大肠杆菌生产饲用氨基酸的研究进展

doi:10.12211/2096-8280.2021-042

摘要/Abstract

摘要：

随着畜牧业的快速发展，人们对畜牧饲料蛋白的需求日益剧增。由于人们对食品安全意识的增强，迫切需要开发安全、高效、可持续动物饲料蛋白的供应途径。由于氨基酸是组成蛋白质的基本单元，所以在饲料中添加氨基酸可以替代饲料中的蛋白质，为动物细胞生长发育提供足够的营养。因此，饲用氨基酸作为动物饲料食品添加剂被广泛应用，具有广阔的市场应用前景。利用合成生物学技术，工程化改造大肠杆菌，构建的细胞工厂，以生物质为原料可绿色高效合成饲用氨基酸，而且其具有原料可再生、成本低廉、反应条件温和、环境污染小等优点，为解决动植物提取和化学炼制引起的环境污染问题提供了一种有效解决方案。本文针对饲用氨基酸（赖氨酸、甲硫氨酸、色氨酸、苏氨酸、缬氨酸和精氨酸）的生物合成途径，介绍了大肠杆菌合成饲用氨基酸的生产瓶颈，并从饲用氨基酸大肠杆菌细胞工厂的构建与优化，综述了利用合成生物学技术改造大肠杆菌细胞工厂合成饲用氨基酸的研究现状。提升饲用氨基酸的生产技术水平、提高大肠杆菌细胞的鲁棒性和增强大肠杆菌细胞对不利环境的耐受能力，可以提升饲用氨基酸发酵性能，简化发酵过程控制，降低饲用氨基酸的生产成本，是未来饲用氨基酸生产菌株工程化改造的方向。

关键词: 饲用氨基酸, 大肠杆菌, 微生物细胞工厂, 合成生物学, 代谢工程

Abstract:

With the rapid development of animal husbandry and fishery, it promotes the development of protein for animal feed. Due to the enhancement of people's safety awareness of food additives, there is an urgent need to enhance the supply of sustainable protein for use in animal feed. Because the protein is composed of amino acids, the feed amino acids can replace protein for animal feed, which can provide enough nutrition for the animal's growth. Thus, the feed amino acid is an important product that has attracted a great attention by investigators because of the widely uses in the fields of animal feed supplement and has broad market potential. Using synthetic biology and metabolic engineering strategy, the metabolic pathway and regulatory network of Escherichia coli (E. coli) was designed, engineered, and reconstructed to obtain suitable E. coli cell factories.These factories provide a promising and sustainable alternative for the production of feed amino acid from renewable feedstock, and is attracting great attention as it is an economic and environmentally friendly bioprocessing. E. coli cell factories offer an alternative strategy to replace chemical refinery, animal and plant extraction, reducing the harm to the environment and dependence on natural resources. In the present review, we discussed the biosynthesis pathway of feed amino acid (lysine, methionine, tryptophan, threonine, valine, and arginine) in E. coli. According to the biosynthesis pathway of feed amino acid, the production bottlenecks of feed amino acid in E. coli cell factories were discussed. We also summarized recent studies about in the feed amino acid production from the constructed and optimized of E. coli cell factory. Furthermore, we proposed future research directions to improve the technical level of feed amino acid and enhance the robustness of E. coli cell factories for reducing the cost of bioproduction, simplifying downstream separation, and enhancing industrial productivity.

Key words: feed amino acid, Escherichia coli, microbial cell factories, synthetic biology, metabolic engineering

中图分类号:

郭亮, 高聪, 柳亚迪, 陈修来, 刘立明. 大肠杆菌生产饲用氨基酸的研究进展[J]. 合成生物学, 2021, 2(6): 964-981, doi: 10.12211/2096-8280.2021-042.

