the pre- cost. Product selectivity can be treatment of several agricultural and tailored to yield (1) fermentable forestry feedstocks can be converted sugars, (2) soluble fiber, or (3) sugar by lactic acid bacteria to produce lactic dehydration products such as acid. Lactic acid, its salts and esters HMF/furfural. This talk will highlight have a wide range of potential uses Midori’s work in invention and and are extensively used in diverse development of this technology fields, e.g. bioplastics. The goal is to through 100 kg/hr pilot tests over the develop a fermentation process based past four years and its potential to on the substitution of expensive make ultra low cost sugars and other nutrients and supplements by cheaper renewable products a reality. materials from renewable resources due to their main proportion of the Joachim Venus whole process costs. Besides the basic Renewable feedstocks (e.g. crops, research projects respecting the lignocellulosics, green biomass, screening and characterization of residues) can be utilized directly, e.g. microorganisms, phenotypic as energy carriers, as packaging optimization, down-stream processing materials, as fibres, for the production of fermentation products, application of colouring agents or as lubricants. and refining of lactic acid, economic However, they can also be converted assessment of bioconversion biotechnologically by enzymes and processes the scale-up to a technical microorganisms, giving us access to a scale of several processing steps have multitude of new, biocompatible to be developed for transferable products and possible uses. Often the solutions of bioconversion 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 Red Text: Invited 56

processing of the lactic acid the separation of impurities after This presentation will discuss the use fermentation is a major process cost of next-generation sequencing too. Therefore an optimization is technology to perform microbial necessary to find a balance between community analyses of anaerobic the substitution of expensive nutrients undefined mixed cultures, with the and the limitation of interfering or objective to identify microbial species undesirable components of natural particularly adapted to the raw materials respectively. bioconversion of carbon monoxide Exploitation of high quality L(+)- and (CO), a major component of syngas, D(-) lactic acid for the production of into butanol. CO enrichment biopolymers is one of the recent experiments were performed with an applications. Conventional processes anaerobic granular sludge treating for down-streaming are based on agricultural wastes. The sludge was precipitation steps that generate large incubated during two months at amounts of chemical effluents. mesophilic temperature, with Consequently the environmental continuous CO injections in the impact of traditional processes can be headspace, creating an atmosphere of reduced by using alternative 100% CO. Liquid samples were technologies, such as electrodialysis collected after one and two months of with monopolar and bipolar operation, for subsequent microbial membranes. community analyses. Presence of volatile fatty acids (VFA) and alcohols Guillaume Bruant was verified by gas chromatography Microbial production of butanol is still after two months. DNA was extracted conventionally based on the utilization from the initial sludge and from the of carbohydrates as carbon feedstock. samples collected during enrichment 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 Red Text: Invited 57

identified in the initial sludge, as 1. Introduction: Abu Dhabi exemplifies opposed to 14 after one month and 11 a coastal desert, where seawater after two months of operation. could be used for salt-tolerant crops Bacteroidetes, Actinobacteria and (halophytes) cultivation. The produced Proteobacteria, which were the halophyte biomass could be utilized in dominant phyla in the initial sludge, feed, food and/or energy production, significantly decreased after CO depending on its chemical enrichments. On the opposite, composition. In this study the UAE Firmicutes became ultra dominant, native halophyte Salicornia sinus- representing more than 82% of the persica was studied for its potential to total bacterial sequences after two be used as a feedstock for bioethanol months. The order Clostridiales production. Fresh Salicornia sinus- became the most important in perica contains more than 65% of number, with Acetobacterium being water. For such green biomass direct the most representative genus fractionation and fermentation can be (49.8%). Archaeal diversity was less advantageous. This allows for water important since almost all the preservation and the ability to run at sequences were classified into the lower dry matter in the fermentation same phylum, Euryarchaeota, for both step. Chemical characterization and the initial anaerobic microbial ethanol potential of the juice and consortium and the CO-adapted fibers of the fractionated Salicornia population. After CO enrichments, the sinus-perica was examined in this class Methanobacteria increased, with study. Methanobacterium becoming the most representative archaeal genus (44.6% 2. Methodology: Two batches of of the total archaeal sequences). On Salicornia sinus-persica (washed and the opposite, the class unwashed) were juiced, where two 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 Mette H. Thomsen baker’s yeast. High Performance Red Text: Invited 58

