Recombinant microorganisms exhibiting increased flux through a fermentation pathway
AssigneeLanzatech New Zealand Limited
InventorsMichael Koepke, Alexander Paul Mueller
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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.
Claims20
A method of producing a fermentation product, the method comprising:
- a) determining a rate-limiting pathway reaction in a fermentation pathway;
- b) identifying at least one enzyme, co-factor or both, which are involved in catalysing the rate-limiting pathway reaction;
- c) fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit at least one of:
- i) increased activity of the at least one enzyme of (b) or a functionally equivalent variant thereof, or ii) increased availability of the at least one co-factor of (b), when compared to a parental microorganism, to produce a fermentation product.
A method of producing a fermentation product, the method comprising fermenting a CO-comprising substrate with a recombinant carboxydotrophic Clostridia microorganism to produce a fermentation product, wherein the recombinant microorganism is adapted to exhibit at least one of:
- a) increased activity of at least one enzyme identified as being involved in catalysing a rate-limiting pathway reaction of a fermentation pathway, or a functionally equivalent variant thereof, when compared to a parental microorganism;
- or
- b) increased availability of at least one co-factor identified as being involved in catalysing a rate-limiting pathway reaction of a fermentation pathway, when compared to a parental microorganism.
A method of producing a recombinant carboxydotrophic Clostridia microorganism adapted to exhibit increased flux through a fermentation pathway relative to a parental microorganism, the method comprising:
- a) determining a rate limiting pathway reaction in the fermentation pathway;
- b) identifying at least one enzyme, co-factor or both, which are involved in catalysing the rate-limiting pathway reaction;
- c) transforming a parental microorganism to yield a recombinant microorganism adapted to exhibit at least one of i) increased activity of the at least one enzyme of (b) or a functionally equivalent variant thereof, or ii) increased availability of the at least one co-factor of (b), when compared to a parental microorganism;
- wherein the fermentation pathway is capable of producing at least one fermentation product from a substrate comprising CO.
A recombinant carboxydotrophic Clostridia microorganism adapted to exhibit at least one of
- a) increased activity of at least one enzyme or a functionally equivalent variant thereof when compared to a parental microorganism;
- b) increased availability of at least one co-factor when compared to a parental microorganism;
- or
- c) both a) and b);
- wherein the at least one enzyme or co-factor have been identified as being involved in catalysing a rate-limiting pathway reaction.
Dependencies are inferred from the claim text.