Liang GUO, Cong GAO, Yadi LIU, Xiulai CHEN, Liming LIU. Advances in bioproduction of feed amino acid by Escherichia coli[J]. Synthetic Biology Journal, 2021, 2(6): 964-981, doi: 10.12211/2096-8280.2021-042.

参考文献 99

1	刘华阳, 李祥明. 我国疯牛病传染因子研究进展[J]. 饲料工业, 2020, 41(15): 60-64.
	LIU H Y, LI X M. Research progress of infectious factor of BSE in China[J]. Feed Industry, 2020, 41(15): 60-64.
2	KIDD M T, MAYNARD C W, MULLENIX G J. Progress of amino acid nutrition for diet protein reduction in poultry[J]. Journal of Animal Science and Biotechnology, 2021, 12(1): 45-45.
3	CARMO FELIX F K DO, JUNIOR LETTI L A, DE MELO PEREIRA G V, et al. L-lysine production improvement: a review of the state of the art and patent landscape focusing on strain development and fermentation technologies[J]. Critical Reviews in Biotechnology, 2019, 39(8): 1031-1055.
4	KARAU A, GRAYSON I. Amino acids in human and animal nutrition[M]// ZORN H, CZERMAK P. Biotechnology of food and feed additives, 2014: 189-228.
5	赵静. 几种主要饲用氨基酸的营养研究进展[J]. 中国饲料添加剂, 2016(1): 10-13.
	ZHAO J. Research advance on nutritional effects of several major feeding amino acid[J]. China Feed Additive, 2016(1): 10-13.
6	ZHANG Y N, WANG S, DENG Y Z, et al. The application of reduced dietary crude protein levels supplemented with additional amino acids in laying ducks[J]. Poultry Science, 2021, 100(4): 1-7.
7	YANG Z Y, HTOO J K, LIAO S F. Methionine nutrition in swine and related monogastric animals: beyond protein biosynthesis[J]. Animal Feed Science and Technology, 2020, 268: 114608.
8	YOHAN P, JU-HWAN P, SURYEON P, et al. Enhanced cellular uptake and pharmacokinetic characteristics of doxorubicin-valine amide prodrug[J]. Molecules, 2016, 21(10): 1-9.
9	谭伟杰. 2016年中国饲用氨基酸产业透视[J]. 中国畜牧杂志, 2016, 52(8): 12-16.
	TAN W J. The perspective of China feed amino acid industry in 2016[J]. Chinese Journal of Animal Science, 2016, 52(8): 12-16.
10	董德宽. 合成的饲用氨基酸[J]. 国外畜牧科技, 1983(6): 11-12.
	DONG D K. Synthetic feed amino acids[J]. Animal Science Abroad, 1983(6): 11-12.
11	LIU Y N, LI Q, ZHENG P, et al. Developing a high-throughput screening method for threonine overproduction based on an artificial promoter[J]. Microbial Cell Factories, 2015, 14: 1-11.
12	董迅衍, 王小元. 微生物生产L-苏氨酸的代谢工程研究进展[J]. 食品与生物技术学报, 2016(12): 1233-1240.
	DONG X Y, WANG X Y. Advances in microbial metabolic engineering to increase L-threonine production[J]. Journal of Food Science and Biotechnology, 2016, 35(12): 1233-1240.
13	鲁佩玉, 孙青华. 发酵法生产氨基酸工艺研究[J]. 中国调味品, 2019, 44(1): 176-178.
	LU P Y, SUN Q H. Study on the production process of amino acids by fermentation method[J]. China Condiment, 2019, 44(1): 176-178.
14	LIU L, BILAL M, LUO H, et al. Metabolic engineering and fermentation process strategies for L-tryptophan production by Escherichia coli[J]. Processes, 2019, 7(4): 1-17.
15	YANG D, PARK S Y, PARK Y S, et al. Metabolic engineering of Escherichia coli for natural product biosynthesis[J]. Trends in Biotechnology, 2020, 38(7): 745-765.