Liquid Chromatography was used for achieved. This shows that fresh juice ethanol, sugars and other metabolites of Salicornia sinus-persica is a good analysis as described in (Sluiter et al., medium for yeast fermentation - but 2008b). more work is needed to identify all fermentable sugars in the juice. 3. Results: The juice fractions were found to represent 68.86 ± 1.78% of Tuesday, December 10, 2013 | the unwashed batch and 74.09 ± 4:00pm – 5:30pm 3.68% of the washed batch. The fiber fractions were found to contain Sustainability Performance 98.99% (unwashed biomass) and Value in Industrial 99.52 % DM (washed biomass) of Biotechnology Processes which 19.77% (unwashed biomass) and 19.73% (washed biomass) was Life Cycle Assessment of ash. Dry matter content of the juices Transportation Fuel Production from were found to be 13.06% (unwashed) Hydrothermal Liquefaction of Algae and 11.58% (washed) of which Grown in Open Ponds 58.01% (unwashed) and 57.37 % Moderator: Benjamin Saydah, (washed) was ash. Sugar analysis Sapphire Energy revealed relatively low concentration of glucose, xylose, and arabinose in TBD the juice fractions (8.40 g/L glu, 5.97 Matthew Rudolf g/L xyl, and 3.42 g/L ara in juice of unwashed biomass and 7.39 g/L glu Is Wood Bioenergy Carbon Neutral? 4.87 g/L xyl, and 2.17 g/L ara in juice Roger Sedjo, Resources for the Future of washed biomass) and not much difference was observed between the The Commercialization of Readily washed and unwashed biomass. The Biodegradable, Biobased Fluids and fiber fractions contained 15.6 g/100 g Cleaners, and the Motivations Driving DM glu, 11.55 g/100 g DM xyl, and Their Adoption 14.04 g/100 g DM ara for the Mike Guggenheimer, RSC Bio unwashed biomass and 16.67 g/100 g Solutions DM glu 13.01 g/100 g DM xyl, and

13.82 g/100 g DM ara for the washed Abstracts biomass. This is comparable to the lignocellulose content of the mature Benjamin Saydah (dry) plant (Cybulska et al., 2013). Life cycle assessment (LCA) has been The highest ethanol yield in SSF with used widely in recent years to Baker’s yeast was achieved in juice estimate the energy and extracted from washed biomass after environmental implications of enzymatic hydrolysis. In this deploying algae-to-energy systems at experiment 129.61% of the theoretical scale. Until recently, the emergent yield (based on sugar analysis) was nature of the industry required that Red Text: Invited 59

most LCA studies were based largely on assumptions about how the Roger Sedjo industry would develop, likely The view that wood biomass for impacting the value of the results. For energy is “carbon neutral” since its this study data from a pilot-scale use has approximately zero net facility is used to estimate the life because carbon emissions released in cycle energy and emissions profiles of its utilization for bioenergy are algae-to-biofuels production using subsequently captured in forest hydrothermal liquefaction (HTL) as the regrowth has been challenged by the conversion pathway. HTL is attractive use of a static accounting view of the because it is efficient, using the entire forest carbon. The challenge argues algae cell biomass rather than only that the use of wood for biofuels will the lipid fraction, and it is flexible, result in a decrease in the forest stock producing a nonaqueous phase liquid and a net reduction in the carbon product that can be blended during captured in the forest, at least for an petroleum refining and upgraded into extended period. The argument of this drop-in fuels, such as gasoline or paper is the static approach ignores diesel. The pilot data is used to inform dynamic economic factors. The the scale up to commercial scale and a anticipated future use of forest for LCA is conducted on the estimates of energy provides incentives to increase commercial scale operation. The pilot forestry investments in forest stocks, scale and commercial scale processes which will increase carbon are then compared to current bio-fuels sequestration in the near term and process for both their Energy Return offset carbon losses due to increased on Investment and Greenhouse Gas harvesting of forests for energy. This emissions. Results suggest that the situation is demonstrated with a algal biofuel production based on HTL dynamic optimization forest can provide substantial greenhouse management model which shows that gas (GHG) emission reductions investments in the creation of forest compared to petroleum-derived biomass can often anticipate future benchmarks. Sensitivity analyses uses and thus forest stock, and thus reveal several bottlenecks currently sequestered carbon, can actually rise holding back HTL-based algae-to- in anticipation of higher future energy production including industrial demand. Empirical support for this CO2, and nitrogen fertilizer supply outcome is provided from the forest chains, biocrude yields and heat experience of the U.S. since the mid- recycling. Efforts to address these in 1990s. industry and the implications on anticipated life cycle impacts are Mike Guggenheimer discussed. The results of readily biodegradable, biobased fluids and cleaners can be Matthew Rudolf seen across a wide range of industries TBD and segments. In solely looking at Red Text: Invited 60

performance, many readily DEINOCHEM: The Deino Way to Green biodegradable, biobased chemicals Chemicals have proven time and time again to Nagib Ward, Deinove SA be high performers in comparison to traditional petroleum-based Corbion: Developing a biobased counterparts. This single paper products portfolio submission would explore lessons Hans van der Pol, Corbion learned and successful commercialization examples in a wide Abstracts variety of industries (marine to utility fleets, waste segment and offshore oil John Perkins & gas) seeing the benefits of Royal DSM N.V. is a global science- renewable chemical solutions, such as based company active in health, readily biodegradable, biobased nutrition and materials. DSM is a functional fluids and cleaners. These frontrunner in applied industrial products are developed from biotechnology to produce ingredients ingredients from a number of regular for food and feed, or in bio-conversion BIO meeting attendees. The single technology for fuels or bio-based submission would also explore the chemicals. commercial motivations (performance, value and sustainability) driving the Today’s market needs are driven by a switch from conventional number of major global trends and petrochemicals to readily challenges. At DSM we’re using our biodegradable, biobased alternatives. innovative strengths to address some of the most important of these trends Wednesday, December 11, and challenges, such as climate 2013 | 8:30am – 10:00am change, energy independence. Advanced biofuels such as cellulosic Microbial Synthetic Pathways ethanol offer excellent solutions to to Renewable Chemicals and these challenges of today and even more for future generations. Biofuels

