天冬氨酸族饲用氨基酸微生物细胞工厂的创制

doi:10.12211/2096-8280.2025-032

摘要/Abstract

摘要：

氨基酸作为动物饲料的重要组成部分，是提高畜禽消化机能、禽肉品质、蛋白转化效率以及降低豆粕使用量的关键要素。合成生物技术的快速发展为氨基酸高产菌株构建和优化铺平了道路，极大地提升了氨基酸生产效率，显著降低了生产成本。本文在分析L-赖氨酸、L-甲硫氨酸、L-苏氨酸和L-异亮氨酸等四种天冬氨酸族氨基酸合成途径的基础上，详细介绍了菌种改造方法和策略，包括代谢路径重构、代谢途径优化、辅因子供应和增强产物外排等四个方面，未来要从工业环境抗逆性、底物利用范围的拓展以及动态调控系统的优化三个方面进行突破，才能为高性能氨基酸生产菌株的创制提供理论指导和技术支撑。

关键词: 天冬氨酸族氨基酸, 微生物细胞工厂, 代谢工程, 合成生物学, 基因编辑

Abstract:

Amino acids are essential components of animal feed, playing key roles in improving digestive function in livestock, enhancing meat quality, increasing protein conversion efficiency, and reducing reliance on soybean meal. Driven by global population growth and dietary changes, the increasing demand for animal protein has strained the livestock industry. This industry traditionally relies heavily on soybean meal as its primary protein source, a method that results in low nitrogen utilization and exacerbates environmental pollution from nitrogen emissions. Aspartate-family amino acids, including L-lysine, L-methionine, L-threonine, and L-isoleucine, represent the most significant category of feed amino acids, accounting for nearly 90% of global consumption. They address current challenges by balancing feed nutrition according to the ideal protein standard and enabling a low-carbon transition in animal husbandry. The primary method for producing these four amino acids is through microbial fermentation, with Escherichia coli and Corynebacterium glutamicum serving as the primary host organisms. Rapid advances in systems biology, synthetic biology, metabolic engineering, and evolutionary engineering have facilitated the construction and optimization of high-yield amino acid-producing strains. This has significantly enhanced production efficiency and substantially reduced costs. Based on an analysis of the aspartate-family amino acid biosynthetic pathways, this paper details strain modification methods and strategies. These encompass four key aspects: metabolic pathway reconstruction, metabolic pathway optimization, cofactor supply enhancement, and improved product efflux. These approaches have enabled the industrial-scale production of strains achieving high titers and yields. Finally, future research directions are discussed, focusing on three fronts: enhancing strain stress resistance in industrial environments, expanding the range of utilizable substrates, and optimizing dynamic regulatory systems. These advancements are intended to offer theoretical guidance and technical support for the development of high-performance amino acid-producing strains. The ultimate objective is to facilitate the global shift towards efficient and environmentally sustainable feed amino acid production, thereby alleviating pressures on protein resources.

Key words: aspartate-family amino acids, microbial cell factory, metabolic engineering, synthetic biology, gene editing

中图分类号:

Q815

赵欣雨, 盛琦, 刘开放, 刘佳, 刘立明. 天冬氨酸族饲用氨基酸微生物细胞工厂的创制[J]. 合成生物学, 2025, 6(5): 1184-1202.

ZHAO Xinyu, SHENG Qi, LIU Kaifang, LIU Jia, LIU Liming. Construction of microbial cell factories for aspartate-family feed amino acids[J]. Synthetic Biology Journal, 2025, 6(5): 1184-1202.

