US 2008O131958A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2008/0131958 A1 Remmereit et al. (43) Pub. Date: Jun. 5, 2008

(54) ENERGY PRODUCTION WITH Publication Classification HYPERTHERMOPHILIC (51) Int. Cl. (76) Inventors: Jan Remmereit, Hovdebygda CI2M I/02 (2006.01) (NO); Michael Thomm, CI2M I/00 (2006.01) Regensburg (DE) (52) U.S. Cl...... 435/290.1; 435/252.1 Correspondence Address: MEDLEN & CARROLL, LLP 101 HOWARD STREET, SUITE 350 (57) ABSTRACT SAN FRANCISCO, CA 94105 The present invention relates to the field of degradation with (21) Appl. No.: 11/879,710 hyperthermophilic organisms, and in particular to the use of hyperthermophilic degradation to produce heat from a biom (22) Filed: Jul.18, 2007 ass. In some embodiments, a biomass is fermented in the presence of hyperthermophilic organisms to produce heat. Related U.S. Application Data The heat is used to heat a liquid which is used directly in a heat (60) Provisional application No. 60/831,635, filed on Jul. pump or radiant heat or to produce electricity or drive a steam 18, 2006. turbine. US 2008/013 1958 A1 Jun. 5, 2008

ENERGY PRODUCTION WITH the group consisting of a mineral source, Vitamins, amino HYPERTHERMOPHILIC ORGANISMS , an energy source, and a extract. 0008. In some embodiments, the energy transfer system is selected from the group consisting of a fuel cell, a 0001. This application claims the benefit of U.S. Prov. unit, a thermocouple, and a heat transfer system. In further Appl. 60/831,635 filed Jul. 18, 2006 incorporated by refer embodiments, the combustion unit comprises a steam pow ence herein in its entirety. ered system. In still further embodiments, the steam powered system is a steam turbine or generator. In some embodiments, FIELD OF THE INVENTION the heat transfer system comprises a heat pump. In some embodiments, the energy transfer system is a thermocouple 0002 The present invention relates to the field of degra and wherein the energy transfer system further comprises an dation with hyperthermophilic organisms, and in particular to electrolysis system that coverts water into hydrogen and oxy the use of hyperthermophilic degradation to produce heat gen. In some preferred embodiments, the biomass is selected from a biomass. from the group consisting of sewage, agricultural waste prod ucts like corn steep liquor and soybean hulls, brewery grain BACKGROUND OF THE INVENTION by-products, food waste, organic industry waste, forestry 0003. The cost of conventional energy sources has waste, crops, grass, seaweed, plankton, algae, fish, fish waste, increased dramatically in the last few years, and the use of and combinations thereof. many conventional energy sources such as oil, and 0009. In some embodiments, the present invention pro nuclear power has been demonstrated to be harmful to the vides methods comprising: a) providing a biomass and a environment. population of at least one genus of a hyperthermophilic 0004. Many clean alternative energy sources have been ; b) fermenting the biomass in the presence of the developed or proposed. Such sources include Solar energy, population of at least one genus of a hyperthermophilic geothermal energy, wind energy, hydroelectric energy, hydro organism under conditions such that heat is produced; c) gen reactors and fuel cells. However, many of these sources using the heat to produce electricity or heat a liquid. In some are either expensive (Solar energy) or limited by geographical embodiments, the hyperthermophilic organisms are anaero concerns (geothermal, wind and hydropower). bic hyperthermophilic organisms. In some preferred embodi 0005. Other alternative energy sources make use of biom ments, the anaerobic hyperthermophilic organisms are ass. However, those systems often involve the production of a Selected from the group consisting of the genera Pyrococcus, secondary product such as ethanol or involve combusting the Thermococcus, Palaeococcus, Acidianus (Aeropyrum and materials. These methods suffer from problems including are not anaerobic) Pyrobaculum, Pyrodictium contamination of the environment and requiring the use of Pyrolobus, Methanopyrus, Methanothermus, hyperthermo valuable farmland to produce biomass. philic Methanococci like Mc. jannaschii Fervidobacterium, and Thermotoga, and combination thereof. In other embodi 0006. Accordingly, what is needed in the art is alternative ments, the hyperthermophilic organisms are aerobic hyper systems to utilize waste biomass materials or naturally avail thermophilic organisms selected from the genera Thermus, able biomass materials to produce heat or electricity. Bacillus, and Thermoactinomyces. In still other embodi ments, the aerobic hyperthermophilic organisms are selected SUMMARY OF THE INVENTION from the group consisting of Aeropyrum permix, Sulfolobus 0007. The present invention relates to the field of degra solfataricus, sedula, Sulfobus tokodai, dation with hyperthermophilic organisms, and in particular to acidophilum and Thermoplasma volcanium, the use of hyperthermophilic degradation to produce heat and combinations thereof. In some preferred embodiments, from a biomass. In some embodiments, the present invention the biomass is selected from the group consisting of sewage, provides a system comprising: a bioreactor, the bioreactor agricultural waste products like corn steep liquor and Soybean containing biomass and a population of at least one genus of hulls, brewery grain by-products, food waste, organic indus hyperthermophilic organisms; and an energy transfer system. try waste, forestry waste, crops, grass, seaweed, plankton, In some embodiments, the hyperthermophilic organisms are algae, fish, fish waste, and combinations thereof. In some anaerobic hyperthermophilic organisms. In some preferred embodiments, the biomass is Supplemented with a cell culture embodiments, the anaerobic hyperthermophilic organisms media component selected from the group consisting of a are selected from the group consisting of the genera Pyrococ mineral Source, vitamins, amino acids, an energy source, and cus, Thermococcus, Palaeococcus, Acidianus, Pyrobaculum, a microorganism extract. Pyrodictium, Pyrolobus, Methanopyrus, Methanothermus, 0010. In some embodiments, the liquid is water and the hyperthermophilic Methanococci like Mc. jannaschii, Fervi heating produces steam. In some embodiments, the Steam is dobacterium and