Disclosed are trans-enoyl CoA reductase (TER) enzymes and nucleic acids encoding them. In some cases, the TER enzymes are non-natural, engineered trans-enoyl CoA reductase. TER enzymes were shown to catalyse the conversion of 5-carboxy-2-pentenoyl-CoA into adipyl-CoA for improved adipate production and the conversion of crotonyl-CoA into 6-aminocaproate. The enzymes can be used in biosynthetic methods and engineered microorganisms that enhance or improve the biosynthesis of 6-aminocaproate, hexamethylenediamine, caproic acid, caprolactone, or caprolactam. The engineered microorganisms include exogenous TER and in some cases engineered TER.

Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as adipate, 6-aminocaproate, hexamethylenediamine or caprolactam. Also provided herein are methods for using such an organism to produce adipate, 6-aminocaproate, hexamethylenediamine or caprolactam.

A method for oxidizing an alkyl, including a) contacting the alkyl with an aqueous solution comprising a microorganism where the microorganism has a reduced fatty acid degradation capacity compared to its wild type, wherein the fatty acid degradation capacity is reduced by deletion, inhibition or inactivation of a gene encoding an enzyme involved in the β-oxidation pathway; and the microorganism expresses a recombinant alkane oxidase, and b) contacting the aqueous solution from a) with a water-immiscible organic solvent.

The present invention provides at least two modified gene sequences, Sequence 1 comprising of nucleotide sequence of SEQ. ID no.1, and Sequence 2 comprising of nucleotide sequence of SEQ. ID no.2, encoding the enzyme choline oxidase wherein the gene sequences have been obtained by modifying the codA gene (Accession no. X84895) encoding choline oxidase from Arthrobacter globiformis, and a method to enzymatically produce betaine using choline oxidases encoded by Sequence 1, and Sequence 2, wherein the enzymatically produced betaine has minimal undesired trimethylamine contamination.

The present disclosure concerns a method for producing peptides such as glutathione and a microorganism that can be used for such method. One or more embodiments of the first aspect of the present disclosure concern a method for producing peptides such as glutathione comprising culturing a prokaryotic microbial strain in which the expression levels of one or more genes selected from among the gshA gene, the gshB gene, and the gshF gene are enhanced, compared with the expression levels thereof in the wild-type strain thereof in a medium in which the total concentration of cysteine and cystine is 0.5 g/l or lower. The second aspect of the present disclosure concerns a microorganism comprising disruptions of the γ-glutamyltransferase gene and the glutathione reductase gene and exhibiting the enhanced expression levels of the gshA gene and the gshB or gshF gene.

The present disclosure relates to a microorganism for producing a mycosporine-like amino acid, and a method for producing a mycosporine-like amino acid using the microorganism. The microorganism of the present disclosure shows an improved ability for producing a mycosporine-like amino acid and thus can be effectively used in the production of a mycosporine-like amino acid.

The invention provides a non-naturally occurring microbial organism having an adipate, 6-aminocaproic acid or caprolactam pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective adipate, 6-aminocaproic acid or caprolactam pathway. The invention additionally provides a method for producing adipate, 6-aminocaproic acid or caprolactam. The method can include culturing an adipate, 6-aminocaproic acid or caprolactam producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an adipate, 6-aminocaproic acid or caprolactam pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce adipate, 6-aminocaproic acid or caprolactam.

The present disclosure generally relates to devices, compositions and methods for designing and producing nucleic acid molecules and the production of encoded proteins using these nucleic acid molecules. In some aspect, the disclosure relates to automation for the in vitro generation of coding DNA molecules, the in vitro transcription of these DNA molecules to generate protein coding RNA molecules, and the in vitro translation of these protein coding RNA molecules to produce proteins.

The present disclosure provides methods for preparing β-substituted tryptophan compounds. The methods include: combining i) an unsubstituted indole or a substituted indole, ii) a β-substituted serine, and iii) a tryptophan synthase β-subunit (i.e., a TrpB); and maintaining the resulting mixture under conditions sufficient to form the β-substituted tryptophan. The TrpB contains at least one amino acid mutation which promotes formation of an amino-acrylate intermediate. New TrpB variants and new β-substituted tryptophan analogs are also described.

The invention provides a non-naturally occurring microbial organism having a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The invention additionally provides a method for producing 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid. The method can include culturing a 6-aminocaproic acid, caprolactam or hexametheylenediamine producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid.