Multi-level engineering of Baeyer-Villiger monooxygenase-based Escherichia coli biocatalysts for the production of C9 chemicals from oleic acid
- 주제(키워드) Baeyer-Villiger monooxygenase , Plasmid copy number , Gene expression control , Escherichia coli , Biocatalysis , Fatty acids
- 주제(기타) Biotechnology & Applied Microbiology
- 설명문(일반) [Seo, Eun-Ji; Woo, Ji-Min; Park, Jin-Byung] Ewha Womans Univ, Dept Food Sci & Engn, Seoul 03760, South Korea; [Kang, Chae Won; Jang, Sungho; Jung, Gyoo Yeol] Pohang Univ Sci & Technol, Dept Chem Engn, 77 Cheongam Ro, Pohang 37673, South Korea; [Yeon, Young Joo] Gangneung Wonju Natl Univ, Dept Biochem Engn, Kangnung 25457, South Korea; [Jung, Gyoo Yeol] Pohang Univ Sci & Technol, Sch Interdisciplinary Biosci & Bioengn, Pohang 37673, South Korea
- 등재 SCIE, SCOPUS
- 발행기관 ACADEMIC PRESS INC ELSEVIER SCIENCE
- 발행년도 2019
- URI http://www.dcollection.net/handler/ewha/000000160063
- 본문언어 영어
- Published As http://dx.doi.org/10.1016/j.ymben.2019.03.012
- PubMed https://pubmed.ncbi.nlm.nih.gov/30953778
초록/요약
Whole-cell biotransformation is one of the promising alternative approaches to microbial fermentation for producing high-value chemicals. Baeyer-Villiger monooxygenase (BVMO)-based Escherichia coil biocatalysts have been engineered to produce industrially relevant C9 chemicals, such as n-nonanoic acid and 9-hydroxynonanoic acid, from a renewable long-chain fatty acid. The key enzyme in the biotransformation pathway (i.e., BVMO from Pseudomonans putida KT2440) was first engineered, using structure modeling-based design, to improve oxidative and thermal stabilities. Using a stable and tunable plasmid (STAPL) system, E. coli. host cells were engineered to have increased plasmid stability and homogeneity of the recombinant E. coli. population, as well as to optimize the level of BVMO expression. Multi-level engineering of the key enzyme in host cells, allowed recombinant E. coli. expressing a fatty acid double-bond hydratase, a long-chain secondary alcohol dehydrogenase, and the engineered BVMO from P. putida KT2440 (i.e., E6BVMO_C302L/M340L), to ultimately produce C9 chemicals (i.e., n-nonanoic acid and 9-hydroxynonanoic acid) from oleic acid, with a yield of up to 6 mmoL/g dry cells. This yield was 2.4-fold greater than the yield in the control strain before engineering. Therefore, this study will contribute to the development of improved processes for the biosynthesis of industrially relevant medium chain fatty acids via whole-cell biocatalysis.
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