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

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Abstract

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.

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Data availability

All datasets obtained for this study are included in the manuscript/supplementary material.

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Funding

This research was supported by the 2020 Team Innovation Project from the Fundamental Scientific Research Special Capital Fund of the National Universities, China (100034/1301030160) and the Natural Science Foundation of China (NSFC) Project (grant number 32172145). Fundamental Research Funds for Central Universities of the Central South University,100034/1301030160,Yong Hong Meng,Natural Science Foundation of China (NSFC) Project,32172145,Yong Hong Meng

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Authors and Affiliations

  1. The Engineering Research Center for High-Valued Utilization of Fruit Resources in Western China, Ministry of EducationNational Research & Development Center of Apple Processing TechnologyCollege of Food Engineering and Nutritional Science, Shaanxi Normal University, 620 West Changan Avenue, Changan, Xian, 710119, People’s Republic of China

    Qi Hang Chen, Ching Yuan Hu & Yong Hong Meng

  2. Xian Healthful Biotechnology Co., Ltd, HangTuo Road, Changan, Xian, 710100, People’s Republic of China

    Dao Tao Xie & Shan Qiang

  3. Department of Human Nutrition, Food and Animal Sciences, College of Tropical Agriculture and Human Resources, University of Hawaii at Manoa, 1955 East-West Road, AgSci. 415J, Honolulu, HI, 96822, USA

    Ching Yuan Hu

Authors
  1. Qi Hang Chen
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  2. Dao Tao Xie
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  3. Shan Qiang
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  4. Ching Yuan Hu
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  5. Yong Hong Meng
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Contributions

QC, DX, SQ, and YM designed the experiments. QC performed the experiments. QC and YM were responsible for data processing and analysis. QC, YM, and CH wrote and revised manuscripts.

Corresponding author

Correspondence to Yong Hong Meng.

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Chen, Q.H., Xie, D.T., Qiang, S. et al. Developing efficient vanillin biosynthesis system by regulating feruloyl-CoA synthetase and enoyl-CoA hydratase enzymes. Appl Microbiol Biotechnol 106, 247–259 (2022). https://doi.org/10.1007/s00253-021-11709-w

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  • DOI: https://doi.org/10.1007/s00253-021-11709-w

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