Thermotoga, and combination thereof. In used to drive a steam turbine to produce electricity. In further other embodiments, the hyperthermophilic organisms are embodiments, the heated liquid is transferred to a building for aerobic hyperthermophilic organisms selected from the gen radiant heat. In some embodiments, the electricity is pro era Thermus, Bacillus, and Thermoactinomyces. In still other duced via a thermocouple. In further embodiments, the elec embodiments, the aerobic hyperthermophilic organisms are tricity is used for electrolysis of water. In some embodiments, selected from the group consisting of Aeropyrum permix, the liquid is transferred to a heat pump. Metallosphaera sedula and other Metallosphaera species 0011. In some embodiments, the present invention further Sulfolobus solfataricus, Sulfobus tokodai, Thermoplasma provides methods comprising: a) providing a biomass and a acidophilum and Thermoplasma volcanium, and combina population of at least one genus of a hyperthermophilic tions thereof. In some embodiments, the biomass is Supple organism; and b) degrading the biomass in the presence of the mented with a cell culture media component selected from population of at least one genus of a hyperthermophilic US 2008/013 1958 A1 Jun. 5, 2008 organism under conditions such that degradation products are tural crop residues, energy plantations, and municipal and produced. In some preferred embodiments, the anaerobic industrial wastes. The term “biomass, as used herein, hyperthermophilic organisms are selected from the group excludes components of traditional media used to culture consisting of the genera Pyrococcus, Thermococcus, Palaeo , such as purified starch, peptone, yeast coccus, Acidianus, Pyrobaculum, Pyrolobus, Pyrodictium, extract but includes waste material obtained during industrial Methanopyrus, Methanothermus, hyperthermophilic Metha processes developed to produce purified Starch. According to nococci like Mc. jannaschii Fervidobacterium and Thermo the invention, biomass may be derived from a single source, toga, and combination thereof. In other embodiments, the or biomass can comprise a mixture derived from more than hyperthermophilic organisms are aerobic hyperthermophilic one source; for example, biomass could comprise a mixture organisms selected from the genera Thermus, Bacillus, and of corn cobs and corn Stover, or a mixture of grass and leaves. Thermoactinomyces. In still other embodiments, the aerobic Biomass includes, but is not limited to, bioenergy crops, hyperthermophilic organisms are selected from the group agricultural residues, municipal Solid waste, industrial Solid consisting of Aeropyrum permix, Sulfolobus solfataricus, Sul waste, sludge from paper manufacture, yard waste, wood and fobus tokodai, Metallosphaera sedula, Thermoplasma aci forestry waste. Examples of biomass include, but are not dophilum and Thermoplasma volcanium, and combinations limited to, corn grain, corn cobs, crop residues such as corn thereof. In some preferred embodiments, the biomass is husks, corn Stover, corn steep liquor, grasses, wheat, wheat selected from the group consisting of sewage, agricultural Straw, barley, barley Straw, grain residue from barley degra waste products, brewery grain by-products, food waste, dation during brewing of beer, hay, rice Straw, Switchgrass, organic industry waste, forestry waste, crops, grass, Seaweed, waste paper, cane bagasse, Sorghum, Soy, components plankton, algae, fish, fish waste, and combinations thereof. In obtained from processing of grains, trees, branches, roots, Some embodiments, the biomass is Supplemented with a cell leaves, wood chips, sawdust, shrubs and bushes, soybean culture media component selected from the group consisting hulls, vegetables, fruits, flowers and animal manure. In one of a mineral Source, Vitamins, amino acids, an energy source, embodiment, biomass that is useful for the invention includes and a microorganism extract. In some further preferred biomass that has a relatively high carbohydrate value, is rela embodiments, the degradation products are selected from the tively dense, and/or is relatively easy to collect, transport, group consisting of hydrogen, methane and ethanol. In some store and/or handle. embodiments, the methods further comprise the step of con 0014. As used herein, the term “biomass by-products' Verting the degradation products into energy. In some refers to biomass materials that are produced from the pro embodiments, the methods further comprise the step of using cessing of biomass. the hydrogen in a fuel cell. In some embodiments, the meth 0015. As used herein, the term “bioreactor” refers to an ods further comprise the step of using the methane or ethanol enclosed or isolated system for containment of a microorgan in a combustion unit. ism and a biomass material. The “bioreactor” may preferably 0012. In some embodiments, the present invention pro be configured for anaerobic growth of the microorganism. vides methods for reducing dioxide emissions com 0016. As used herein, the term “hyperthermophilic organ prising: a) providing a biomass and a population of at least ism” means an organism which grows optimally attempera one genus of a hyperthermophilic organism; b) anaerobically tures above 80° C. degrading said biomass in the presence of said population of 0017. As used herein, the terms “degrade' and “degrada at least one genus of a hyperthermophilic organism to pro tion” refer to the process of reducing the complexity of a duce Substrates for energy production; and c) producing Substrate. Such as a biomass Substrate, by a biochemical pro energy from said Substrates, wherein emis cess, preferably facilitated by microorganisms (i.e., biologi sions are reduced as compared to aerobic degradation of said cal degradation). Degradation results in the formation of sim biomass materials. In further embodiments, the present pler compounds such as methane, ethanol, hydrogen, and invention provides methods for generating carbon credits other relatively simple organic compounds (i.e., degradation comprising: a) providing a biomass and a population of at products) from complex compounds. The term "degradation' least one genus of a hyperthermophilic organism; b) anaero encompasses anaerobic and aerobic processes, including fer bically degrading said biomass in the presence of said popu mentation processes. lation of at least one genus of a hyperthermophilic organism to produce Substrates for energy production, and c) producing DETAILED DESCRIPTION OF THE INVENTION energy from said Substrates under conditions such that carbon credits are generated. 