Methods of producing 6-carbon chemicals via methyl-ester shielded carbon chain elongation

摘要

This document describes biochemical pathways for producing adipic acid, 6-aminohexanoic acid, 6-hydroxhexanoic acid, hexamethylenediamine, caprolactam, or 1,6-hexanediol by forming one or two terminal functional groups, comprised of carboxyl, amine or hydroxyl group, in a C6 aliphatic backbone substrate. These pathways, metabolic engineering and cultivation strategies described herein rely on the enzymes or homologs accepting methyl ester shielded dicarboxylic acid substrates.

主权项

1. A method for biosynthesizing a product selected from the group consisting of adipic acid, 6-aminohexanoate, 6-hydroxhexanoate, hexamethylenediamine, caprolactam and 1,6-hexanediol, said method comprising, in a recombinant host, enzymatically synthesizing a six carbon chain aliphatic backbone from oxalyl-CoA and either (i) acetyl-CoA or malonyl-CoA via two cycles of methyl ester shielded carbon chain elongation or (ii) malonyl-[acp] via two cycles of methyl-ester shielded carbon chain elongation, and enzymatically forming two terminal functional groups selected from the group consisting of carboxyl, amine, and hydroxyl groups in said backbone, thereby forming the product, wherein:
a polypeptide having the activity of a malonyl-[acp] O-methyltransferase converts oxalyl-CoA to oxalyl-CoA methyl ester, and each of said two cycles of carbon chain elongation comprises using a polypeptide having the activity of (i) a β-ketoacyl-[acp] synthase or a β-ketothiolase, (ii) a 3-oxoacyl-[acp] reductase, an acetoacetyl-CoA reductase, a 3-hydroxyacyl-CoA dehydrogenase or a 3-hydroxybutyryl-CoA dehydrogenase, (iii) an enoyl-CoA hydratase or a 3-hydroxyacyl-[acp] dehydratase, and (iv) an enoyl-[acp] reductase or a trans-2-enoyl-CoA reductase to produce adipyl-CoA methyl ester or adipyl-[acp] methyl ester.