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A novel process for obtaining pinosylvin using combinatorial bioengineering in Escherichia coli

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Abstract

Pinosylvin as a bioactive stilbene is of great interest for food supplements and pharmaceuticals development. In comparison to conventional extraction of pinosylvin from plant sources, biosynthesis engineering of microbial cell factories is a sustainable and flexible alternative method. Current synthetic strategies often require expensive phenylpropanoic precursor and inducer, which are not available for large-scale fermentation process. In this study, three bioengineering strategies were described to the development of a simple and economical process for pinosylvin biosynthesis in Escherichia coli. Firstly, we evaluated different construct environments to give a highly efficient constitutive system for enzymes of pinosylvin pathway expression: 4-coumarate: coenzyme A ligase (4CL) and stilbene synthase (STS). Secondly, malonyl coenzyme A (malonyl-CoA) is a key precursor of pinosylvin bioproduction and at low level in E. coli cell. Thus clustered regularly interspaced short palindromic repeats interference (CRISPRi) was explored to inactivate malonyl-CoA consumption pathway to increase its availability. The resulting pinosylvin content in engineered E. coli was obtained a 1.9-fold increase depending on the repression of fabD (encoding malonyl-CoA-ACP transacylase) gene. Eventually, a phenylalanine over-producing E. coli consisting phenylalanine ammonia lyase was introduced to produce the precursor of pinosylvin, trans-cinnamic acid, the crude extraction of cultural medium was used as supplementation for pinosylvin bioproduction. Using these combinatorial processes, 47.49 mg/L pinosylvin was produced from glycerol.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant Nos. 31071837, 31272217, and 31372116); the Projects of Science and Technology of Guangdong Province (Grant No. 2013B010404041 and 2014B050505018).

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Authors and Affiliations

  1. Department of Bioengineering, College of Food Science and Institute of Food Biotechnology, South China Agricultural University, 483 Wushan Road, Tianhe District, Guangzhou, 510640, China

    Jing-long Liang, Li-qiong Guo, Jun-fang Lin, Ze-qi He & Fa-ji Cai

  2. Joint Research and Development Center for Natural Products of Alchemy Biotechnology Co. Ltd. and South China Agricultural University, Guangzhou, 510640, China

    Jing-long Liang, Li-qiong Guo & Jun-fang Lin

  3. The School of Chemistry and Pharmaceutical Engineering, Qi-Lu University of Technology, Jinan, 250353, China

    Jun-fei Chen

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  1. Jing-long Liang
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  2. Li-qiong Guo
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  3. Jun-fang Lin
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  4. Ze-qi He
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Correspondence to Jun-fang Lin.

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Liang, Jl., Guo, Lq., Lin, Jf. et al. A novel process for obtaining pinosylvin using combinatorial bioengineering in Escherichia coli . World J Microbiol Biotechnol 32, 102 (2016). https://doi.org/10.1007/s11274-016-2062-z

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  • DOI: https://doi.org/10.1007/s11274-016-2062-z

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