0018. The present invention relates to the field of biomass degradation with hyperthermophilic organisms, and in par DEFINITIONS ticular to the use of hyperthermophilic degradation to pro duce heat from a biomass. For convenience, the description of 0013. As used herein, the term “biomass” refers to bio the invention is provided in the following section: A. Hyper logical material which can be used as fuel or for industrial thermophilic organism; B. Biomass; C. Degradation and production. Most commonly, biomass refers to plant matter energy production; and D. Carbon credit generation. grown for use as biofuel, but it also includes plant or animal matter used for production of fibers, chemicals or heat. Bio A. Hyperthermophilic Organisms mass may also include biodegradable wastes that can be used as fuel. It is usually measured by dry weight. The term bio 0019. The present invention comtemplates the use of mass is useful for plants, where some internal structures may hyperthermophilic organism for fermenting biomass. Ther not always be considered living tissue, such as the wood mophilic bacteria are organisms which are capable of growth (secondary xylem) of a tree. This biomass became produced at elevated temperatures. Unlike the mesophiles, which grow from plants that convert Sunlight into plant material through best at temperatures in the range of 25-40°C., or psychro photosynthesis. Sources of biomass energy lead to agricul philes, which grow best attemperatures in the range of 15-20° US 2008/013 1958 A1 Jun. 5, 2008

C., thermophiles grow best attemperatures greater than 50° Stearothermophilus, Anaerocellum thermophilus, Thermoac C. Indeed, some thermophiles grow best at 65-75° C., and tinomycees vulgaris, and members of the order Thermoto Some of the grow at temperatures up to gales, including, but not limited to Thermotoga elfei, Ther 113°C. (See e.g., J. G. Black, Microbiology Principles and motoga hypogea, Thermotoga maritima, Thermotoga Applications, 2d edition, Prentice Hall, New Jersey, 1993 p. neapolitana, Thermotoga subterranean, Thermotoga ther 145-146; Dworkin, M., Falkow, S. Rosenberg, E. Schleifer, marum, Petrotoga miotherma, Petrotoga mobilis, Thermosi K-H. Stackebarndt E. (eds) The prokaryotes, third edition, pho africanus, Thermosipho melanesiensis, Fervidobacte volume 3, p. 3-28296 and p. 797-814 and p. 899-924; Madi rium islandicum, Fervidobacterium nodosum, gan M., Martinko, J. Brock Biology of Microorganisms, elev Fervidobacterium pennavorans, Fervidobacterium gond enth edition, p. 430-441 and 414-415). wanense, Geotoga petraea, Geotoga subterranea. 0020. The thermophilic bacteria encompass a wide variety 0022. In some embodiments, hyperthermophilic strains of of genera and species. There are thermophilic representatives the above organisms suitable for fermenting biomass will be included within the phototrophic bacteria (i.e., the purple selected by Screening and selecting for Suitable strains. In still bacteria, green bacteria, and cyanobacteria), bacteria (i.e., further embodiments, suitable strains will be genetically Bacillus, Clostridium, Thiobacillus, Desulfotomaculum, modified to include desirable metabolic enzymes, including, Thermus, Lactic bacteria, Actinomycetes, Spirochetes, but not limited to hydrolytic enzymes, proteases, alcohol and numerous other genera), and many hyperthermophilic dehydrogenase, and pyruvate decarboxylase. See, e.g., (Bra/ orders (i.e., Pyrococcus, Thermococcus, Thermotoga, Sul u, B., and H. Sahm 1986 Arch. Microbiol. 146:105-110: folobus, and some methanogens). There are aerobic as well as Bra?u, B. and H. Sahm 1986 Arch. Microbiol. 144:296-301; anaerobic thermophilic organisms. Thus, the environments in Conway, T.Y.A. Osman, J. I. Konnan, E. M. Hoffmann, and which thermophiles may be isolated vary greatly, although all L. O. Ingram 1987. J. Bacteriol. 169:949-954; Conway, T., of these organisms are isolated from areas associated with G. W. Sewell, Y. A. Osman, and L. O. Ingram 1987 J. high temperatures. Natural geothermal habitats have a world Bacteriol. 169:2591-2597; Neale, A. D., R. K. Scopes, R. E. wide distribution and are primarily associated with tectoni H. Wettenhall, and N. J. Hoogenraad (1987 Nucleic Acid. cally active Zones where major movements of the earth's crust Res. 15:1753-1761; Ingram, L. O., and T. Conway 1988 occur. Thermophilic bacteria have been isolated from all of Appl. Environ. Microbiol. 54:397-404; Ingram, L. O. T. the various geothermal habitats, including boiling springs Conway, D. P. Clark, G. W. Sewell, and J. F. Preston 1987 with neutral pH ranges, -rich acidic springs, and deep Appl. Environ. Microbiol. 53:2420-2425). In some embodi Sea Vents. In general, the organisms are optimally adapted to ments, a PET operon is introduced into the . the temperatures at which they are living in these geothermal See U.S. Pat. No. 5,000,000, incorporated herein by reference habitats (T. D. Brock, “Introduction: An overview of the in its entirety. thermophiles, in T. D. Brock (ed.), Thermophiles. General, 0023. In some embodiments, hyperthermophiles that pro Molecular and Applied Microbiology, John Wiley & Sons, duce ethanol via degradation are selected. In some embodi New York 1986, pp. 1-16: Madigan M., Martinko, J. Brock ments, such hyperthermophiles are selected in media contain Biology of Microorganisms, eleventh edition, p. 442-446 and ing progressively higher amounts of ethanol to select for p. 299-328). Basic, as well as applied research on thermo strains with increased ethanol tolerance. Accordingly, some philes has provided some insight into the physiology of these embodiments of the present invention provide hyperthermo organisms, as well as promise for use of these organisms in philes with increased ethanol tolerance or increased ability to industry and biotechnology. produce ethanol. In some preferred embodiments, the hyper 0021. The present invention is not limited to the use any thermophiles utilize lignocellulosic biomass. In further pre particular hyperthermophilic organism. In some embodi ferred embodiments, the hyperthermophile utilize glucose, ments, mixtures