Highly efficient biosynthesis of beta-caryophyllene with a new sesquiterpene synthase from tobacco

文献类型： 外文期刊

第一作者： Cheng, Tao

作者： Cheng, Tao;Zhang, Kai;Guo, Jing;Xian, Mo;Zhang, Rubing;Yang, Qing;Li, Yiting;Xian, Mo;Zhang, Rubing

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关键词： beta-Caryophyllene; Escherichia coli; Mevalonate pathway; Fed-batch fermentation; In situ extraction

期刊名称：BIOTECHNOLOGY FOR BIOFUELS AND BIOPRODUCTS

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年卷期： 2022 年 15 卷 1 期

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收录情况： SCI

摘要： Background: beta-Caryophyllene, a kind of bicyclic sesquiterpene, is mainly used as a spice in the food and cosmetic industries. Furthermore, it also has significant value in the pharmaceutical industry and is now considered to be used as a new fuel. As a chemical energy heterotrophic microorganism, Escherichia coli can produce a large amount of acetyl-CoA through aerobic respiration, and acetyl-CoA is the common precursor substance in the biosynthesis of all terpenoids. Therefore, E. coli has the potential to be a cell factory to produce terpenoids. Results: A new gene of beta-caryophyllene synthase (TPS7) was found by analyzing the genome of Nicotiana tabacum L. using bioinformatics methods. The gene was overexpressed in engineered E. coli with a heterogeneous mevalonate (MVA) pathway to build a recombinant strain CAR1. Subsequent cultivation experiments in shake flask of engineered strain CAR1 verified that 16.1 mg/L beta-caryophyllene was detected from the fermentation broth in the shake flask after induction for 24 h with IPTG. The toxic by-product of farnesyl acetate was detected during the process, and CAR1 showed a heavily cellular accumulation of product. We constructed an engineered strain CAR2, in which the downstream genes of the MVA pathway were integrated into the E. coli chromosome, successfully increasing P-caryophyllene production to 100.3 mg/L. The highest production of beta-caryophyllene during the fed-batch fermentation was 4319 mg/L. Then we employed in situ extraction fermentation to successfully increase the production of beta-caryophyllene by 20% to 5142 mg/L. Conclusion: A new sesquiterpene synthase,TPS7, from tobacco was found to be able to produce beta-caryophyllene with high efficiency. Based on this, an engineered E. coli was constructed to produce a much higher concentration of beta-caryophyllene than the previous studies. During the fermentation process, we observed that beta-caryophyllene tends to accumulate in intracellular space, which will eventually influence the activity of engineered E. coli. As a result, we solved this by metabolism regulation and in situ extractive fermentation.

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