Microorganisms and methods for the biosynthesis of adipate, hexamethylenediamine and 6-aminocaproic acid

Method according to claim 2, wherein the aminotransferase or amino acid dehydrogenase is selected from the group of β-aminoisobutyrate:nα-ketoglutarate aminotransferases, β-alanine aminotransferases, aspartate aminotransferases, 4-amino-butyrate aminotransferases (EC 2.6.1.19), L-lysine 6-aminotransferase (EC 2.6.1.36), 2-aminoadipate aminotransferases (EC 2.6.1.39), 5-aminovalerate aminotransferases (EC 2.6.1.48), 2-aminohexanoate aminotransferases (EC 2.6.1.67), lysine:pyruvate 6-aminotransferases (EC 2.6.1.71), and lysine-6-dehydrogenases (EC 1.4.1.18).

Method according to claim 2, wherein an aminotransferase is used comprising an amino acid sequence according to:n(i) an enzyme from Vibrio fluvialis, Bacillus weihenstephanensis, Pseudomonas aeruginosa, Bacillus subtilis, or Pseudomonas aeruginosa that catalyses the conversion of adipate semialdehyde to 6-aminocaproic acid;(ii) an enzyme from Vibrio fluvialis, Pseudomonas aeruginosa, Pseudomonas syringae, Bacillus subtilis, Rhodobaeter sphaeroides, Legionella pneumophila, Nitrosomonas europaea, Neisseria gonorrhoeae, Pseudomonas aeruginosa, or Rhodopseudomonas palustris that catalyses the conversion of 2-OHD to 2-AHD; or(iii) a gene product of gabT from Escherichia coli, puuE from Escherichia coli, abat from Mus musculus, gabT from Pseudomonas fluorescens, or abat from Sus scrofa.

Method according to claim 6, wherein the enzyme capable of catalysing the decarboxylation is a decarboxylase (E.C. 4.1.1).

Method according to claim 6, wherein the enzyme capable of catalysing the decarboxylation is enzyme from an organism or part thereof selected from the group of Cucurbitaceae; Saccharomyces; Candida; Hansenula; Kluyveromyces; Rhizopus; Neurospora; Zymomonas; Escherichia; Mycobacterium; Clostridium; Lactobacillus; Streptococcus; Pseudomonas and Lactococcus.

Method according to claim 6, wherein the enzyme capable of catalysing the decarboxylation comprises an amino acid sequence according to:n(i) an enzyme from Escherichia coli, Saccharomyces cerevisiae, Zymomonas mobilis, Lactococcus lactis or Mycobacterium tuberculosis that catalyses the conversion of 2-OHD to adipate semialdehyde or 2-AHD to 6-aminocaproic acid; or(ii) a gene product of pdc from Zymomonas mobilis, pdc l from Saccharomyces cerevisiae, pdc from Acetobacter pasteurians, pdc l from Kluyveromyces lactis, mdIC from Pseudomonas putida, mdIC from Pseudomonas aeruginosa, dpgB from Pseudomonas stutzeri, ilvB-1 from Pseudomonas fluorescens, kgd from Mycobacterium tuberculosis, kgd from Bradyrhizobium japonicum, kgd from Mesorhizobium loti, kdcA from Lactococcus lactis, BCKDHB from Homo sapiens, BCKDHA from Homo sapiens, BCKDHB from Bos taurus, BCKDHA from Bos taurus, panD from Escherichia coli K12, panD from Corynebacterium glutamicum or panD from Mycobacterium tuberculosis.

Method according to claim 6, wherein 2-OHD is biocatalytically converted into adipate semialdehyde in the presence of a biocatalyst capable of catalysing the decarboxylation of an a-keto acid, and adipate semialdehyde is biocatalytically converted into 6-aminocaproic acid in the presence of at least one amino donor and at least one biocatalyst capable of catalysing a transamination and/or a reductive amination of adipate semialdehyde.

Method according to claim 15, wherein said biocatalyst for converting pyruvate and succinic semialdehyde to 2-OHD comprises an enzyme capable of catalysing the condensation of pyruvate and succinic semialdehyde to 4-hydroxy-2-oxo-heptane-1,7-dioate (HOHD).

Method according to claim 15, wherein said biocatalyst for converting pyruvate and succinic semialdehyde to 2-OHD comprises an enzyme capable of catalysing the dehydration of 4-hydroxy-2-oxo-heptane-1,7-dioate (HOHD) to 2-oxo-hept-4-ene-1,7-dioate (OHED).

Method according to claim 15, wherein said biocatalyst for converting pyruvate and succinic semialdehyde to 2-OHD comprises an enzyme capable of catalysing the reduction of 2-oxo-hept-4-ene-1,7-dioate (OHED) to 2-OHD.

Method according to claim 15, wherein said biocatalyst for converting pyruvate and succinic semialdehyde to 2-OHD comprises an enzyme capable of catalysing the condensation of pyruvate and succinic semialdehyde to 4-hydroxy-2-oxo-heptane-1,7-dioate (HOHD), an enzyme capable of catalysing the dehydration of HOHD to 2-oxo-hept-4-ene-1,7-dioate (OHED), and an enzyme capable of catalysing the reduction of OHED to 2-OHD.