of hyperthermophilic organisms are utilized. Xylose, arabinose, galactose, and mannose. In some embodiments, the hyperthermophiles are from the archaeal order Thermococcales, including but not limited to B. Biomass hyperthermophiles of the genera Pyrococcus, Thermococcus, and Palaeococcus. Examples of particular organisms within 0024. The present invention contemplates the degradation these genera include, but are not limited to, Pyrococcus furio of biomass with hyperthermophilic organisms. The present sus, Thermococcus barophilus, T. aggregans, Taegaeicus, T. invention is not limited to the use of any particular biomass. litoralis, Talcaliphilus, Tsibiricus, T. atlanticus, T. siculi, T. Suitable biomass includes, but is not limited to, sewage, agri pacificus, T. Waiotapuensis, T Zilligi, T. guaymasensis, T. cultural waste products, brewery grain by-products, food filmicolans, T gorgonarius, T celer: T. barossii, T. hydrother waste, organic industry waste, forestry waste, crops, grass, malis, T. acidaminovorans, T. prfiindus, T. Stetteri, T. seaweed, plankton, algae, fish, fish waste, and combinations kodakaraenis, T. peptonophilis. In some embodiments, aero thereof. In some embodiments, the biomass is harvested par bic hyperthermophilic organisms such as Aeropyrum permix, ticularly for use in hyperthermophilic degradation processes, Sulfolobus solfataricus, Metallosphaera sedula, Sulfobus while in other embodiments waste or by-products materials tokodai, and Thermoplasma from a pre-existing industry are utilized. volcanium are utilized. While in other embodiments, anero 0025. In some preferred embodiments, the biomass is bic or facultative aerobic organisms such as Pyrobaculum lignocellulosic. In some embodiments, the biomass is pre calidifontis and Pyrobaculum Oguniense are utilized. Other treated with cellulases or other enzymes to digest the cellu useful archaeal organisms include, but are not limited to, lose. In some embodiments, the biomass is pretreated by Sulfolobus acidocaldarius and Acidianus ambivalens. In heating in the presence of a mineral acid or base catalyst to Some embodiments, the hyperthermophilic organisms are completely or partially hydrolyze hemicellulose, decrystal bacteria, Such as Thermus aquaticus, Thermus thermophilus, lize cellulose, and remove lignin. This allows cellulose Thermus flavu, Thermus ruber, Bacillus caldotenax, Bacillus enzymes to access the cellulose. US 2008/013 1958 A1 Jun. 5, 2008

0026. In still other preferred embodiments, the biomass is ments, proteins, acids and glycerol are formed which can be Supplemented with minerals, energy sources or other organic purified for other uses or, for, example, used as animal feeds. Substances. Examples of minerals include, but are not limited, 0030. In some embodiments, the degradation products are to those found in seawater such as NaCl, MgSO4x7H2O, removed from the vessel. It is contemplated that the high MgClx6H2O, CaClx2H2O, KCl, NaBr, HBO, and SrClx temperatures at which the degradation can be conducted 6H) and other minerals such as MnSO4xH2O, FeSOax facilitate removal of valuable degradation products from the 7H2O, CoSOx7H2O, ZnSOx7H2O, CuSOx5H2O, KAl vessel in the gas phase. In some embodiments, methane, (SO)2x12H2O, Na MoOSOX2H2O, (NHSO)2Ni(SO)x hydrogen and/or ethanol are removed from the vessel. In 6H2O, NaWOx2H2O and NaSeO. Examples of energy Some embodiments, these materials are moved from the ves Sources and other Substrates include, but are not limited to, sel via a system of pipes so that the product can be used to purified Sucrose, fructose, glucose, starch, peptone, yeast generate power or electricity. For example, in Some embodi ments, methane or ethanol are used in a combustion unit to extract, amino acids, nucleotides, nucleosides, and other generate power or electricity. In some embodiments, steam components commonly included in cell culture media. power is generated via a steam turbine or generator. In some embodiments, the products are packages for use. For C. Degradation and Energy Production example, the ethanol, methane or hydrogen can be packaged 0027. In preferred embodiments of the present invention, in tanks or tankers and transported to a site remote from the one or more populations of hyperthermophilic organisms are fermenting vessel. In other embodiments, the products are fed utilized to degrade biomass. In some embodiments, the bio into a pipeline system. mass is transferred to a vessel Such as a bioreactor and inocu 0031. In still other embodiments, heat generated in the lated with one or more strains of hyperthermophilic organ vessel is utilized. In some embodiments, the heat generated is isms. In some embodiments, the environment of the vessel is utilized in radiant system where a liquid is heated and then maintained at a temperature, pressure, and pH Sufficient to circulated via pipes or tubes in an area requiring heating. In allow the strain(s) to metabolize the feedstock. In some pre Some embodiments, the heat is utilized in aheat pump system. ferred embodiments, the environment has no added sulfur or In still other embodiments, the heat is utilized to produce inorganic Salts or is treated to remove or neutralize electricity via a thermocouple. In some embodiments, the Such compounds. In some preferred embodiments, the envi electricity produced is used to generate hydrogen via an elec ronment is maintained at a temperature above 45° C. In still trolysis reaction. further embodiments, the environment the environment is maintained at between 55° C. and 90° C. In some preferred D. Carbon Credit Trading embodiments, , starches, Xylans, celluloses, oils, petro 0032. In some embodiments, the present invention pro leums, bitumens, amino acids, long-chain fatty acids, pro vides methods for generating carbon credits for trading in teins, or combinations thereof, are added to the biomass. In established carbon credit trading programs such as those Some embodiments, water is added to the biomass to forman established under the Kyoto protocol. The European Union at least a partially aqueous medium. In some embodiments, Emission Trading System (EU ETS), which began operation the aqueous medium has a dissolved oxygen gas concentra in January 2005, is the largest multi-national, multi-sector tion of between about 0.2 mg/liter and 2.8 mg/liter. In some greenhouse gas emissions trading scheme