The present disclosure provides methods for utilizing genetically modified microbes to co-produce 3-hydroxypropionic acid (3-HP) and acetyl-CoA, and derivatives thereof from malonate semialdehyde as a common single intermediate. The disclosure further provides modified microbe that co-produce the 3-HP and acetyl-CoA derivatives from malonate semialdehyde.

Microorganisms are genetically engineered to produce various chemicals for industrial use. The microorganisms are carboxydotrophic acetogens. The microorganisms produce acetyl-CoA using the Wood-Ljungdahl Pathway for fixing CO/CO.sub.2. A reverse beta-oxidation pathway cycle from a microorganism that contains such a group of enzymes is introduced. Additionally, primers and extenders, and/or genes encoding for enzymes that generate primers and extenders may also be introduced. Product synthesis can be effected by improved promoters or enzyme designs that are catalytically more efficient. Similarly, product synthesis may also be improved by deleting competing reactions.

Disclosed herein is a novel, cofactor balanced, cell-free biocatalysis pathway to make diacids from formaldehyde or methanol that does not use ATP and produces no CO.sub.2. In another embodiment, disclosed herein is a novel C5/C6 (hydrolysate) utilization pathway that interplays with the cell making diacids from formaldehyde or methanol that does not use ATP, produces no CO.sub.2, and replaces ATP with (cheap) polyphosphate (no other cofactors needed). Using methods and compositions disclosed herein, no CO.sub.2 is produced and the pathways disclosed herein are cell-free. The combination of pathways, feedstocks and products disclosed herein is novel.

Disclosed herein is a novel, cofactor balanced, cell-free biocatalysis pathway to make diacids from formaldehyde or methanol that does not use ATP and produces no CO.sub.2. In another embodiment, disclosed herein is a novel C5/C6 (hydrolysate) utilization pathway that interplays with the cell making diacids from formaldehyde or methanol that does not use ATP, produces no CO.sub.2, and replaces ATP with (cheap) polyphosphate (no other cofactors needed). Using methods and compositions disclosed herein, no CO.sub.2 is produced and the pathways disclosed herein are cell-free. The combination of pathways, feedstocks and products disclosed herein is novel.

A system optionally including a carbon oxide reactor. A method for carbon oxide reactor control, optionally including selecting carbon oxide reactor aspects based on a desired output composition, running a carbon oxide reactor under controlled process conditions to produce a desired output composition, and/or altering the process conditions to alter the output composition.

The invention relates to a method for producing salts of butyric acid, lactic acid, propionic acid, valeric acid, acetic acid, glycolic acid, sorbic acid, fumaric acid, formic acid, malic acid, tartaric acid, citric acid, derivatives of said organic acids and mixtures thereof, comprising at least the successive steps of: •i) culturing a microorganism in an appropriate culture medium comprising a carbon source and a nitrogen source, during a sufficient time to obtain a fermentation broth comprising the organic acid(s); •ii) pre-treating said fermentation broth comprising at least clarification of the fermentation broth and separation of organic acids by evaporation of the clarified fermentation broth and then condensation of the volatile fraction containing organic acids (CVAF); •v) adding inorganic bases to said CVAF •vii) eliminating the remaining water of the CVAF, and recovering the organic acid salts.

The invention relates to a method for producing salts of butyric acid, lactic acid, propionic acid, valeric acid, acetic acid, glycolic acid, sorbic acid, fumaric acid, formic acid, malic acid, tartaric acid, citric acid, derivatives of said organic acids and mixtures thereof, comprising at least the successive steps of: •i) culturing a microorganism in an appropriate culture medium comprising a carbon source and a nitrogen source, during a sufficient time to obtain a fermentation broth comprising the organic acid(s); •ii) pre-treating said fermentation broth comprising at least clarification of the fermentation broth and separation of organic acids by evaporation of the clarified fermentation broth and then condensation of the volatile fraction containing organic acids (CVAF); •v) adding inorganic bases to said CVAF •vii) eliminating the remaining water of the CVAF, and recovering the organic acid salts.

Provided is a method for construction of recombinant bacteria for producing 3-hydroxypropionic acid. The method includes: knocking out fadR, fabF and fabH genes of recipient bacteria, introducing acc genes or gene clusters, alKL and Mcr genes, and enhancing the expression of fadL, fadD, sthA genes and atoSC gene clusters in the recipient bacteria. Also provided is a method for producing 3-hydroxypropionic acid by using the recombinant bacteria.

Provided is a method for construction of recombinant bacteria for producing 3-hydroxypropionic acid. The method includes: knocking out fadR, fabF and fabH genes of recipient bacteria, introducing acc genes or gene clusters, alKL and Mcr genes, and enhancing the expression of fadL, fadD, sthA genes and atoSC gene clusters in the recipient bacteria. Also provided is a method for producing 3-hydroxypropionic acid by using the recombinant bacteria.

The disclosure relates to engineered enone reductase polypeptides having improved properties, polynucleotides encoding the engineered polypeptides, related vectors, host cells, and methods for making the engineered enone reductase polypeptides. The disclosure also provides methods of using the engineered enone reductase polypeptides for chemical transformations.