16	WEI J, WANG Z, YU H, et al. Construction and fermentation of L-threonine-producing recombinant Escherichia coli[J]. Microbiology China, 2019, 46(4): 695-706.
17	HUCCETOGULLARI D, LUO Z W, LEE S Y. Metabolic engineering of microorganisms for production of aromatic compounds[J]. Microbial Cell Factories, 2019, 18(1): 1-29.
18	CHEN X, GAO C, GUO L, et al. DCEO biotechnology: tools to design, construct, evaluate, and optimize the metabolic pathway for biosynthesis of chemicals[J]. Chemical Reviews, 2017, 118(1): 4-72.
19	D'ESTE M, ALVARADO-MORALES M, ANGELIDAKI I. Amino acids production focusing on fermentation technologies-A review[J]. Biotechnology Advances, 2018, 36(1): 14-25.
20	WENDISCH V F. Metabolic engineering advances and prospects for amino acid production[J]. Metabolic Engineering, 2020, 58: 17-34.
21	马倩, 夏利, 谭淼, 等. 氨基酸生产的代谢工程研究进展与发展趋势[J]. 生物工程学报, 2020, 37(5): 1-20.
	MA Q, XIA L, TAN M, et al. Advances and prospects in metabolic engineering for the production of amino acids[J]. Chinese Journal of Biotechnology, 2021, 37(5): 1-20.
22	赵嫚, 彭莉, 成浩, 等. 微生物甲硫氨酸合成调控的综合研究进展与展望[J]. 食品与发酵工业, 2020, 46(24): 257-264.
	ZHAO M, PENG L, CHENG H, et al. Advances on the biosynthesis and regulation of methionine[J]. Food and Fermentation Industries, 2020, 46(24): 257-264.
23	BECKER J, WITTMANN C. A field of dreams: lignin valorization into chemicals, materials, fuels, and health-care products[J]. Biotechnology Advances, 2019, 37(6): 107360.
24	CHEN Z, LIU D. Toward glycerol biorefinery: metabolic engineering for the production of biofuels and chemicals from glycerol[J]. Biotechnology for Biofuels, 2016, 9(1): 205.
25	王昕, 王静, 陈可泉, 等. 合成生物技术制备脂肪族二元胺的研究进展[J]. 合成生物学, 2020, 1(1): 71-83.
	WANG X, WANG J, CHEN K Q, et al. Research progress in bioproduction of aliphatic diamines by synthetic biotechnology[J]. Synthetic Biology Journal, 2020, 1(1): 71-83.
26	TROENDLE J, SCHOPPEL K, BLEIDT A, et al. Metabolic control analysis of L-tryptophan production with Escherichia coli based on data from short-term perturbation experiments[J]. Journal of Biotechnology, 2020, 307: 15-28.
27	DAS A, TYAGI N, VERMA A, et al. Metabolic engineering of Escherichia coli W3110 strain by incorporating genome-level modifications and synthetic plasmid modules to enhance L-Dopa production from glycerol[J]. Preparative Biochemistry and Biotechnology, 2018, 48(8): 671-682.
28	PONTRELLI S, T-Y CHIU, LAN E I, et al. Escherichia coli as a host for metabolic engineering[J]. Metabolic Engineering, 2018, 50: 16-46.
29	LEE J-H, WENDISCH V F. Production of amino acids - Genetic and metabolic engineering approaches[J]. Bioresource Technology, 2017, 245: 1575-1587.
30	LEE S Y, KIM H U, CHAE T U, et al. A comprehensive metabolic map for production of bio-based chemicals[J]. Nature Catalysis, 2019, 2(1): 18-33.
31	YANG Z Y, HASAN M S, HTOO J K, et al. Effects of dietary supplementation of L-methionine vs. DL-methionine on performance, plasma concentrations of free amino acids and other metabolites, and myogenesis gene expression in young growing pigs[J]. Translational Animal Science, 2019, 3(1): 329-339.