in the world. The embodiments, the environment is maintained at a pH of system was set up as the EU's response to the Kyoto Protocol between approximately 4 and 10. In some embodiments, the to the United Nations Framework Convention on Climate environment is preconditioned with an inert gas selected from Change which was negotiated in 1997 and ratified in 2005. It a group consisting of , carbon dioxide, helium, neon, is a commitment among participating industrialised nations argon, krypton, Xenon, and combinations thereof. While in to curb the rise in global temperature by abating their emis other embodiments, oxygen is added to the environment to sions of six greenhouse gases including carbon dioxide, Support aerobic degradation. methane, nitrous oxide, Sulfur hexafluoride, perfluorocarbons 0028. In some embodiments, where lignocellulosic mate (PFCs) and hydrofluorocarbons (HFCs). To date, 162 nations rial are utilized, the cellulose is pre-treated as described have ratified the agreement. Notable exceptions are the above. The pre-treated cellulose is enzymatically hydrolyzed United States and Australia. Furthermore, two of the fastest either prior to degradation in sequential saccharification and growing economies, India and China, are not required to degradation or by adding the cellulose and hyperthermophile reduce their carbon emissions under the current agreement. inoculum together for simultaneous saccharification and deg 0033. The Kyoto Protocol provides three implementation radation. mechanisms to regulate greenhouse gas emissions. The first, 0029. It is contemplated that degradation of the biomass International Emissions Trading (IET), permits countries will both directly produce energy in the form of heat as well below their current emissions limits to sell their excess allow as produce products that can be used in Subsequent processes, ances to other countries on the open market. The second, Joint including the production of energy. In some embodiments, Implementation (JI), allows investors from industrialised hydrogen, methane, and ethanol are produced by the degra countries financing greenhouse gas emissions reduction dation and utilized for energy production. In preferred projects in other industrialised countries to receive emission embodiments, these products are removed from the vessel. It credits called "emissions reduction units” (ERUs). The third, is contemplated that removal of these materials in the gas Clean Development Mechanism (CDM), lets investors from phase will be facilitated by the high temperature in the culture industrialised countries accumulate "certified emission vessel. These products may be converted into energy by stan reduction units” (CERs) for helping finance carbon reduction dard processes including combustion and/or formation of projects in developing countries. steam to drive steam turbines or generators. In some embodi 0034. The EU ETS exists in two phases and encompasses ments, the hydrogen is utilized in fuel cells. In some embodi all of the high use energy and power sectors. The first phase, US 2008/013 1958 A1 Jun. 5, 2008

which started in 2005 and will end in 2007, allows for the released from biomass by about six-fold as compared to aero trade of CO allowances with the potential to expand into the bic degradation or fermentation processes. other five greenhouse gasses. So far, it has set caps on the 0041. In some embodiments, the present invention pro emissions of 12,000 to 15,000 industrial installations across vides a system wherein energy is produced by degradation of Europe. It covers 45% of emission activities including power, biomass with hyperthermophilic organisms, and resulting concrete, pulp, paper, and ferrous metals. The second phase, carbon credits generated through the use of the system are from 2008 to 2012, could possibly coverallgreenhouse gases used to offset greenhouse gas emissions by conventional and installations, and will include JI and CDM credits in the energy production systems such as combustion of coal, natu market. It is important to note that in the first phase an amend ral gas, and oil. In some embodiments, the energy production ment called the Linking Directive was implemented which systems are under the control of a single entity, while in other enabled installations to use CERs and ERUs from JI and embodiments, the energy production systems are under the CDM to meet their emission targets. control of separate entities and the carbon credits are pur 0035. The EU ETS is monitored and regulated by the EU chased by or traded to the entity generating power by conven Commission (EUC). In both phases, the EUC places limita tional means with fossil fuels. tions on GHG which are satisfied through the trading of EU emission allowances (EUAS). The goal is to force companies EXPERIMENTAL to find the lowest cost of abatement by decreasing their GHG internally and selling any unused EUAS into the market. Dur 1. Selection of Hyperthermophilic Organisms for Degrada ing the first phase, the EUC imposes a penalty of 640 perton tion Processes of CO2 for installations that emit more than their target limit. 0042. In this example, strains of hyperthermophilic organ In addition, these installations must acquire their excess emis isms from the genera Pyrococcus, Thermococcus, Palaeococ sions in the market. This penalty will go to 6100 per ton of cus, Aeropyrum permix, Sulfolobus, Pyrobaculum, Pyrolobus, CO2 in the second phase. Pyrodictium, Thermus, Bacillus Stearothermophilus, Metal 0036 Participating countries in the EU ETS submit their losphaera, Anaerocellum, Thermoactinomyces, Thermotoga, target GHG reductions through National Allocation Plans Fervidobacterium and Geotoga are selected and screened for (NAPs) which then are approved by the EUC. According to the ability to produce fermentation byproducts ethanol, the Norwegian consultant Point Carbon, during the first phase methanol and hydrogen. Briefly, seed inoculums are prepared of the EU ETS, the EUC approved circa 6.3 billion allow by culturing the cells in YT medium (yeast extract 2.0 g/li ances and allowed for another 2.1 billion to be distributed ter, tryptone 4.0 g/liter, NaSO, 0.61 g/liter, and ASN each year. III salts) for 48 h. Flasks containing base medium (tryptone 0037. As one example of an established system, the Euro (4.0 g/liter), NaSO (0.61 g/liter), and ASN-III salts (artifi pean Bank for Reconstruction and Development (EBRD) and cial seawater salts containing NaCl 29.8 g/liter, MgCl, 1.1 the European Investment Bank (EIB) established the Multi g/liter, MgSO 2.0 g/liter, CaCl2 0.45 g/liter, KCl 0.6 lateral Carbon Credit Fund (MCCF) for countries from Cen g/liter, and NaCO 0.024 g/liter)(pH 7.0)) supplemented tral Europe to Central Asia. with specific carbohydrates (glucose, Xylose, arabinose, 0038. By joining the MCCF, private and public companies galactose, and/or mannose) (3.0 g/liter) are inoculated with as well as EBRD and EIB shareholder countries can purchase 10% seed inoculums. The flasks are then purged with prepu carbon credits from emission reduction projects financed by rified N and the incubation is carried out at 90° C.