Preparation of 6-aminocaproic acid from 5-formyl valeric acid

Abstract

The invention relates to a method for preparing 6-aminocaproic acid (hereinafter also referred to as ‘6-ACA’) using a biocatalyst. The invention further relates to a method for preparing ε-caprolactam (hereafter referred to as ‘caprolactam’) by cyclising such 6-ACA. The invention further relates to a host cell, a micro-organism, or a polynucleotide which may be used in the preparation of 6-ACA or caprolactam.

1 . (canceled) 
     
     
         2 . Method for preparing 6-aminocaproic acid, wherein the 6-aminocaproic acid is prepared from 5-formylpentanoate, using a recombinant host cell comprising at least one biocatalyst, wherein the biocatalyst is an enzyme capable of catalysing a transamination and/or a reductive amination selected from the group of aminotransferases (E.C. 2.6.1) and amino acid dehydrogenases (E.C.1.4.1). 
     
     
         3 - 4 . (canceled) 
     
     
         5 . Method according to  claim 2 , wherein the aminotransferase or amino acid dehydrogenase is selected from the group of β-aminoisobutyrate:α-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). 
     
     
         6 . Method according to  claim 5 , wherein the enzyme is selected from the group of enzymes capable of catalysing a transamination and/or a reductive amination from an organism selected from the group of  Vibrio; Pseudomonas; Bacillus; Mercurialis; Asplenium; Ceratonia ; mammals;  Neurospora; Escherichia; Thermus; Saccharomyces; Brevibacterium; Corynebacterium; Proteus; Agrobacterium; Geobacillus; Acinetobacter; Ralstonia; Salmonella; Rhodobacter  and  Staphylococcus , in particular from an organism selected from the group of  Bacillus subtilis, Bacillus weihenstephanensis, Rhodobacter sphaeroides, Staphylococcus aureus, Legionella pneumophila, Nitrosomonas europaea, Neisseria gonorrhoeae, Pseudomonas syringae, Rhodopseudomonas palustris, Vibrio fluvialis  and  Pseudomonas aeruginosa.    
     
     
         7 . Method according to  claim 6 , wherein an aminotransferase is used comprising an amino acid sequence according to Sequence ID 2, Sequence ID 5, Sequence ID 8, Sequence ID 12, Sequence ID 15, Sequence ID 17, Sequence ID 19, Sequence ID 21, Sequence ID 23, Sequence ID 25, Sequence ID 27, Sequence ID 29, Sequence ID 65, Sequence ID 67, Sequence ID 69 or a homologue of any of these sequences. 
     
     
         8 - 15 . (canceled) 
     
     
         16 . Method for preparing caprolactam, comprising cyclising the 6-aminocaproic acid prepared by the method of  claim 2  in the presence of superheated steam, thereby forming caprolactam. 
     
     
         17 . A recombinant host cell comprising a nucleic acid sequence encoding an enzyme with 5-formylpentanoate aminotransferase activity. 
     
     
         18 . A recombinant host cell according to  claim 17 , comprising a nucleic acid sequence encoding an enzyme with 5-formylpentanoate aminotransferase comprising an amino acid sequence according to Sequence ID 2, Sequence ID 5, Sequence ID 8, Sequence ID 65 Sequence ID 67, Sequence ID 69 or a homologue thereof. 
     
     
         19 - 22 . (canceled) 
     
     
         23 . A recombinant host cell according to  claim 18 , wherein the host cell is selected from the group of  Aspergillus, Penicillium, Saccharomyces, Kluyveromyces, Pichia, Candida, Hansenula, Bacillus, Corynebacterium , and  Escherichia.    
     
     
         24 . A micro-organism according to  claim 23  comprising DNA containing a nucleic acid sequence selected from the group of sequences represented by any sequence selected from the group of Sequence ID 1, Sequence ID 3, Sequence ID 4, Sequence ID 6, Sequence ID 7, Sequence ID 64, Sequence ID 66, Sequence ID 68 and functional analogues thereof. 
     
     
         25 . (canceled)