32	WANG Y-Y, XU J-Z, ZHANG W-G. Metabolic engineering of L-leucine production in Escherichia coli and Corynebacterium glutamicum: a review[J]. Critical Reviews in Biotechnology, 2019, 39(5): 633-647.
33	MA Q, ZHANG Q, XU Q, et al. Systems metabolic engineering strategies for the production of amino acids[J]. Synthetic and Systems Biotechnology, 2017, 2(2): 87-96.
34	张震, 熊海波, 徐庆阳. 大肠杆菌高密度培养发酵L-色氨酸[J]. 食品与发酵工业, 2019, 45(23): 15-20.
	ZHANG Z, XIONG H B, XU Q Y. L-tryptophan fermentation by high cell density culture of Escherichia coli[J]. Food and Fermentation Industries, 2019, 45(23): 15-20.
35	LI Y, WEI H, WANG T, et al. Current status on metabolic engineering for the production of L-aspartate family amino acids and derivatives[J]. Bioresource Technology, 2017, 245: 1588-1602.
36	FANG Y, WANG J, MA W, et al. Rebalancing microbial carbon distribution for L-threonine maximization using a thermal switch system[J]. Metabolic Engineering, 2020, 61: 33-46.
37	YANG D, YOO S M, GU C, et al. Expanded synthetic small regulatory RNA expression platforms for rapid and multiplex gene expression knockdown[J]. Metabolic Engineering, 2019, 54: 180-190.
38	HUANG J F, LIU Z Q, JIN L Q, et al. Metabolic engineering of Escherichia coli for microbial production of L-methionine[J]. Biotechnology and Bioengineering, 2017, 114(4): 843-851.
39	HUANG J-F, SHEN Z-Y, MAO Q-L, et al. Systematic analysis of bottlenecks in a multibranched and multilevel regulated pathway: the molecular fundamentals of L-methionine biosynthesis in Escherichia coli[J]. ACS Synthetic Biology, 2018, 7(11): 2577-2589.
40	TELEKI A, RAHNERT M, BUNGART O, et al. Robust identification of metabolic control for microbial L-methionine production following an easy-to-use puristic approach[J]. Metabolic Engineering, 2017, 41: 159-172.
41	ZHAO H, FANG Y, WANG X, et al. Increasing L-threonine production in Escherichia coli by engineering the glyoxylate shunt and the L-threonine biosynthesis pathway[J]. Applied Microbiology and Biotechnology, 2018, 102(13): 5505-5518.
42	XU J-Z, YANG H-K, ZHANG W-G. NADPH metabolism: a survey of its theoretical characteristics and manipulation strategies in amino acid biosynthesis[J]. Critical Reviews in Biotechnology, 2018, 38(7): 1061-1076.
43	高虎涛, 申晓林, 孙新晓, 等. 代谢工程调控策略在生物合成氨基酸及其衍生物中的应用[J]. 化工学报, 2020, 71(9): 4058-4070.
	GAO H T, SHEN X L, SUN X X, et al. Metabolic engineering strategies in biosynthesis of amino acids and their derivatives[J]. CIESC Journal, 2020, 71(9): 4058-4070.
44	李华, 董伟, 李由然, 等. 大肠杆菌MetD运输系统缺失对蛋氨酸积累的影响[J]. 微生物学通报, 2017, 44(6): 1416-1426.
	LI H, DONG W, LI Y R, et al. Effect of inactivating MetD transporter system of Escherichia coli W3110 on L-methionine production[J]. Microbiology China, 2017, 44(6): 1416-1426.
45	LIU Q, LIANG Y, ZHANG Y, et al. YjeH Is a novel exporter of L-methionine and branched-chain amino acids in Escherichia coli[J]. Applied and Environmental Microbiology, 2015, 81(22): 7753-7766.
46	杨冬美, 李华, 李由然, 等. 大肠杆菌TdcC、SstT和LIV-1系统缺失对胞外L-苏氨酸积累的影响[J]. 微生物学通报, 2017, 44(1): 20-29.
