Improving catalytic activity of the Baeyer-Villiger monooxygenase-based Escherichia coli biocatalysts for the overproduction of (Z)-11-(heptanoyloxy) undec-9-enoic acid from ricinoleic acid
- 주제(기타) Multidisciplinary Sciences
- 설명문(일반) [Woo, Ji-Min; Jeon, Eun-Yeong; Seo, Eun-Ji; Park, Jin-Byung] Ewha Womans Univ, Dept Food Sci & Engn, Seoul 03760, South Korea; [Seo, Joo-Hyun] Sun Moon Univ, Dept BT Convergent Pharmaceut Engn, Asan 31460, South Korea; [Lee, Dong-Yup] Sungkyunkwan Univ, Sch Chem Engn, Suwon 16419, South Korea; [Yeon, Young Joo] Gangneung Wonju Natl Univ, Dept Biochem Engn, Kangnung 25457, South Korea; [Park, Jin-Byung] Ewha Womans Univ, Inst Mol Microbiol & Biosyst Engn, Seoul 03760, South Korea
- 등재 SCIE, SCOPUS
- 발행기관 NATURE PUBLISHING GROUP
- 발행년도 2018
- URI http://www.dcollection.net/handler/ewha/000000151620
- 본문언어 영어
- Published As http://dx.doi.org/10.1038/s41598-018-28575-8
초록/요약
Baeyer-Villiger monooxygenases (BVMOs) can be used for the biosynthesis of lactones and esters from ketones. However, the BVMO-based biocatalysts are not so stable under process conditions. Thereby, this study focused on enhancing stability of the BVMO-based biocatalysts. The biotransformation of ricinoleic acid into (Z)-11-(heptanoyloxy) undec-9-enoic acid by the recombinant Escherichia coli expressing the BVMO from Pseudomonas putida and an alcohol dehydrogenase from Micrococcus luteus was used as a model system. After thorough investigation of the key factors to influence stability of the BVMO, Cys302 was identified as an engineering target. The substitution of Cys302 to Leu enabled the engineered enzyme (i.e., E6BVMO(C302L)) to become more stable toward oxidative and thermal stresses. The catalytic activity of E6BVMO(C302L)-based E. coli biocatalysts was also greater than the E6BVMO-based biocatalysts. Another factor to influence biocatalytic performance of the BVMO-based whole-cell biocatalysts was availability of carbon and energy source during biotransformations. Glucose feeding into the reaction medium led to a marked increase of final product concentrations. Overall, the bioprocess engineering to improve metabolic stability of host cells in addition to the BVMO engineering allowed us to produce (Z)-11-(heptanoyloxy) undec-9-enoic acid to a concentration of 132 mM (41 g/L) from 150 mM ricinoleic acid within 8 h.
more