During the last 5-10 years a Bright Science at DSM: Development tremendous progress was made to of a Commercial Lignocellulosic continuously drive down Biomass-to-Ethanol Process manufacturing costs which posed Moderator: John Perkins, DSM major hurdles for commercialization. It has been recognized that cost Screening and Analysis Pipeline for the optimization needs an integral process Microbial Production of Renewable view (feedstock, pretreatment, Chemicals and Fuels enzymatic hydrolysis, yeast Michael Leavell, Amyris propagation and ethanol fermentation using high performing yeast) rather Red Text: Invited 61

than optimization of single DEINOVE (Alternext Paris: ALDEI) is a technologies. This presentation will cleantech company that designs, show how innovation and an develops and markets novel industrial integrated research approach are used processes using biotransforming to drive yeast and enzyme platforms based on the untapped development programs. Latest results extraordinary and advantageous to improve thermo-tolerant enzyme properties of the Deinococci. Originally mixes, their production as well as founded in France, and principally advanced yeasts with improved located in Montpellier, DEINOVE is robustness and C5 sugar converting now offering its technology platforms properties will be discussed. globally, with commercial representation in both the EU and Michael Leavell United States (www.deinove.com). Amyris is an integrated renewable products company focused on Taking advantage of the unique providing sustainable alternatives to a genetic properties and unusual broad range of petroleum-sourced robustness of this relatively unknown products. Our industrial synthetic bacterial phylum, DEINOVE optimizes biology platform converts plant sugars natural fermentation and metabolic into a variety of molecules that can be capabilities of these bacterial "micro- used in a wide range of renewable factories" to produce renewable green products, including ingredients in: chemicals. The company is structured cosmetics, flavors and fragrances, in two groups: while DEINOL is polymers, industrial lubricants, focused on producing renewable consumer products, and renewable advanced biofuels, DEINOCHEM is fuels. To enable rapid strain focused on green chemicals or development, a high-throughput strain intermediates. Of notice, two screening pipeline is needed as well as subsidiaries operate independently, an in-depth analysis of winning strains yet falling within the DEINOCHEM using omics technologies. Together group: new antibiotics research these technologies allow identification (DEINOBIOTICS SAS) and plastics and characterization of improved bioremediation (THANAPLASTTM). strains, enabling strain engineers to quickly iterate on strain designs. This DEINOCHEM is focused on producing presentation will focus on Amyris’ high renewable chemicals and reduced throughput screening platform and sugars from a variety of starting omics analysis tools, and present materials including cellulosic and other specific examples of how each renewable feedstocks. The main costs technology enables strain of biochemical production are development. concentrated on feedstock, enzymes and capital, with feedstock and Nagib Ward enzymes often accounting to 50% or more of the total cost. DEINOCHEM’s Red Text: Invited 62

platforms leverage thermophillic and metabolic properties) of organisms (45-55C) that are not DEINOCHEM’s platforms, and subject to the common constraints or DEINOCHEM's developmental research limitations often seen in industry, and results, demonstrate the huge release their own enzymes in solution potential for these microorganism’s to during the fermentation/hydrolysis, a become true bacterial micro-factories rare process advantage named for the production of bio-based Consolidated Bioprocessing (CBP). chemicals in a cost-effective, Having the ability of produce end- environmentally friendly way. products and expressing the enzymes necessary to break down feedstock for Hans van der Pol biotransformation, all at higher Corbion (Purac) is built on a strong temperatures, gives DEINOCHEM’s foundation of leading edge solutions a unique and significant cost fermentation and down-stream advantage in techno-economic processing technologies, deep market analyses. understanding, strong customer relationships and a unique global Due to the unique genetics of supply chain. Sustainability is in the Deinococci and essential towards heart of our business. Based on the biochemicals production, DEINOVE strong foundation in technology, developed specific proprietary sustainability and cooperation, molecular biology tools to both tailor Corbion is building a differentiated the end products production and biobased products portfolio. Thus, optimize the metabolism of its Corbion products are designed by platforms. DEINOVE is the World science, powered by nature, and leading synthetic biology group delivered through dedication. This leveraging Deinococci for industrial presentation will discuss the recent applications. These properties are key expansion of the Corbion products and success factors for DEINOVE’s project portfolio -into area's such as targeted industrial applications by succinic acid, FDCA and CaPropionate- allowing a platform customization for and highlight key partnerships, each commercial partner, independent capabilities and technologies that form of either the starting feedstock or the basis for the development into a pretreatment technology employed. full-fledged biobased products company. In summary, the exceptional properties (robustness, biodiversity

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