Abstract: Disclosed are methods for engineering bacteria, for example, Thermoanaerobacterium saccharolyticum, that convert biomass to ethanol at high yield by deleting a single gene. Deletion of subunit A or subunit B of the hfs hydrogenase, but not deletion of subunit C or subunit D, results in an increase in ethanol yield.

Abstract: A system and method for converting biomass with no chemical pretreatment is disclosed. Combination of a microbial system and the use of mechanical disruption during fermentation may help achieve high conversion rate without the extra cost and undesirable by-products typically associated with the pretreatment process.

Abstract: Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2. The organisms may be utilized for example in thermophilic SSF and SSCF reactions performed at temperatures that are optimal for cellulase activity to produce near theoretical ethanol yields without expressing pyruvate decarboxylase.

Abstract: It is disclosed here engineered cellulolytic microorganisms capable of producing ethanol from lignocellulosic feedstock with high yield. Multiple genes in Thermoanaerobacterium saccharolyticum that are involved in the pyruvate to ethanol pathway are disclosed which may be transferred into C. thermocellum or other natively cellulolytic microorganisms.

Abstract: Disclosed are methods for engineering bacteria, for example, Thermoanaerobacterium saccharolyticum, that convert biomass to ethanol at high yield by deleting a single gene. Deletion of subunit A or subunit B of the hfs hydrogenase, but not deletion of subunit C or subunit D, results in an increase in ethanol yield.

Abstract: A system and method for converting biomass with no chemical pretreatment is disclosed. Combination of a microbial system and the use of mechanical disruption during fermentation may help achieve high conversion rate without the extra cost and undesirable by-products typically associated with the pretreatment process.

Abstract: A new and improved biomass conversion system is disclosed using high-temperature flow-though pretreatment and a nanoporous membrane to provide more digestible biomass for subsequent conversion to biofuels.

Abstract: A process for the treatment of biomass to render structural carbohydrates more accessible and/or digestible using concentrated ammonium hydroxide with or without anhydrous ammonia addition, is described. The process preferably uses steam to strip ammonia from the biomass for recycling. The process yields of monosaccharides from the structural carbohydrates are good, particularly as measured by the enzymatic hydrolysis of the structural carbohydrates. The monosaccharides are used as animal feeds and energy sources for ethanol production.

Abstract: A new species of an anaerobic thermophilic cellulolytic and xylano lytic bacterium is disclosed. One particular strain of this new species has been deposited with the ATCC under Deposit No. PTA-10114. It is also provided a method for isolating, culturing and utilizing this novel bacterium for the conversion of biomass to bioconversion products, such as ethanol.

Abstract: Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2. The organisms may be utilized for example in thermophilic SSF and SSCF reactions performed at temperatures that are optimal for cellulase activity to produce near theoretical ethanol yields without expressing pyruvate decarboxylase.

Abstract: Methods for obtaining cellulolytic microbes with high growth rates are disclosed. For example, C. thermocellum colonies with growth rates higher than 0.17 hr? 1 have been obtained by the present methods. In realizing higher growth rates, better bioprocessing efficiency can be achieved resulting in increased economy.

Abstract: Recombinant yeast strains that saccharify, ferment and grow on insoluble and crystalline forms of cellulose are disclosed herein. The yeast strains express tethered cellulases including cellobiohydrolase, endoglucanase and ?-glucosidase. The recombinant organisms are particularly suited for consolidated bioprocessing.

Abstract: Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2.

Abstract: Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2.

Abstract: Mutant thermophilic organisms that consume a variety of biomass derived substrates are disclosed herein. Strains of Thermoanaerobacterium saccharolyticum with acetate kinase and phosphotransacetylase expression eliminated are disclosed herein. Further, strain ALK1 has been engineered by site directed homologous recombination to knockout both acetic acid and lactic acid production. Continuous culture involving a substrate concentration challenge lead to evolution of ALK1, and formation of a more robust strain designated ALK2. The organisms may be utilized for example in thermophilic SSF and SSCF reactions performed at temperatures that are optimal for cellulase activity to produce near theoretical ethanol yields without expressing pyruvate decarboxylase.

Abstract: A process for the treatment of biomass to render structural carbohydrates more accessible and/or digestible using concentrated ammonium hydroxide with or without anhydrous ammonia addition, is described. The process preferably uses steam to strip ammonia from the biomass for recycling. The process yields of monosaccharides from the structural carbohydrates are good, particularly as measured by the enzymatic hydrolysis of the structural carbohydrates. The monosaccharides are used as animal feeds and energy sources for ethanol production.

Abstract: A method for separating saccharide components and lignin fractions from a concentrated acid treated lignocellulosic biomass is disclosed. The method involves precipitating the saccharide components by adding an organic solvent to the biomass slurry. The acid may then be recovered, for example, by filtration or by countercurrent washing and the organic solvent may be flashed and recycled. During acid recovery and organic recovery steps, two main lignocellulose components (hemicellulose and lignin) as well as minor components such as acetic acid are separated as well. The method decreases the amount of cellulase required for hydrolysis, increases hydrolysis rates, reduces formation of inhibitor molecules, increase sugar yields, produces high value by-products such as high quality lignin and hemicellulose, and decreases energy and equipment costs.

Abstract: A method and apparatus for electroporation includes placing a mixture of bacterial suspension and transforming DNA into an electroporation cuvette. The resulting sample is subjected through a current-limiting device to a complex 5 waveform including a burst of high-voltage radio-frequency current, which in some embodiments is superimposed on a biphasic high-voltage DC pulse, and in other embodiments on a high-voltage lower-frequency AC burst. The total waveform has at least an initial portion greater than eleven thousand volts per centimeter of electrode spacing, and a later portion in some embodiments is reduced to less than thirty percent 10 of magnitude of the initial portion. Transformed bacteria are selected by culture in selective medium in an embodiment. The high-voltage radio-frequency current is between 3 and 125 MHz, and in an embodiment is 24 MHz.

Abstract: Cellodextrins are made by cellulose hydrolysis using mixed concentrated hydrochloric acid and sulfuric acid. Cellulose is rapidly and completely dissolved to produce a high cellodextrin yield. Acetone is used as an efficient organic solvent for precipitating cellodextrins from the mixed acid hydrolyzate. Cellodextrins are resuspended in water and separated with high productivity using ion exchange.

Abstract: An improved continuous process for producing ethanol from cellulosic materials employs an intermittently agitated, perpetually fed, solids retaining reactor vessel. Cellulosic substrate, catalysts and fermentation agents are introduced into a reaction vessel to form a slurry. The slurry is agitated for a first selected time interval under conditions sufficient to initiate and maintain a fermentation reaction and then allowed to settle during a second selected time interval. At the end of the second selected time interval an ethanol-containing effluent is removed from the vessel. Thereafter, additional cellulosic material, catalysts and fermentation agents are added to the reactor vessel. This operating cycle is repeated virtually perpetually to effect the ethanol production. It can be implemented using a single bioreactor or a cascade of bioreactors.