Metabolic engineering ofE. colifor producing phloroglucinol from acetate

文献类型： 外文期刊

第一作者： Yu, Shengzhu

作者： Yu, Shengzhu;Guo, Liwei;Zhao, Luyao;Chen, Zhenya;Huo, Yixin;Huo, Yixin

作者机构：

关键词： Phloroglucinol; PhlD; Acetate; CRISPRi; Optimization; Low-cost production

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2020 年 104 卷 18 期

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收录情况： SCI

摘要： Phloroglucinol is a three-hydroxyl phenolic compound and has diverse physiological and pharmacological activities such as antivirus and anti-inflammatory activities. Chemical synthesis of phloroglucinol suffered from many drawbacks such as high cost and environmental pollution. To avoid the above issues, microbial phloroglucinol biosynthesis was successfully accomplished in this study, while the abundant and low-cost acetate was used as the main carbon source. Firstly, the toxicity of phloroglucinol was tested, andE. coliBL21(DE3) could tolerate 5 g/L phloroglucinol. The ability of phloroglucinol synthase (PhlD) for catalyzing malonyl-CoA to phloroglucinol was confirmed, andE. coliBL21(DE3) expressing PhlD and acetyl-CoA carboxylase (ACCase) could produce 1107 +/- 12 mg/L phloroglucinol from glucose. Then,E. coliBL21(DE3) was engineered to utilize acetate to produce 228 +/- 15 mg/L phloroglucinol. Then, the endogenous citrate synthase (GltA) which could catalyze oxaloacetate and acetyl-CoA generated from acetate to citrate was knocked down by CRISPRi system in order to enhance the carbon flux for phloroglucinol production, and the titer was improved to 284 +/- 8 mg/L. This work demonstrated that acetate could be used as low-cost substrate to achieve the biosynthesis of phloroglucinol and provided an example of effective utilization of acetate.

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