微生物合成植物天然产物的细胞工厂设计与构建

doi:10.16085/j.issn.1000-6613.2020-2101

摘要/Abstract

摘要：

植物天然产物结构多样，具有丰富的生理活性与功能。利用微生物细胞工厂生产来源稀缺、获取难度大的植物天然产物具有经济可行、环境友好等优势。本文系统介绍了萜烯、黄酮以及生物碱的生物合成途径及其关键酶，阐述了差异转录组学、同功酶挖掘等途径解析与重构的方法。指出关键酶改造、途径动态调控、代谢区室化与代谢网络再平衡是增大外源途径代谢通量、抑制副产物合成、降低产物毒性与菌株代谢负担、提高目标产物合成能力的有效策略。提出了解析合成植物天然产物关键酶在微生物中的催化特异性机制、开发外源途径的高效组装方法等进一步提高微生物细胞工厂生产效率的建议。

关键词: 植物天然产物, 微生物细胞工厂, 途经重构, 合成生物学

Abstract:

Plant natural products possess diverse structures, physiological activities and functions. The use of microbial cell factories to produce plant natural products with scarce sources and difficult to obtain has the advantages of economic viability and environmental friendliness. This paper systematically introduces the biosynthetic pathways and key enzymes of terpenes, flavonoids and alkaloids, and expounds the decipher and reconstruction methods of their biosynthesis pathways including transcriptomics and isozyme mining. It is pointed out that enzyme engineering, dynamic control, metabolic compartmentation and metabolic network rebalance are effective strategies to increase the metabolic flux of exogenous pathways, inhibit by-product synthesis, reduce product toxicity and strain metabolic burden, and improve the synthesis ability of target products. Suggestions are put forward to further improve the production efficiency of microbial cell factories, such as analyzing the catalytic specific mechanism of plant enzymes in microorganisms, and developing efficient assembly methods for exogenous pathways.

Key words: plant natural products, microbial cell factories, pathway reconstruction, synthetic biology

中图分类号:

Q819

孙文涛, 李春. 微生物合成植物天然产物的细胞工厂设计与构建[J]. 化工进展, 2021, 40(3): 1202-1214.

SUN Wentao, LI Chun. Design and construction of microbial cell factory for biosynthesis of plant natural products[J]. Chemical Industry and Engineering Progress, 2021, 40(3): 1202-1214.

图/表 6

图1 萜烯化合物合成途径Acetyl-CoA—乙酰辅酶A；MVA—甲羟戊酸；MEP—2-甲基-D-赤藓醇-4-磷酸；IPP—异戊二烯焦磷酸；DMAPP—焦磷酸二甲基烯丙酯；GPP—牻牛儿基焦磷酸；GGPP—牻牛儿基牻牛儿基焦磷酸；FPP—法尼烯焦磷酸；OA—齐墩果酸；PPD—原人参二醇

图2 植物黄酮类化合物合成途径TAL—酪氨酸胺裂解酶；4CL—4-香豆酰辅酶A连接酶；CHS—查耳酮合酶；CHI—查耳酮异构酶；P450—CYP450氧化酶；OMT—氧甲基转移酶；UGT—UDP-葡萄糖醛酸基转移酶

图3 苄基异喹啉生物碱合成途径TYDC—酪氨酸脱羧酶；TyrAT—酪氨酸转氨酶；DODC—多巴胺脱羧酶；3OHase—色胺羟化酶；4-HPPDC—4-羟基苯丙酮酸脱羧酶；NCS—去甲乌药碱合酶；6OMT—去甲乌药碱6-O-甲基转移酶；CNMT—乌药碱-N-甲基转移酶；4OMT—3-羟基-N-甲基-乌药碱-4-O-甲基转移酶

图4 单萜吲哚生物碱合成途径DMAPP—焦磷酸二甲基烯丙酯；IPP—异戊二烯焦磷酸；SLS—马钱子苷合酶；STR—异胡豆苷合酶

图5 植物天然产物合成途径的重构策略

图6 植物天然产物合成途径的强化策略

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