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[通过工程改造酿酒酵母动态控制ERG20表达以提高单萜产量]

[Dynamic control of ERG20 expression to improve production of monoterpenes by engineering Saccharomyces cerevisiae].

作者信息

Li Rong-Sheng, Wang Dong, Shi Yu-Song, Xu Li-Ping, Zhang Xue-Li, Wang Kou, Dai Zhu-Bo

机构信息

School of Pharmacy and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University Kunming 650500, China Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences Tianjin 300308,China Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China.

Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences Tianjin 300308,China Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2022 Feb;47(4):897-905. doi: 10.19540/j.cnki.cjcmm.20210913.101.

Abstract

Monoterpenes are widely used in cosmetics, food, medicine, agriculture and other fields. With the development of synthetic biology, it is considered as a potential way to create microbial cell factories to produce monoterpenes. Engineering Saccharomyces cerevisiae to produce monoterpenes has been a research hotspot in synthetic biology. In S. cerevisiae, the production of geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) is catalyzed by a bifunctional enzyme farnesyl pyrophosphate synthetase(encoded by ERG20 gene) which is inclined to synthesize FPP essential for yeast growth. Therefore, reasonable control of FPP synthesis is the basis for efficient monoterpene synthesis in yeast cell factories. In order to achieve dynamic control from GPP to FPP biosynthesis in S. cerevisiae, we obtained a novel chassis strain HP001-pERG1-ERG20 by replacing the ERG20 promoter of the chassis strain HP001 with the promoter of cyclosqualene cyclase(ERG1) gene. Further, we reconstructed the metabolic pathway by using GPP and neryl diphosphate(NPP), cis-GPP as substrates in HP001-pERG1-ERG20. The yield of GPP-derived linalool increased by 42.5% to 7.6 mg·L(-1), and that of NPP-derived nerol increased by 1 436.4% to 8.3 mg·L(-1). This study provides a basis for the production of monoterpenes by microbial fermentation.

摘要

单萜广泛应用于化妆品、食品、医药、农业等领域。随着合成生物学的发展,构建微生物细胞工厂来生产单萜被认为是一种具有潜力的方法。工程改造酿酒酵母以生产单萜一直是合成生物学领域的研究热点。在酿酒酵母中,香叶基焦磷酸(GPP)和法尼基焦磷酸(FPP)的合成由一种双功能酶法尼基焦磷酸合成酶(由ERG20基因编码)催化,该酶倾向于合成对酵母生长至关重要的FPP。因此,合理控制FPP的合成是酵母细胞工厂高效合成单萜的基础。为了实现酿酒酵母中从GPP到FPP生物合成的动态控制,我们通过用环鲨烯环化酶(ERG1)基因的启动子替换底盘菌株HP001的ERG20启动子,获得了一种新型底盘菌株HP001-pERG1-ERG20。此外,我们在HP001-pERG1-ERG20中以GPP、橙花基二磷酸(NPP)、顺式-GPP为底物重建了代谢途径。GPP衍生的芳樟醇产量提高了42.5%,达到7.6 mg·L⁻¹,NPP衍生的橙花醇产量提高了1436.4%,达到8.3 mg·L⁻¹。本研究为微生物发酵生产单萜提供了依据。

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