Developing efficient vanillin biosynthesis system by regulating feruloyl-CoA synthetase and enoyl-CoA hydratase enzymes

• Published in: Applied microbiology and biotechnology Vol. 106; no. 1; pp. 247 - 259
• Main Authors: Chen, Qi Hang, Xie, Dao Tao, Qiang, Shan, Hu, Ching Yuan, Meng, Yong Hong
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 2022; Springer; Springer Nature B.V
• Subjects: Benzaldehydes; Biomedical and Life Sciences; Biosynthesis; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Coenzyme A Ligases - genetics; E coli; Enoyl-CoA hydratase; Enoyl-CoA Hydratase - genetics; Enzymes; Escherichia; Escherichia coli; Escherichia coli - genetics; Ferulic acid; Kinases; Life Sciences; Methods; Microbial Genetics and Genomics; Microbiology; Physiological aspects; Production processes; Streptomyces; Vanillin; China; Enoyl-CoA hydratase; Whole-cell catalytic system; Vanillin; Feruloyl-CoA synthetase; Cell-free catalytic system; Ferulic acid
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-021-11709-w

Vanillin is one of the most commonly used natural-occurring flavors in the world. This study successfully constructed an efficient whole-cell catalytic system for vanillin biosynthesis from ferulic acid by regulating feruloyl-CoA synthetase (FCS) and enoyl-CoA hydratase (ECH). First, we constructed an efficient cell-free catalytic system with FCS-Str ( fcs from Streptomyces sp. V-1) and ECH-Str ( ech from Streptomyces sp. V-1) combination at 1:1. The efficient cell-free catalytic system provided necessary strategies for optimizing the whole-cell catalytic system. Then, we constructed the recombinant Escherichia coli by heterologously expressing the fcs -Str and ech -Str combination. Moreover, E. coli JM109 was a better recombinant Escherichia coli than E. coli BL21 with higher vanillin production. Finally, we first adjusted the ratio of FCS and ECH in E. coli JM109 to 1:1 using two copies of fcs -Str. For higher vanillin production, we further optimized the induction conditions of E. coli JM109 to increase the amount of FCS and ECH. The optimized E. coli JM109-FE-F constructed in this study has the highest vanillin synthesis rate of converting 20 mM ferulic acid to 15 mM vanillin in 6 h among all of the E. coli catalytic systems. Our study made a significant contribution to the construction of the vanillin biosynthesis system and provided a valuable strategy for increasing vanillin production. Key points • The efficient cell-free vanillin biosynthesis system was constructed by FCS-Str and ECH-Str combination at 1:1. • Escherichia coli JM109 was determined as a better recombinant Escherichia coli than E. coli BL21 with higher vanillin production. • Escherichia coli JM109-FE-F with two copies of fcs-Str and one copy of ech-Str has the highest catalytic efficiency for vanillin production.