	YANG D M, LI H, LI Y R, et al. Effects of TdcC、SstT and LIV-1 systems deletion of Escherichia coli on extracellular L-threonine accumulation[J]. Microbiology China, 2017, 44(1): 20-29.
47	YING H X, HE X, LI Y, et al. Optimization of culture conditions for enhanced lysine production using engineered Escherichia coli[J]. Applied Biochemistry and Biotechnology, 2014, 172(8): 3835-3843.
48	XU J, HAN M, REN X, et al. Modification of aspartokinase Ⅲ and dihydrodipicolinate synthetase increases the production of L-lysine in Escherichia coli[J]. Biochemical Engineering Journal, 2016, 114: 79-86.
49	GENG F, CHEN Z, ZHENG P, et al. Exploring the allosteric mechanism of dihydrodipicolinate synthase by reverse engineering of the allosteric inhibitor binding sites and its application for lysine production[J]. Applied Microbiology and Biotechnology, 2013, 97(5): 1963-1971.
50	WANG X, LI Q, SUN C, et al. GREACE-assisted adaptive laboratory evolution in endpoint fermentation broth enhances lysine production by Escherichia coli[J]. Microbial Cell Factories, 2019, 18: 1-13.
51	PIAO X, WANG L, LIN B, et al. Metabolic engineering of Escherichia coli for production of L-aspartate and its derivative beta-alanine with high stoichiometric yield[J]. Metabolic Engineering, 2019, 54: 244-254.
52	WANG S, HOU Y, CHEN X, et al. Kick-starting evolution efficiency with an autonomous evolution mutation system[J]. Metabolic Engineering, 2019, 54: 127-136.
53	YE C, LUO Q, GUO L, et al. Improving lysine production through construction of an Escherichia coli enzyme-constrained model[J]. Biotechnology and Bioengineering, 2020, 117(11): 3533-3544.
54	ZHANG X, YAN J A, YU L, et al. Construction of recombinant plasmids containing threonine operon and their effects on L-threonine accumulation[J]. Acta Microbiologica Sinica, 2009, 49(5): 591-596.
55	LI Y J, ZHANG D Z, CAI N Y, et al. Betaine supplementation improved L-threonine fermentation of Escherichia coli THRD by upregulating zwf (glucose-6-phosphate dehydrogenase) expression[J]. Electronic Journal of Biotechnology, 2019, 39: 67-73.
56	LIU J, LI H, XIONG H, et al. Two-stage carbon distribution and cofactor generation for improving L-threonine production of Escherichia coli[J]. Biotechnology and Bioengineering, 2019, 116(1): 110-120.
57	TANG X-L, DU X-Y, CHEN L-J, et al. Enhanced production of L-methionine in engineered Escherichia coli with efficient supply of one carbon unit[J]. Biotechnology Letters, 2020, 42(3): 429-436.
58	LI H, WANG B S, LI Y R, et al. Metabolic engineering of Escherichia coli W3110 for the production of L-methionine[J]. Journal of Industrial Microbiology and Biotechnology, 2017, 44(1): 75-88.
59	AZUMA S, TSUNEKAWA H, OKABE M, et al. Hyper-production of L-trytophan via fermentation with crystallization[J]. Applied Microbiology and Biotechnology, 1993, 39: 471-476.
60	ZHAO Z-J, ZOU C, ZHU Y-X, et al. Development of L-tryptophan production strains by defined genetic modification in Escherichia coli[J]. Journal of Industrial Microbiology and Biotechnology, 2011, 38(12): 1921-1929.
61	AIBA S, TSUNEKAWA H, IMANAKA T. New approach to tryptophan production by Escherichia coli: genetic manipulation of composite plasmids in vitro[J]. Applied and Environmental Microbiology, 1982, 43(2): 289-297.
62	CHEN Y, LIU Y, DING D, et al. Rational design and analysis of an Escherichia coli strain for high-efficiency tryptophan production[J]. Journal of Industrial Microbiology and Biotechnology, 2018, 45(5): 357-367.
