caa a22 4500 605498415 CHVBK 20210128100546.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6630-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6630-1 The production of ω-hydroxy palmitic acid using fatty acid metabolism and cofactor optimization in Escherichia coli [Elektronische Daten] [Changmin Sung, Eunok Jung, Kwon-Young Choi, Jin-hyung Bae, Minsuk Kim, Joonwon Kim, Eun-Jung Kim, Pyoung Kim, Byung-Gee Kim] 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) fatty acids and hydroxy fatty acid, their biosynthetic pathways including thioesterase (Lreu_0335) from Lactobacillus reuteri DSM20016, β-hydroxyacyl-ACP dehydratase (fabZ) from Escherichia coli, and a P450 system (i.e., CYP153A from Marinobacter aquaeolei VT8 and camA/camB from Pseudomonas putida ATCC17453) were overexpressed. Acyl-CoA synthase (fadD) involved in fatty acid degradation by β-oxidation was also deleted in E. coli BW25113. The engineered E. coli FFA4 strain without the P450 system could produce 503.0mg/l of palmitic (C16) and 508.4mg/l of stearic (C18) acids, of which the amounts are ca. 1.6- and 2.3-fold higher than those of the wild type. On the other hand, the E. coli HFA4 strain including the P450 system for ω-hydroxylation could produce 211.7mg/l of ω-hydroxy palmitic acid, which was 42.1 ± 0.1% of the generated palmitic acid, indicating that the hydroxylation reaction was the rate-determining step for the HFA production. For the maximum production of ω-hydroxy palmitic acid, NADH, i.e., an essential cofactor for P450 reaction, was overproduced by the integration of NAD+-dependent formate dehydrogenase (FDH) from Candida boidinii into E. coli chromosome and the deletion of alcohol dehydrogenase (ADH). Finally, the NADH-level-optimized E. coli strain produced 610mg/l of ω-hydroxy palmitic acid (ω-HPA), which was almost a threefold increase in its yield compared to the same strain without NADH overproduction. Springer-Verlag Berlin Heidelberg, 2015 Free fatty acid nationallicence Hydroxy fatty acid nationallicence Metabolic engineering nationallicence P450 monooxygenase nationallicence Cofactor optimization nationallicence Sung Changmin Interdisciplinary Program for Bioengineering, Seoul National University, Seoul, South Korea aut Jung Eunok School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut Choi Kwon-Young Department of Environmental Engineering, College of Engineering, Ajou University, Suwon, Gyeonggi-do, South Korea aut Bae Jin-hyung School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut Kim Minsuk School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut Kim Joonwon School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut Kim Eun-Jung School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea aut Kim Pyoung Bio Control Research Institute, JBF, 495 Ipmyeon-ro, Ipmyeon, Gokseong-gun, Jeonnam, South Korea aut Kim Byung-Gee Interdisciplinary Program for Bioengineering, Seoul National University, Seoul, South Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6667-6676 0175-7598 99:16<6667 2015 99 253 https://doi.org/10.1007/s00253-015-6630-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6630-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sung Changmin Interdisciplinary Program for Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Jung Eunok School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Choi Kwon-Young Department of Environmental Engineering, College of Engineering, Ajou University, Suwon, Gyeonggi-do, South Korea aut NATIONALLICENCE 700 1- Bae Jin-hyung School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Minsuk School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Joonwon School of Chemical and Biological Engineering, Institute of Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Eun-Jung School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 700 1- Kim Pyoung Bio Control Research Institute, JBF, 495 Ipmyeon-ro, Ipmyeon, Gokseong-gun, Jeonnam, South Korea aut NATIONALLICENCE 700 1- Kim Byung-Gee Interdisciplinary Program for Bioengineering, Seoul National University, Seoul, South Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6667-6676 0175-7598 99:16<6667 2015 99 253