The production of ω-hydroxy palmitic acid using fatty acid metabolism and cofactor optimization in Escherichia coli

摘要: Hydroxylated fatty acids (HFAs) are used as important precursors for bulk and fine chemicals in the chemical industry. Here, to overproduce long-chain (C16–C18) hydroxy acid, their biosynthetic pathways including thioesterase (Lreu_0335) from Lactobacillus reuteri DSM20016, β-hydroxyacyl-ACP dehydratase (fabZ) Escherichia coli, a P450 system (i.e., CYP153A Marinobacter aquaeolei VT8 camA/camB Pseudomonas putida ATCC17453) were overexpressed. Acyl-CoA synthase (fadD) involved acid degradation by β-oxidation was also deleted E. coli BW25113. The engineered FFA4 strain without could produce 503.0 mg/l of palmitic (C16) 508.4 stearic (C18) acids, which amounts ca. 1.6- 2.3-fold higher than those wild type. On other hand, HFA4 ω-hydroxylation 211.7 ω-hydroxy 42.1 ± 0.1 % generated indicating that hydroxylation reaction rate-determining step HFA production. For maximum production NADH, i.e., an essential cofactor reaction, overproduced integration NAD+-dependent formate dehydrogenase (FDH) Candida boidinii into chromosome deletion alcohol (ADH). Finally, NADH-level-optimized produced 610 (ω-HPA), almost threefold increase its yield compared same NADH overproduction.