Abstract: The disclosure provides genetically engineered microorganisms and methods for improved biological production of ethylene glycol and precursors of ethylene glycol. The microorganism of the disclosure produces ethylene glycol or a precursor of ethylene glycol through one or more of 5,10-methylenetetrahydrofolate, oxaloacetate, citrate, malate, and glycine. The disclosure further provides compositions comprising ethylene glycol or polymers of ethylene glycol such as polyethylene terephthalate.

Abstract: The disclosure relates to the system comprising a combination of a primary bioreactor with a secondary bioreactor to convert a gaseous stream comprising CO and optionally H2, or CO2 and H2 to at least one target product. Bioreactors may be any combination of aerobic and anaerobic, batch and continuous. The bioreactors comprise fermenting microorganisms that may typically be bacterial or fungal.

Abstract: The disclosure relates to the combination of a primary fermentation that converts gas to acetate with a secondary fermentation that converts acetate to a target product. Preferably, the gas contains carbon dioxide, such that the disclosure enables the fixation of carbon dioxide into useful products. The fermentations may be any combination of aerobic and anaerobic, batch and continuous. The fermenting microorganisms may typically be bacterial or fungal.

Abstract: The disclosure provides genetically engineered microorganisms and methods for improved biological production of ethylene glycol and precursors of ethylene glycol. The microorganism of the disclosure produces ethylene glycol or a precursor of ethylene glycol through one or more of 5,10-methylenetetrahydrofolate, oxaloacetate, citrate, malate, and glycine. The disclosure further provides compositions comprising ethylene glycol or polymers of ethylene glycol such as polyethylene terephthalate.

Abstract: The disclosure relates to the combination of a primary fermentation that converts gas to acetate with a secondary fermentation that converts acetate to a target product. Preferably, the gas contains carbon dioxide, such that the disclosure enables the fixation of carbon dioxide into useful products. The fermentations may be any combination of aerobic and anaerobic, batch and continuous. The fermenting microorganisms may typically be bacterial or fungal.

Abstract: Disclosed are compositions, methods, and kits for performing cell-free RNA transcription and/or cell-free protein synthesis (CFPS). The disclosed compositions, methods, and kits include or utilize components prepared from a species of Clostridia such as cellular extracts from Clostridium autoethanogenum.

Abstract: The invention provides a recombinant, carboxydotrophic Clostridium bacterium that expresses one or more of pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1), acetolactate synthase (EC 2.2.1.6), and acetolactate decarboxylase (EC 4.1.1.5). The invention further provides a method of producing a fermentation product by fermenting the recombinant bacterium in the presence of a gaseous substrate comprising CO to produce one or more of ethanol, butanol, isopropanol, isobutanol, higher alcohols, butanediol, 2,3-butanediol, succinate, isoprenoids, fatty acids, biopolymers, and mixtures thereof.

Abstract: The invention relates to a genetically engineered bacterium having an enzyme that converts acetyl-CoA to acetoacetyl-CoA, an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and an enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate. The bacterium may also have enzymes to produce other downstream products, such as 3-hydroxybutyryaldehyde, and 1,3-butanediol. Typically, the bacterium is capable of producing these products from a gaseous substrate, such as syngas or an industrial waste gas.

Abstract: The invention relates to a genetically engineered bacterium having an enzyme that converts acetyl-CoA to acetoacetyl-CoA, an enzyme that converts acetoacetyl-CoA to 3-hydroxybutyryl-CoA, and an enzyme that converts 3-hydroxybutyryl-CoA to 3-hydroxybutyrate. The bacterium may also have enzymes to produce other downstream products, such as 3-hydroxybutyryaldehyde, and 1,3-butanediol. Typically, the bacterium is capable of producing these products from a gaseous substrate, such as syngas or an industrial waste gas.

Abstract: Carboxydotrophic acetogenic microorganisms do not produce MEK and/or 2-butanol. They lack the biosynthesis pathways to make these products. In addition, they produce the intermediate (R,R)-2,3-butanediol whereas the production of MEK and 2-butanol requires production of the intermediate (R,S)-2,3-butanediol. Nonetheless, the production of MEK and/or 2-butanol can be accomplished using recombinant microorganisms adapted to express or overexpress key enzymes in the MEK and/or 2-butanol biosynthesis pathways. Such microorganisms, such as the carboxydotrophic acetogen Clostridium autoethanogenum, can ferment substrates comprising CO. The overall scheme involves the production of 2-butanol from (R,S)-2,3-butanediol and the conversion of (R)-acetoin to (S)-2,3-butanediol. These steps are involved in the production of both MEK and 2-butanol. Such fermentation methods offer a means of using carbon monoxide from industrial processes which would otherwise be released into the atmosphere and pollute the environment.

