Method for the production of glutaconate

摘要

The present invention relates to a novel method for the biocatalytic production of unsaturated dicarboxylic acids by cultivating a recombinant microorganism co-expressing a glutaconate CoA-transferase and a 2-hydroxyglutaryl-CoA dehydratase system. The present invention also relates to corresponding recombinant hosts, recombinant vectors, expression cassettes and nucleic acids suitable for preparing such hosts as well as a method of preparing polyamide or polyester copolymers making use of said dicarboxylic acids as obtained by said biocatalytic production method.

主权项

1. A biocatalytic method for the production of an unsaturated dicarboxylic acid compound of formula I
HOOC—CH═CH—X—COOH  (I) wherein X represents a linear or branched, optionally unsaturated, optionally substituted hydrocarbon group, wherein X is selected from (CH2)n, with n being an integer from 1 to 4, CH═CH, CH2-C(═O), or CH═C(OH), wherein said method comprises: converting a 2-hydroxy-substituted dicarboxylic acid compound of formula III
HOOC—C(OH)H—CH2-X—COOH  (III) wherein X is as defined above; in a recombinant microorganism co-expressing a glutaconate CoA-transferase and a 2-hydroxyglutaryl-CoA dehydratase, optionally in the presence of a coenzyme A source so that said compound of formula I is formed; and optionally isolating said compound of formula I in the form of a substantially pure stereoisomer or as a mixture of stereoisomers, wherein said glutaconate CoA-transferase comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 4 and/or 6, SEQ ID NO: 18 and/or 20, or SEQ ID NO: 22 and/or 24, and wherein said 2-hydroxyglutaryl-CoA dehydratase comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8 and/or 10, SEQ ID NO: 26 and/or 28, or SEQ ID NO: 30, 32 and/or 34.