63	DU L, ZHANG Z, XU Q, et al. New strategy for removing acetic acid as a by-product during L-tryptophan production[J]. Biotechnology & Biotechnological Equipment, 2019, 33(1): 1471-1480.
64	PARK J H, LEE K H, KIM T Y, et al. Metabolic engineering of Escherichia coli for the production of L-valine based on transcriptome analysis and in silico gene knockout simulation[J]. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(19): 7797-7802.
65	PARK J H, KIM T Y, LEE K H, et al. Fed-batch culture of Escherichia coli for L-valine production based on in silico flux response analysis[J]. Biotechnology and Bioengineering, 2011, 108(4): 934-946.
66	SAVRASOVA E A, STOYNOVA N V. Application of leucine dehydrogenase Bcd from Bacillus subtilis for L-valine synthesis in Escherichia coli under microaerobic conditions[J]. Heliyon, 2019, 5(4): e01406.
67	HAO Y, MA Q, LIU X, et al. High-yield production of L-valine in engineered Escherichia coli by a novel two-stage fermentation[J]. Metabolic Engineering, 2020, 62: 198-206.
68	XU M, RAO Z, YANG J, et al. Heterologous and homologous expression of the arginine biosynthetic argC similar to H cluster from Corynebacterium crenatum for improvement of L-arginine production[J]. Journal of Industrial Microbiology and Biotechnology, 2012, 39(3): 495-502.
69	GINESY M, BELOTSERKOVSKY J, ENMAN J, et al. Metabolic engineering of Escherichia coli for enhanced arginine biosynthesis[J]. Microbial Cell Factories, 2015, 14: 1-11.
70	SANDER T, WANG C Y, GLATTER T, et al. CRISPRi-based downregulation of transcriptional feedback improves growth and metabolism of arginine overproducing E. coli[J]. ACS Synthetic Biology, 2019, 8(9): 1983-1990.
71	LEE K H, PARK J H, KIM T Y, et al. Systems metabolic engineering of Escherichia coli for L-threonine production[J]. Molecular Systems Biology, 2007, 3:149.
72	CHEN L, CHEN M, MA C, et al. Discovery of feed-forward regulation in L-tryptophan biosynthesis and its use in metabolic engineering of E. coli for efficient tryptophan bioproduction[J]. Metabolic Engineering, 2018, 47: 434-444.
73	LI M, LIU C, YANG J, et al. Common problems associated with the microbial productions of aromatic compounds and corresponding metabolic engineering strategies[J]. Biotechnology Advances, 2020, 41: 107548.
74	JIANG J, LIU T, LIN S. Research progress on the biosynthesis of aromatic compounds by microorganisms[J]. Chinese Bulletin of Life Sciences, 2019, 31(5): 430-448.
75	LI Z, WANG H, DING D, et al. Metabolic engineering of Escherichia coli for production of chemicals derived from the shikimate pathway[J]. Journal of Industrial Microbiology and Biotechnology, 2020, 47(6-7): 525-535.
76	CHEN L, ZENG A-P. Rational design and metabolic analysis of Escherichia coli for effective production of L-tryptophan at high concentration[J]. Applied Microbiology and Biotechnology, 2017, 101(2): 559-568.
77	TROENDLE J, TRACHTMANN N, SPRENGER G A, et al. Fed-batch production of L-tryptophan from glycerol using recombinant Escherichia coli[J]. Biotechnology and Bioengineering, 2018, 115(12): 2881-2892.
78	NIU H, LI R, LIANG Q, et al. Metabolic engineering for improving L-tryptophan production in Escherichia coli[J]. Journal of Industrial Microbiology and Biotechnology, 2019, 46(1): 55-65.
79	XIONG B, ZHU Y, TIAN D, et al. Flux redistribution of central carbon metabolism for efficient production of L-tryptophan in Escherichia coli[J]. Biotechnology and Bioengineering, 2021, 118(3): 1393-1404.