Abstract: A methods for altering the metabolite profile of a fermentation, by increasing flux through acetolactate. The methods comprises increasing production of one or more products derived from acetolactate. Further provided is a method for increasing the production of 2,3-butandiol by microbial fermentation of gaseous substrates, the method comprising providing a compound which inhibits one or more enzymes which convert acetolactate to branched chain amino acids to the fermentation. The present invention further provides methods for increasing the production of 2,3-butandiol relative to other fermentation products such as ethanol and acetic acid.

Abstract: The present provides selection markers, methods, nucleic acids, and vectors of use in the preparation of recombinant Clostridium spp.

Abstract: The invention provides a recombinant carboxydotrophic Clostridia microorganism with increased overall utilization of NADPH relative to a parent microorganism. Further, the invention provides a method of producing a recombinant carboxydotrophic Clostridia microorganism which exhibits increased NADPH utilization relative to a parental microorganism. In particular, the invention relates to increasing the overall utilization of NADPH in a recombinant carboxydotrophic Clostridia microorganism in order to increase the production of at least one fermentation product by the microorganism.

Abstract: The invention provides a recombinant, carboxydotrophic Clostridium bacterium that expresses one or more of pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1), acetolactate synthase (EC 2.2.1.6), and acetolactate decarboxylase (EC 4.1.1.5). The invention further provides a method of producing a fermentation product by fermenting the recombinant bacterium in the presence of a gaseous substrate comprising CO to produce one or more of ethanol, butanol, isopropanol, isobutanol, higher alcohols, butanediol, 2,3-butanediol, succinate, isoprenoids, fatty acids, biopolymers, and mixtures thereof.

Abstract: A stereospecific enzyme in C. autoethanogenum permits the conversion of racemic propanediol to acetone and/or propionaldehyde. Entantiomeric starting materials lead to different products. If desired, the products may be reduced to form alcohols. The reaction can be performed in various host cells, so that various materials may be used as carbon and/or energy sources.

Abstract: The invention provides a recombinant, acetogenic, carboxydotrophic bacterium that lacks secondary alcohol dehydrogenase or comprises an inactivated secondary alcohol dehydrogenase. The inactivated secondary alcohol dehydrogenase may be encoded by a secondary alcohol dehydrogenase gene comprising an inactivating mutation that reduces the ability of the bacterium to convert acetone to isopropanol and to convert methyl ethyl ketone to 2-butanol. Since a bacterium that lacks secondary alcohol dehydrogenase or comprises an inactivated secondary alcohol dehydrogenase accumulates carbonyl-containing compounds, the invention also provides a method of producing carbonyl-containing compounds, such as acetone and methyl ethyl ketone.

Abstract: A methods for altering the metabolite profile of a fermentation, by increasing flux through acetolactate. The methods comprises increasing production of one or more products derived from acetolactate. Further provided is a method for increasing the production of 2,3-butandiol by microbial fermentation of gaseous substrates, the method comprising providing a compound which inhibits one or more enzymes which convert acetolactate to branched chain amino acids to the fermentation. The present invention further provides methods for increasing the production of 2,3-butandiol relative to other fermentation products such as ethanol and acetic acid.

Abstract: The invention provides methods of increasing the production of fermentation products by increasing flux through a fermentation pathway by optimising enzymatic reactions. In particular, the invention relates to identifying enzymes and/or co-factors involved in metabolic bottlenecks in fermentation pathways, and fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit increased activity of the one or more of said enzymes, or increased availability of the one or more of said co-factors, when compared to a parental microorganism.

Abstract: The present provides selection markers, methods, nucleic acids, and vectors of use in the preparation of recombinant Clostridium spp.

Abstract: The invention provides a recombinant carboxydotrophic Clostridia microorganism with increased overall utilization of NADPH relative to a parent microorganism. Further, the invention provides a method of producing a recombinant carboxydotrophic Clostridia microorganism which exhibits increased NADPH utilization relative to a parental microorganism. In particular, the invention relates to increasing the overall utilization of NADPH in a recombinant carboxydotrophic Clostridia microorganism in order to increase the production of at least one fermentation product by the microorganism.