-110°C. in the EIB or EBRD to meet their mandatory or voluntary green a rotary shaker at 150 rpm. Cell growth is observed by moni house gas (GHG) emission reduction targets. toring optical density at 660 nm (ODeo). Samples are col 0039. In addition to the project credits, countries can also lected from the headspace and culture medium and analyzed participate via the MCCF in green investment schemes. This by GC for fermentation products. is an innovative way to facilitate government-to-government 2. Growth of Pyrococcus firiosus and Thermotoga maritima trade in carbon credits, whereby the selling country uses the on Waste Materials and Biomass Substrates revenue from the sale of carbon credits to support investments 0043. The hyperthermophilic archaeon Pyrococcus firio in climate-friendly projects. Carbon credits can be generated sus (growth range 67-103°C., optimal growth at 100°C.) uses from a large variety of project types, all of which reduce or simple and complex carbohydrates and converts them to avoid GHG emissions. These include credits produced from acetate, to C0, and to H. Only in the presence of elemental renewable energy Such as wind, hydro, biogas (from landfills/ sulphur (S), H is used to reduce sulphur to H.S. An expo wastewater) and biomass. nentially growing culture produces ~1 umol ml'h'. He 0040. In some embodiments, the present invention gener (Schut et al., 2007, J. Bacteriol 189, 4431-4441). Growth ates carbon credits for trading by utilizing biomass. In other experiments in the laboratory have shown that the strain embodiments, the present invention generates carbon credits requires peptone and yeast extract (as protein and Vitamin for trading by utilizing materials that would otherwise create source) in addition for good growth (2.2x10 cells/ml). On methane that is Subsequently released into the atmosphere, starch as sole carbon source only poor growth was observed Such as manure, sewage, waste water, landfilled materials and (-5x10 cells/ml). the like. The present invention is not limited to any particular 0044) Thermotoga maritima is an obligately anaerobic mechanism of action. Indeed, an understanding of the mecha hyperthermophilic bacterium growing between 55-90° C. nism of action is not needed to practice the present invention. (growth optimum at 80°C.). Like Pyrococcus it is of marine Nevertheless, it is contemplates that the use of hyperthermo origin and is cultivated in media resembling seawater. Ther philic organisms in an anaerobic degradation process is motoga is an obligate preferentially fermenting highly efficient for reducing carbon emissions, and in particu carbohydrates or complex organic matter. Fermentation of lar emissions of carbon dioxide. In particular, the use of glucose by cell Suspensions of Thermotoga yielded 118 mol anaerobic degradation reduces the amount carbon dioxide L-(+) lactate, 47 molacetate, 54 mol C0 and 9 mol H. (Huber US 2008/013 1958 A1 Jun. 5, 2008

et al., 1986, Arch. Microbiol. 144, 324-333). Some of the detect significant ethanol formation. For ethanol production, members of the Thermotogales like Fervidobacterium Fervidobacterium strains (F. nodosum and F islandicum) nodosum (Patel et al., 1985 Arch. Microbiol. 141, 63-69) and may be utilized. Fervidobacterium islandicum (Huber et al., Arch. Microbiol. 1990, 154, 105-111 have been described to produce also ethanol. F. nodosum forms after 13 h growth on glucose -25 Heat Production During Growth umol ethanol per 10 ml culture broth (Patel et al., 1985). A 0048. The measurement of energy release using a standard quantitative analysis of fermentation products (micromole of fermentor was difficult. When Pyrococcus was growing in the product formed per 10 ml culture) of T. nodosum grown on fermentor an input of 1060 Wh was required during an incu glucose revealed: Ethanol 10, acetate 115, lactate 162, CO 120 and H. 160 per 133 micromol glucose consumed. bation time of 30 h to keep the temperature of the 101 fer 0045 Both organisms do not completely oxidize organic mentor constantly at 90° C. In the absence of growing Pyro matter to CO. The carbon of the substrate is in part converted coccus cells the energy input in 30 h was 1140 Wh. This to Soluble compounds like acetae and lactate. Both organism indicates an energy input of 35.5 W per hour in the absence of produce low amounts of hydrogen and soluble compounds growing cells and 32.5 W per hour in the presence of growing like acetate. Some members of the Thermotogales have been cells. When the heat production was measured during growth described to produce ethanol in addition (Fervidobacterium). of Thermotoga no energy release by growing cells could be Thus these anaerobic organisms have the potential to synthe detected, although the microorganisms grew quite well up size energy rich compounds like H2 and ethanol. The amount within 13.5 hours to a cell density of 4x10 cells/ml. of CO produced during anaerobic degradation of biomass is significantly lower than CO release during aerobic processes 0049. It is known that large fermentors used for biotech which lead to complete oxidation of organic matter to CO. nological processes like ethanol fermentation by yeast Methane formation will not occur during this process when require cooling due to the energy released by growing yeast. pure cultures are used or when the waste substrate is steril To control the system for the detection of heat production we ized. Otherwise methane might beformed from the end prod grew yeast anaerobically at 30° C. During 95 h after inocu ucts formed by degradation of organic matter from Thermo lation of the medium no external energy input was required to toga and Pyrococcus (H/CO and acetate). Acetate can be keep the growth temperature at 30°C. and the temperature of also converted to methane but no hyperthermophilic metha the culture medium was even increased by 0.5°C. This find nogen growing on acetate has been described. Therefore, it is ing Suggests that the detecting system is suitable to measure unlikely that methane is formed from acetate when the fer energy release by microorganisms. To confirm the validity of mentation will be conducted attemperatures between 80 and our measurement it is advisable to repeat the experiment in an 100° C. air conditioned room (room temperature fixed at 20° C.). 