大肠杆菌乙酰辅酶A代谢调控及其应用研究进展

doi:10.16085/j.issn.1000-6613.2019-0081

摘要/Abstract

摘要：

利用生物法合成生物基化学品具有高效、绿色、可持续发展等优势。乙酰辅酶A作为细胞内物质代谢的重要中间产物，是利用生物转化法合成许多生物基化学品的重要前体，在微生物碳代谢过程中发挥着枢纽作用。本文综述了大肠杆菌乙酰辅酶A的合成、代谢调控策略及其重要应用，重点总结了乙酰辅酶A的合成途径及近期发展的提高乙酰辅酶A胞内通量的代谢调控策略，包括乙酸途径的代谢调控、丙酮酸合成乙酰辅酶A途径的代谢调控、中心碳代谢途径的代谢调控、β氧化合成乙酰辅酶A途径的代谢调控和乙酰辅酶A合成新途径的发掘，进一步展望了提高乙酰辅酶A供给的策略，利用基因组编辑技术构建合成乙酰辅酶A为前体化学品细胞工厂的方法。

关键词: 乙酰辅酶A, 大肠杆菌, 代谢工程, 通量

Abstract:

Biosynthesis of chemicals has the advantages of high efficiency, green and sustainable development. Acetyl coenzyme A, as an important intermediate of cellular metabolism in cells, is an important precursor for the synthesis of many biochemicals, and has played a vital role in the process of microbial carbon metabolism. Here we reviewed the synthesis and strategies of metabolic regulation of acetyl coenzyme A in Escherichia coli and its important applications. We also summarized the synthesis pathway of acetyl coenzyme A and the recent development of metabolic regulation strategies to increase the intracellular metabolic flux of acetyl coenzyme A production, these including metabolic regulation of the acetyl coenzyme A synthesized by acetic acid and pyruvate, metabolic regulation of the central carbon metabolic pathway and acetyl coenzyme A synthesized by beta oxidation pathway, and discovery of new pathways for acetyl coenzyme A synthesis. Finally, we prospected the feasible strategies of increasing acetyl coenzyme A supply and discussed methods of constructing cell factories for synthesizing acetyl coenzyme A as precursor chemicals by genome editing technology.

Key words: acetyl coenzyme A, Escherichia coli, metabolic engineering, flux

中图分类号:

Q756

陈露,刘丁玉,汪保卫,赵玉姣,贾广韬,陈涛,王智文. 大肠杆菌乙酰辅酶A代谢调控及其应用研究进展[J]. 化工进展, 2019, 38(9): 4218-4226.

Lu CHEN,Dingyu LIU,Baowei WANG,Yu jiao ZHAO,Guangtao JIA,Tao CHEN,Zhiwen WANG. Advances in acetyl coenzyme A metabolic engineering with Escherichia coli[J]. Chemical Industry and Engineering Progress, 2019, 38(9): 4218-4226.

图/表 4

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途径	基因操作	产物	产量		参考文献
途径	基因操作	产物	对照	工程菌株	参考文献
乙酸途径	过表达hyc和acs	聚-β-羟丁酸	0.55mg聚-β-羟丁酸/mg 葡萄糖	5.34mg聚-β-羟丁酸/mg葡萄糖	[6]
	敲除ackA-pta和nuo	乙酰辅酶A	产量提高31%		[12]
	敲除ackA-pta，过表达acs	丙二酰辅酶A	产量提高两倍		[13]
丙酮酸途径	过表达aceEF-lpdA	琥珀酸	7.0mmol/L	17.5mmol/L	[18]
	过表达aceEF-lpdA	琥珀酸	4.2mmol/L	12.2mmol/L	[19]
	敲除iclR，fadR	琥珀酸	0.5mol/mol	0.98mol/mol	[20]
	敲除fnr	1-丁醇	130mg/L	373mg/L	[22]
	过表达aceEF-IpdA，panK	乙酸异戊酯	0.1mmol/L	0.18mmol/L	[25]
糖酵解途径	过表达tpiA，fbaA	聚-β-羟丁酸	0.4g/L	1.6g/L	[27]
三羧酸循环途径	敲除fumB/C 或sucC	丙二酰辅酶A	2.55μmol/gDW	4.43μmol/gDW	[28]
	敲除sdhA和citE	丙二酰辅酶A	0.95nmol/mg	2.60nmol/mg	[29]
乙醛酸途径	敲除iclR	3-羟基丙酸	产量提高1.9倍		[30]
	敲除icdA	香草醛	0.79g/L	2.06g/L	[31]
磷酸戊糖途径	过表达tktA	聚-β-羟丁酸	产量提高1.7倍		[32]
	过表达zwf	聚-β-羟丁酸	产量提高41%		[33]
2-酮-3-脱氧-6-磷酸葡糖酸途径	过表达edd、eda 和aceEF	聚-β-羟丁酸	产量提高81.1%		[35]
β氧化途径	敲除fadR	3-羟基丙酸	产量达到52g/L		[37]
非氧化酵解途径	过表达xpk（Bifidobacterium adolescentis）和fba	聚-β-羟丁酸	2.36g/L	4.69g/L	[40]
	过表达xpk（Bifidobacterium adolescentis）	丙酮	0.38mol/mol	0.47mol/mol	[41]

途径	基因操作	产物	产量		参考文献
途径	基因操作	产物	对照	工程菌株	参考文献
乙酸途径	过表达hyc和acs	聚-β-羟丁酸	0.55mg聚-β-羟丁酸/mg 葡萄糖	5.34mg聚-β-羟丁酸/mg葡萄糖	[6]
	敲除ackA-pta和nuo	乙酰辅酶A	产量提高31%		[12]
	敲除ackA-pta，过表达acs	丙二酰辅酶A	产量提高两倍		[13]
丙酮酸途径	过表达aceEF-lpdA	琥珀酸	7.0mmol/L	17.5mmol/L	[18]
	过表达aceEF-lpdA	琥珀酸	4.2mmol/L	12.2mmol/L	[19]
	敲除iclR，fadR	琥珀酸	0.5mol/mol	0.98mol/mol	[20]
	敲除fnr	1-丁醇	130mg/L	373mg/L	[22]
	过表达aceEF-IpdA，panK	乙酸异戊酯	0.1mmol/L	0.18mmol/L	[25]
糖酵解途径	过表达tpiA，fbaA	聚-β-羟丁酸	0.4g/L	1.6g/L	[27]
三羧酸循环途径	敲除fumB/C 或sucC	丙二酰辅酶A	2.55μmol/gDW	4.43μmol/gDW	[28]
	敲除sdhA和citE	丙二酰辅酶A	0.95nmol/mg	2.60nmol/mg	[29]
乙醛酸途径	敲除iclR	3-羟基丙酸	产量提高1.9倍		[30]
	敲除icdA	香草醛	0.79g/L	2.06g/L	[31]
磷酸戊糖途径	过表达tktA	聚-β-羟丁酸	产量提高1.7倍		[32]
	过表达zwf	聚-β-羟丁酸	产量提高41%		[33]
2-酮-3-脱氧-6-磷酸葡糖酸途径	过表达edd、eda 和aceEF	聚-β-羟丁酸	产量提高81.1%		[35]
β氧化途径	敲除fadR	3-羟基丙酸	产量达到52g/L		[37]
非氧化酵解途径	过表达xpk（Bifidobacterium adolescentis）和fba	聚-β-羟丁酸	2.36g/L	4.69g/L	[40]
	过表达xpk（Bifidobacterium adolescentis）	丙酮	0.38mol/mol	0.47mol/mol	[41]

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