Construction and application of microbial cell factories for production of bulk chemicals

doi:10.12211/2096-8280.2020-049

Abstract

Abstract:

With the development of synthetic biology, more and more bulk chemicals can be produced through microbial cell factories, which can avoid the dependence on petroleum resources and decrease energy cost and pollution. In this review, the key technologies for construction of microbial cell factories were firstly introduced, including genome editing, simultaneous modulation of multiple genes, protein scaffold, gene dynamic modulation and high throughput screening technologies. How to characterize the metabolic regulation mechanisms for efficient production of chemicals was then introduced in three aspects: carbon metabolism, energy metabolism and physiological metabolism, and succinate cell factory was used as an example. Successful bulk chemical cell factories in recent years were then summarized, including L-alanine, L-methionine, succinate, D-lactate, malonate, L-malate, glutarate, adipate, 1,3-propanediol, 1,4-butanediol, isobutanol, etc. In the future, further increasing the efficiency of substrates utilization and broadening the range of products will be the directions of development of microbial cell factories, but the design and engineering of new enzymes are the key bottleneck limiting the design of metabolic pathways. It is believed that with the deepening of research, microbial cell factories will be more widely used in the production of bulk chemicals besides chemical methods.

Key words: synthetic biology, bulk chemicals, cell factories, metabolic engineering, metabolic regulation, genome editing

摘要：

随着合成生物学技术的发展，越来越多的大宗化学品可通过微生物细胞工厂发酵生产，为摆脱石油资源依赖、节能减排提供了可能。本文首先介绍了细胞工厂构建所需的关键技术，包括基因组编辑技术、多基因同时调控技术、蛋白骨架技术、基因动态调控技术、高通量筛选技术。随后结合丁二酸细胞工厂这一具体案例，从物质代谢、能量代谢及细胞生理代谢三方面阐述了如何解析微生物高效合成化学品的代谢调控机制，为高效细胞工厂创建奠定理论基础。并对近年来成功构建的大宗化学品细胞工厂作了举例介绍，包括L-丙氨酸、L-甲硫氨酸、丁二酸、D-乳酸、丙二酸、L-苹果酸、戊二酸、己二酸、1，3-丙二醇、1，4-丁二醇和异丁醇等。未来，进一步增加原料利用效率和拓宽产物范围是微生物细胞工厂的发展方向，但新酶设计与改造是限制代谢途径设计的主要瓶颈。相信随着研究的深入，微生物细胞工厂将在替代化学法生产大宗化学品方面有更广泛的应用。

关键词: 合成生物学, 大宗化学品, 细胞工厂, 代谢工程, 代谢调控, 基因组编辑

CLC Number:

YU Yong, ZHU Xinna, ZHANG Xueli. Construction and application of microbial cell factories for production of bulk chemicals[J]. Synthetic Biology Journal, 2020, 1(6): 674-684.

于勇, 朱欣娜, 张学礼. 大宗化学品细胞工厂的构建与应用[J]. 合成生物学, 2020, 1(6): 674-684.

Figures/Tables 5

Fig. 1 Mechanisms of metabolic regulation for efficient production of chemicals

Fig. 2 Four rate-limiting steps for synthesis of succinate

Fig. 3 Regulation of glucose energy metabolism to solve reducing equivalent problem for synthesis of succinate

Fig. 4 Regulation of glycerol energy metabolism to solve energy problem for synthesis of succinate

Fig. 5 New osmotolerance mechanism of Escherichia coli(a) Under normal conditions (5% glucose), copper exists mostly in the form of less-toxic Cu(Ⅱ), and the CopA and Cus system was not induced; (b) Under high osmotic pressure (12% glucose), increasing free Cu(Ⅰ) in the periplasmic space activates the expression of copA, cusS, cusR and cusCFAB to expel toxic Cu(Ⅰ); (c) Under high osmotic pressure (12% glucose), expression levels of cusS and cusCFAB are increased to expel more Cu(Ⅰ) from the periplasmic space; (d) Osmotic protective substances such as methionine and cysteine can chelate valence copper ions.

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