0046. The objective was to investigate the potential of P firiosus and T. maritima as model systems for the degrada 3. Pyrococcus Furiosus /2 SME Medium tion of waste products and to investigate their ability to pro duce and to release heat during growth. The degradation of various waste products was studied in 100 1 batch cultures. A SME The energy release during growth was measured in a 101 glass fermentor. The heating system of this fermentor was modified Component Amount to lower the input of energy. The fermentor was isolated by SME 500.0 ml the use of an aluminium containing shell and further isolated KH2PO 0.5 g. by styrene. As a control, heat release by a 10 1 culture of Wolfe's mineral elixir 1.0 ml Saccharomyces cerevisiae was also measured using this sys 10xpH 6.5/new + T tem. ResaZurin, 0.1% solution 1.0 ml H2O 2 x distilled, add to a final volume 1000.0 ml of

Utilization of waste substrates Pyrococcus Thermotoga Grain residues no growth poor growth (from a brewery) (8 x 106ml) Synthetic Seawater - SME Mixture of grain residues no growth good growth Component Amount concentration and whey 1.4 x 10' no pH control 3.2 x 108w? pH control NaCl 27.7 g 473.99 mM Mixture of grain residues 1 x 10 2 x 108 MgSO4 x 7H2O 7.0 g 28.4 mM and fish innards MgCl2 x 6H2O 5.5 g. 27.1 mM Mixture of soluble starch ~1 x 10 not analyzed CaCl2 x 2H2O 0.75 g 5.1 mM and whey (final cell density KCI 0.65 g 8.7 mM was not determined) NaBr 0.1 g O.97 mM HBO 0.03 g O.49 mM SrCl2 x 6H2O 0.015 g O.056 mM Detailed formulations of the culture media are provided KJ-Lsg., 0.05% ig 0.1 ml 0.30 M below. H2O 2 x distilled, add to a final volume 1000.0 ml 0047. Since ethanol production has been described for of Some members of the Thermotogales we assayed also ethanol formation during growth on several substrates. We could not US 2008/013 1958 A1 Jun. 5, 2008

-continued Wolfe's mineral elixir 10x/pH 6.5/new + Titriplex Synthetic Seawater - SME Component amount concentration Compound Amount Concentration Titriplex 1 (Nitrillotriacetic acid) 15.0 g 78.50 nM HBO 0.03 g O.49 mM MgSO4 x 7H2O 30.0 g 121.70 mM SrCl2 x 6H2O 0.015 g O.056 mM MnSOx H2O 5.0 g 29.60 mM KJ-solution., 0.05% (w/v) 0.1 ml 0.30 M NaCl 10.0 g 171.10 mM HO 2 x distilled, add to a final volume 1000.0 ml FeSO x 7H2O 1.0 g 3.60 mM of CoSO x 7H2O 1.8 g. 6.40 mM CaCl2 x 2H2O 1.0 g 6.80 mM ZnSO x 7H2O 1.8 g. 6.30 mM CuSO x 5H2O 0.1 g O.40 mM KAI (SO)2 x 12H2O 0.18 g O.38 mM Wolfe's mineral elixir 10x/pH 6.5/new + Titriplex HBO 0.1 g 1.62 mM Na2MoC) x 2H2O 0.1 g O.41mM Compound amount concentration (NH4)2Ni(SO)2 x 6H2O 2.80 g 7.09 mM NaWO x 2H2O 0.1 g O.30 mM Titriplex 1 (Nitrillotriacetic acid) 15.0 g 78.50 nM Na-SeO. 0.1 g O.53 mM MgSO x 7H2O 30.0 g 121.70 mM H2O add to a final volume of 1000.0 ml MnSOX HO 5.0 g 29.60 mM NaCl 10.0 g 171.10 mM FeSO x 7H2O 1.0 g 3.0 mM In standard medium, the following organic Substrates were CoSO x 7H2O 1.8 g. 6.40 mM added: CaCl2 x 2H2O 1.0 g 6.80 mM ZnSO x 7H2O 1.8 g. 6.30 mM CuSO x 5HO 0.1 g O.40 mM KAl(SO4)2 x 12H2O 0.18 g O.38 mM Component Amount HBO 0.1 g 1.62 mM Na2MoC) x 2H2O 0.1 g O.41mM Yeast extract (Difco) O.1% (NH)-Ni(SO) x 6H2O 2.80 g 7.09 mM Pepton from casein (Difco) O.1% NaWOx 2H2O 0.1 g O.30 mM Starch (Merck) O.1% Na2SeO. 0.1 g O.53 mM H2O 2 x distilled, add to a final volume 1000.0 ml 0050 For Pyrococcus furiosus: pH: 7.0 of 0051 Headspace: N/CO, 0052 To study utilization of waste products we replaced For growth of Thermotoga maritima the following organic the organic components of the medium by various waste substrates were added: materials: grain residues: 5%; whey 10%; fish innards 0.95% 4. Thermotoga MM-I-Medium Compound amount Starch (Merck 1.01252.1000) O.05% 0053 Yeast extract (Difco) O.05% To study growth on waste products the organic Substrates MM-I-medium were replaced by: grain residues (5% w/w), whey 10% (v/v) Compound Amount and homogenized fish innards 0.9% (950 g/100 l). 0054 pH: 7.0 SME 250.0 ml 0055 headspace: N. KH2PO 0.5 g. (NH4)2SO 0.5 g. 0056. In some experiments first growth of Pyrococcus was NaHCO, 0.1 g studied at 90°C., if Pyrococcus failed to grow or after growth Wolfe's mineral elixir, 1.5 ml of Pyrococcus to 1x10 cells/ml the medium was cooled 10xpH 6.5/new + T down to 80° C. and then the same medium was inoculated ResaZurin, 0.1% solution 1.0 ml with Thermotoga. On the Substrate mixture grain residues and NaS x 7-9HO 0.5 g. fish innards good growth of Thermotoga was observed under H2O 2 x distilled, add to a final volume 1000.0 ml these conditions; this indicates that Thermotoga grows well in of Pyrococcus medium. 1. A system comprising: a bioreactor, said bioreactor containing biomass and a population of at least one genus of hyperthermophilic organisms: Synthetic Seawater - SME an energy transfer system. 2. The system of claim 1, wherein said hyperthermophilic Compound Amount Concentration organisms are anaerobic hyperthermophilic organisms. NaCl 27.7 g 473.99 mM 3. The system of claim 2, wherein said anaerobic hyper MgSO4 x 7H2O 7.0 g 28.4 nM thermophilic organisms are selected from the group consist MgCl2 x 6H2O 5.5 g. 27.1 mM ing of the genera Pyrococcus, Thermococcus, Acidianus, CaCl2 x 2H2O 0.75 g 5.1 mM Palaeococcus, Thermoplasma, Pyrobaculum, Pyrolobus, KC 0.65 g 8.7 mM Pyrodictium, Methanothermus, Methanopyrus, hyperther NaBr 0.1 g O.97 mM mophilic Methanococci like Mc. jannaschii, Fervidobacte rium and Thermotoga, and combination thereof. US 2008/013 1958 A1 Jun. 5, 2008