80	LEE J, KIM J, SONG J E, et al. Production of Tyrian purple indigoid dye from tryptophan in Escherichia coli[J]. Nature Chemical Biology, 2020: 1-9.
81	WYNANDS B, LENZEN C, OTTO M, et al. Metabolic engineering of Pseudomonas taiwanensis VLB120 with minimal genomic modifications for high-yield phenol production[J]. Metabolic Engineering, 2018, 47: 121-133.
82	DING R, LIU L, CHEN X, et al. Introduction of two mutations into AroG increases phenylalanine production in Escherichia coli[J]. Biotechnology Letters, 2014, 36(10): 2103-2108.
83	NODA S, KONDO A. Recent advances in microbial production of aromatic chemicals and derivatives[J]. Trends in Biotechnology, 2017, 35(8): 785-796.
84	WANG J, ZHANG R, ZHANG Y, et al. Developing a pyruvate-driven metabolic scenario for growth-coupled microbial production[J]. Metabolic Engineering, 2019, 55: 191-200.
85	GUO L, ZHANG F, ZHANG C, et al. Enhancement of malate production through engineering of the periplasmic rTCA pathway in Escherichia coli[J]. Biotechnology and Bioengineering, 2018, 115(6): 1571-1580.
86	LIU L, CHEN S, WU J. Phosphoenolpyruvate: glucose phosphotransferase system modification increases the conversion rate during L-tryptophan production in Escherichia coli[J]. Journal of Industrial Microbiology and Biotechnology, 2017, 44(10): 1385-1395.
87	AL-BASSAM M M, KIM J N, ZARAMELA L S, et al. Optimization of carbon and energy utilization through differential translational efficiency[J]. Nature Communications, 2018, 9(1): 1-13.
88	KOENDJBIHARIE J G, KRANENBURG R VAN, KENGEN S W M. The PEP-pyruvate-oxaloacetate node: variation at the heart of metabolism[J]. FEMS Microbiology Reviews, 2020: 1-19.
89	DIAO J, SONG X, ZHANG L, et al. Tailoring cyanobacteria as a new platform for highly efficient synthesis of astaxanthin[J]. Metabolic Engineering, 2020, 61: 275-287.
90	NGUYEN A D, KIM D, LEE E Y. Unlocking the biosynthesis of sesquiterpenoids from methane via the methylerythritol phosphate pathway in methanotrophic bacteria, using alpha-humulene as a model compound[J]. Metabolic Engineering, 2020, 61: 69-78.
91	王均成, 王可, 张春宇. L-缬氨酸的应用和育种研究进展[J]. 发酵科技通讯, 2012, 41(01): 30-34.
	WANG J C, WANG K, ZHANG C Y. Advances in bioproduction of L-valine[J]. Bulletin of Fermentation Science and Technology, 2012, 41(1): 30-34.
92	JONES C M, LOZADA N J H, PFLEGER B F. Efflux systems in bacteria and their metabolic engineering applications[J]. Applied Microbiology and Biotechnology, 2015, 99(22): 9381-9393.
93	CHARLIER D, BERVOETS I. Regulation of arginine biosynthesis, catabolism and transport in Escherichia coli[J]. Amino Acids, 2019, 51(8): 1103-1127.
94	程功, 徐建中, 张伟国. L-精氨酸生物合成机制及其代谢工程育种研究进展[J]. 微生物学通报, 2016, 43(6): 1379-1387.
	CHENG G, XU J Z, ZHANG W G. Progress in biosynthesis and metabolic engineering of L-arginine producer[J]. Microbiology China, 2016, 43(6): 1379-1387.
95	LU C D. Pathways and regulation of bacterial arginine metabolism and perspectives for obtaining arginine overproducing strains[J]. Applied Microbiology and Biotechnology, 2006, 70(3): 261-272.
