Biosynthesis of C12 Fatty Alcohols by Whole Cell Biotransformation of C12 Derivatives Using Escherichia coli Two-cell Systems Expressing CAR and ADH

• Published in: Biotechnology and bioprocess engineering Vol. 26; no. 3; pp. 392 - 401
• Main Authors: Cha, Tae-Yong, Yong, Yuk, Park, HyunA, Yun, Hye-Jung, Jeon, Wooyoung, Ahn, Jung-Oh, Choi, Kwon-Young
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01-06-2021; Springer Nature B.V; 한국생물공학회
• Subjects: Acids; Alcohols; Biosynthesis; Biotechnology; Biotransformation; Chemistry; Chemistry and Materials Science; Conversion; DnaK protein; E coli; Enzymes; Escherichia coli; Hydrogen bonding; Hydroxyl groups; Industrial and Production Engineering; Lauric acid; Research Paper; Substrates; 생물공학; whole cell biotransformation; carboxylic acid reductase; alcohol dehydrogenase; reductive metabolites; two cell reactions
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-020-0239-7

In this study, the conversions of 1-dodecanoic, ω-hydroxydodecanoic acid and α,ω-dodecanedioic acid using whole cell biotransformation of
Escherichia coli
BW25113
Δ
fadD
expressing CAR and ADH enzymes were demonstrated. First 13 CAR enzymes were examined for 1-dodecanoic acid reduction, and CAR encoded by
mab4714
from
Mycobacterium abscessus
showed the highest conversion of 53.1% in single cells of heterologous CAR and endogenous ADH. For a better conversion, the host cells were engineered to simultaneously express
Yarrowia lipolytica
ADH2 with the GroES/EL-DnaK/J/E chaperone in a single host system. In addition, two-cell system using two strains of
E. coli
expressing CAR-Sfp and ADH-GroES/EL-DnaK/J/E was also investigated. In results, additional ADH expression was not effective in a single host system, whereas two cell system significantly increased α,ω-dodecanedioic acid conversion by total 71.3%; α,ω-dodecanediol (68.2%) and ω-hydroxydodecanoic acid (3.1%), respectively. Interestingly, the MAB4714 CAR enzyme could converted ω-hydroxydodecanoic acid into α,ω-dodecanediol up to 97.2% conversion in 17 h (12.4 mg/L/h). Finally, structural understanding of the higher activity against ω-hydroxydodecanoic was understood by docking simulations which suggested hydrogen-bonding interactions between ω-hydroxyl group and polar residues such as Gln434 and Thr285 were holding the substrate tightly with more stable positioning in the active site.