4. The system of claim 1, wherein said hyperthermophilic 22. The method of claim 16, wherein said liquid is water organisms are aerobic hyperthermophilic organisms. and said heating produces steam. 5. The system of claim 4, wherein said aerobic hyperther 23. The method of claim 22, wherein said steam is used to mophilic organisms are selected from the group consisting of drive a steam turbine to produce electricity. Aeropyrum permix, Sulfolobus solfataricus, Sulfobus toko 24. The method of claim 16, wherein said heated liquid is daii, Metallosphaera sedulla, Thermoplasma acidophilum transferred to a building for radiant heat. 25. The method of claim 16, wherein said electricity is and Thermoplasma volcanium, and combinations thereof. produced via a thermocouple. 6. The system of claim 1, wherein said energy transfer 26. The method of claim 25, wherein said electricity is used system is selected from the group consisting of a fuel cell, a for electrolysis of water. combustion unit, a thermocouple, and a heat transfer system. 27. A method comprising: 7. The system of claim 6, wherein said combustion unit a) providing a biomass and a population of at least one comprises a steam powered system. genus of a hyperthermophilic organism; 8. The system of claim 7, wherein said steam powered b) degrading said biomass in the presence of said popula system is a steam turbine or generator. tion of at least one genus of a hyperthermophilic organ 9. The system of claim 6, wherein said heat transfer system ism under conditions such that degradation products are comprises a heat pump. produced. 28. The method of claim 27, wherein said hyperthermo 10. The system of claim 6, wherein said energy transfer philic organisms are anaerobic hyperthermophilic organisms. system is a thermocouple and wherein said energy transfer 29. The method of claim 28, wherein said anaerobic hyper system further comprises an electrolysis system that coverts thermophilic organisms are selected from the group consist water into hydrogen and oxygen. ing of the genera Pyrococcus, Thermococcus, Acidianus, 11. The system of claim 1, wherein said biomass is selected Palaeococcus, Thermoplasma, Pyrobaculum, Pyrolobus, from the group consisting of sewage, agricultural waste prod Methanobacterium, and Thermotoga, and combination ucts, brewery grain by-products, food waste, organic industry thereof. waste, forestry waste, crops, grass, seaweed, plankton, algae, 30. The method of claim 27, wherein said hyperthermo fish, fish waste, and combinations thereof. philic organisms are aerobic hyperthermophilic organisms. 12. The system of claim 1, wherein said biomass is Supple 31. The method of claim 27, wherein said aerobic hyper mented with a cell culture media component selected from thermophilic organisms are selected from the group consist the group consisting of a mineral source, Vitamins, amino ing of Aeropyrum permix, Sulfolobus solfataricus, Metal acids, an energy source, and a microorganism extract. losphaera sedula, Sulfobus tokodaii, Thermoplasma 13. The system of claim 12, wherein said mineral source is acidophilum and Thermoplasma volcanium, and combina selected from the group consisting of NaCl, MgSO, MgCl, tions thereof. CaCl, KC1, NaBr, HBO, SrC1, MnSO4, FeSO, CoSO, 32. The method of claim 27, wherein said biomass is ZnSO, CuSO KAl(SO), NaMoOSO, (NHSO)2Ni selected from the group consisting of sewage, agricultural (SO), NaWO and NaSeO and combinations thereof. waste products, brewery grain by-products, food waste, 14. The system of claim 12, wherein said microorganism organic industry waste, forestry waste, crops, grass, Seaweed, extract is a yeast extract. plankton, algae, fish, fish waste, and combinations thereof. 15. The system of claim 12, wherein said energy source is 33. The method of claim 27, wherein said degradation starch. products are selected from the group consisting of hydrogen, 16. A method comprising: methane and ethanol. a) providing a biomass and a population of at least one 34. The method of claim 27, further comprising the step of genus of a hyperthermophilic organism; converting said degradation products into energy. b) fermenting said biomass in the presence of said popula 35. The method of claim 33, further comprising using said tion of at least one genus of a hyperthermophilic organ hydrogen in a fuel cell. 36. The method of claim 33, further comprising using said ism under conditions such that heat is produced; methane or ethanol in a combustion unit. c) using said heat to produce electricity or heat a liquid. 37. A method for reducing carbon dioxide emissions com 17. The method of claim 16, wherein said hyperthermo prising: philic organisms are anaerobic hyperthermophilic organisms. a) providing a biomass and a population of at least one 18. The method of claim 17, wherein said anaerobic hyper genus of a hyperthermophilic organism; thermophilic organisms are selected from the group consist b) anaerobically degrading said biomass in the presence of ing of the genera Pyrococcus, Thermococcus, Acidianus, said population of at least one genus of a hyperthermo Palaeococcus, Thermoplasma, Pyrobaculum, Pyrolobus, philic organism to produce Substrates for energy produc Pyrodictium, Methanotehrmus, Methanopyrus, and Fervido tion; bacterium Thermotoga, and combination thereof. c) producing energy from said Substrates, wherein carbon 19. The method of claim 16, wherein said hyperthermo dioxide emissions are reduced as compared to aerobic philic organisms are aerobic hyperthermophilic organisms. degradation of said biomass materials. 20. The method of claim 19, wherein said aerobic hyper 38. A method for generating carbon credits comprising: thermophilic organisms are selected from the group consist a) providing a biomass and a population of at least one ing of Aeropyrum permix, Sulfolobus solfataricus, Metal genus of a hyperthermophilic organism; losphaera sedula, Sulfobus tokodai, Thermoplasma b) anaerobically degrading said biomass in the presence of acidophilum and Thermoplasma volcanium, and combina said population of at least one genus of a hyperthermo tions thereof. philic organism to produce Substrates for energy produc 21. The method of claim 16, wherein said biomass is tion, selected from the group consisting of sewage, agricultural c) producing energy from said Substrates under conditions waste products, brewery grain by-products, food waste, Such that carbon credits are generated. organic industry waste, forestry waste, crops, grass, Seaweed, plankton, algae, fish, fish waste, and combinations thereof. c c c c c