96	CHARLIER D, ROOVERS M, VLIET F VAN, et al. Arginine regulon of Escherichia coli K-12: a study of repressor-operator interactions and of in vitro binding affinities versus in vivo repression[J]. Journal of Molecular Biology, 1992, 226(2): 367-386.
97	NANDINENI M R, GOWRISHANKAR J. Evidence for an arginine exporter encoded by yggA (argO) that is regulated by the LysR-type transcriptional regulator ArgP in Escherichia coli[J]. Journal of Bacteriology, 2004, 186(11): 3539-3546.
98	RAJAGOPAL B S, DEPONTE J, TUCHMAN M, et al. Use of inducible feedback-resistant N-acetylglutamate synthetase (argA) genes for enhanced arginine biosynthesis by genetically engineered Escherichia coli K-12 strains[J]. Applied and Environmental Microbiology, 1998, 64(5): 1805-1811.
99	GINESY M, RUSANOVA-NAYDENOVA D, ROVA U. Tuning of the carbon-to-nitrogen ratio for the production of L-arginine by Escherichia coli[J]. Fermentation, 2017, 3(4): 1-11.

[1]	卞佳豪, 杨广宇. 人工智能辅助的蛋白质工程[J]. 合成生物学, 2022, 3(3): 429-444.
[2]	冯晴晴, 张天鲛, 赵潇, 聂广军. 合成纳米生物学——合成生物学与纳米生物学的交叉前沿[J]. 合成生物学, 2022, 3(2): 260-278.
[3]	施茜, 吴园园, 杨洋. DNA纳米技术与合成生物学[J]. 合成生物学, 2022, 3(2): 302-319.
[4]	胥欣欣, 匡华. 基于合成受体的食品污染物生物检测进展[J]. 合成生物学, 2022, 3(2): 399-414.
[5]	武伟红, 李炜, 张先恩, 崔宗强. 合成生物学与荧光成像技术[J]. 合成生物学, 2022, 3(2): 369-384.
[6]	郑涵奇, 吴晴, 李洪军, 顾臻. 合成生物学与纳米生物学的交叉融合及其在生物医药领域的应用[J]. 合成生物学, 2022, 3(2): 279-301.
[7]	张亭, 冷梦甜, 金帆, 袁海. 合成生物研究重大科技基础设施概述[J]. 合成生物学, 2022, 3(1): 184-194.
[8]	郭姝媛, 吴良焕, 刘香健, 王博, 于涛. 微生物中一碳代谢网络构建的进展与挑战[J]. 合成生物学, 2022, 3(1): 116-137.
[9]	赵晓宇, 张浩, 李雪飞, 胡政. 进化视角下的定量生物学规律与人工生命合成[J]. 合成生物学, 2022, 3(1): 6-21.
[10]	褚亚东, 赵宗保. 小型集成化自动移液工作站系统及应用[J]. 合成生物学, 2022, 3(1): 195-208.
[11]	郭思敏, 叶斌, 徐飞. 美德伦理视角下的合成生物学技术伦理治理[J]. 合成生物学, 2022, 3(1): 224-237.
[12]	任师超, 孙秋艳, 冯旭东, 李春. 微生物细胞工厂合成五环三萜皂苷类化合物[J]. 合成生物学, 2022, 3(1): 168-183.
[13]	陈久洲, 王钰, 蒲伟, 郑平, 孙际宾. 5-氨基乙酰丙酸生物合成技术的发展及展望[J]. 合成生物学, 2021, 2(6): 1000-1016.
[14]	严伟, 高豪, 蒋羽佳, 钱秀娟, 周杰, 董维亮, 章文明, 信丰学, 姜岷. 2-苯乙醇生物合成的研究进展[J]. 合成生物学, 2021, 2(6): 1030-1045.
[15]	汪庆卓, 宋萍, 黄和. 合成生物技术驱动天然的真核油脂细胞工厂开发[J]. 合成生物学, 2021, 2(6): 920-941.