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

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Abstract

In this study, the conversions of 1-dodecanoic, ω-hydroxydodecanoic acid and α,ω-dodecanedioic acid using whole cell biotransformation of Escherichia coli BW25113ΔfadD 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.

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Acknowledgements

This work was supported by the Industrial Strategic Technology Development program (No. 20002734).

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  1. These two authors contributed equally

Authors and Affiliations

  1. Department of Environmental Engineering, College of Engineering, Ajou University, Suwon, 16499, Korea

    Tae-Yong Cha, HyunA Park, Hye-Jung Yun & Kwon-Young Choi

  2. Department of Environmental and Safety Engineering, College of Engineering, Ajou University, Suwon, 16499, Korea

    Yuk Yong & Kwon-Young Choi

  3. Biotechnology Process Engineering Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Cheongju, 28116, Korea

    Wooyoung Jeon & Jung-Oh Ahn

  4. Department of Bioprocess Engineering, University of Science and Technology (UST) of Korea, Daejeon, 34113, Korea

    Jung-Oh Ahn

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Cha, TY., Yong, Y., Park, H. et al. Biosynthesis of C12 Fatty Alcohols by Whole Cell Biotransformation of C12 Derivatives Using Escherichia coli Two-cell Systems Expressing CAR and ADH. Biotechnol Bioproc E 26, 392–401 (2021). https://doi.org/10.1007/s12257-020-0239-7

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