图/表 10

参考文献 30

[31]	ZHANG Q Q, WANG Y H, WANG X L, et al. Metabolic engineering of Escherichia coli for efficient L-isoleucine production based on the citramalate pathway[J]. Journal of Agricultural and Food Chemistry, 2025, 73(19): 11900-11911.
[32]	GUILLOUET S, RODAL A, AN G H, et al. Metabolic redirection of carbon flow toward isoleucine by expressing a catabolic threonine dehydratase in a threonine-overproducing Corynebacterium glutamicum [J]. Applied Microbiology and Biotechnology, 2001, 57(5): 667-673.
[33]	YU S Z, ZHENG B, CHEN Z Y, et al. Metabolic engineering of Corynebacterium glutamicum for producing branched chain amino acids[J]. Microbial Cell Factories, 2021, 20(1): 230.
[34]	GUO Y F, HAN M, XU J Z, et al. Analysis of acetohydroxyacid synthase variants from branched-chain amino acids-producing strains and their effects on the synthesis of branched-chain amino acids in Corynebacterium glutamicum [J]. Protein Expression and Purification, 2015, 109: 106-112.
[35]	YIN L H, HU X Q, XU D Q, et al. Co-expression of feedback-resistant threonine dehydratase and acetohydroxy acid synthase increase L-isoleucine production in Corynebacterium glutamicum [J]. Metabolic Engineering, 2012, 14(5): 542-550.
[36]	SHI J, FENG Z Z, SONG Q, et al. Structural and functional insights into transcription activation of the essential LysR-type transcriptional regulators[J]. Protein Science, 2024, 33(6): e5012.
[37]	DE GRAAF A A, EGGELING L, SAHM H. Metabolic engineering for L-lysine production by Corynebacterium glutamicum [J]. Advances in Biochemical Engineering/Biotechnology, 2001, 73: 9-29.
[38]	GRUZDEV N, HACHAM Y, HAVIV H, et al. Conversion of methionine biosynthesis in Escherichia coli from trans- to direct-sulfurylation enhances extracellular methionine levels[J]. Microbial Cell Factories, 2023, 22(1): 151.
[39]	ZHOU Z, ZHANG X Y, WU J, et al. Targeting cofactors regeneration in methylation and hydroxylation for high level production of Ferulic acid[J]. Metabolic Engineering, 2022, 73: 247-255.
[40]	RHEE K Y, PAREKH B S, HATFIELD G W. Leucine-responsive regulatory protein-DNA interactions in the leader region of the ilvGMEDA operon of Escherichia coli [J]. Journal of Biological Chemistry, 1996, 271(43): 26499-26507.
[41]	JAFRI S, CHEN S L, CALVO J M. ilvIH operon expression in Escherichia coli requires Lrp binding to two distinct regions of DNA[J]. Journal of Bacteriology, 2002, 184(19): 5293-5300.
[42]	MORBACH S, JUNGER C, SAHM H, et al. Attenuation control of ilvBNC in Corynebacterium glutamicum: evidence of leader peptide formation without the presence of a ribosome binding site[J]. Journal of Bioscience and Bioengineering, 2000, 90(5): 501-507.
[1]	FANG Q C, ZHANG X Y, DAI G C, et al. Low-opportunity-cost feed can reduce land-use-related environmental impacts by about one-third in China[J]. Nature Food, 2023, 4(8): 677-685.
[2]	MA Q, ZHANG Q W, XU Q Y, et al. Systems metabolic engineering strategies for the production of amino acids[J]. Synthetic and Systems Biotechnology, 2017, 2(2): 87-96.
[3]	刘佳, 盛琦, 刘开放, 等. 微生物制造饲用氨基酸助力豆粕减量替代[J]. 中国科学院院刊, 2025, 40(1): 25-35.
	LIU J, SHENG Q, LIU K F, et al. Microbial production of feed amino acids promotes reduction and replacement of soybean meal[J]. Bulletin of Chinese Academy of Sciences, 2025, 40(1): 25-35.
[4]	KIM S Y. 9 What would be the next feed amino acid based on a microbial point of view?[J]. Journal of Animal Science, 2021, 99(S1): 12-13.
[5]	LIAO J L, ZHANG P G, YIN J D, et al. New insights into the effects of dietary amino acid composition on meat quality in pigs: a review[J]. Meat Science, 2025, 221: 109721.
[6]	OLUWABIYI C T, SONG Z G. Branched-chain amino acids supplementation in low-protein broiler diets: a review[J]. Animal Feed Science and Technology, 2024, 318: 116114.
[7]	LI M H, LI H, ZHANG X, et al. Metabolic engineering of Corynebacterium glutamicum: unlocking its potential as a key cell factory platform for organic acid production[J]. Biotechnology Advances, 2024, 77: 108475.
[8]	LIU J, XU J Z, RAO Z M, et al. Industrial production of L-lysine in Corynebacterium glutamicum: progress and prospects[J]. Microbiological Research, 2022, 262: 127101.
[9]	LI C Y, ZHANG R H, WANG J, et al. Protein engineering for improving and diversifying natural product biosynthesis[J]. Trends in Biotechnology, 2020, 38(7): 729-744.
[10]	DONG X Y, ZHAO Y, HU J Y, et al. Attenuating L-lysine production by deletion of ddh and lysE and their effect on L-threonine and L-isoleucine production in Corynebacterium glutamicum [J]. Enzyme and Microbial Technology, 2016, 93-94: 70-78.
[11]	BOMMAREDDY R R, CHEN Z, RAPPERT S, et al. A de novo NADPH generation pathway for improving lysine production of Corynebacterium glutamicum by rational design of the coenzyme specificity of glyceraldehyde 3-phosphate dehydrogenase[J]. Metabolic Engineering, 2014, 25: 30-37.
[43]	BOOB A G, CHEN J Y, ZHAO H M. Enabling pathway design by multiplex experimentation and machine learning[J]. Metabolic Engineering, 2024, 81: 70-87.
[44]	DING D Q, ZHU Y R, BAI D Y, et al. Monitoring and dynamically controlling glucose uptake rate and central metabolism[J]. Nature Chemical Engineering, 2025, 2(1): 50-62.
[45]	SHENG Q, YI L X, ZHONG B, et al. Shikimic acid biosynthesis in microorganisms: current status and future direction[J]. Biotechnology Advances, 2023, 62: 108073.
[46]	XU J Z, HAN M, REN X D, 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.
[47]	LIU J H, LIU J, LI J J, et al. Reconstruction the feedback regulation of amino acid metabolism to develop a non-auxotrophic L-threonine producing Corynebacterium glutamicum [J]. Bioresources and Bioprocessing, 2024, 11(1): 43.
[48]	LEE J H, LEE D E, LEE B U, et al. Global analyses of transcriptomes and proteomes of a parent strain and an L-threonine-overproducing mutant strain[J]. Journal of Bacteriology, 2003, 185(18): 5442-5451.
[49]	ZHAO Z Q, YOU J J, SHI X P, et al. Engineering Escherichia coli for L-threonine hyperproduction based on multidimensional optimization strategies[J]. Journal of Agricultural and Food Chemistry, 2024, 72(41): 22682-22691.
[50]	PARK J H, OH J E, LEE K H, et al. Rational design of Escherichia coli for L-isoleucine production[J]. ACS Synthetic Biology, 2012, 1(11): 532-540.
[51]	ZHANG Y C, LIU Y D, ZHANG S Y, et al. Metabolic engineering of Corynebacterium glutamicum WM001 to improve L-isoleucine production[J]. Biotechnology and Applied Biochemistry, 2021, 68(3): 568-584.
[52]	LIU Y D, LI Y Y, WANG X Y. Acetohydroxyacid synthases: evolution, structure, and function[J]. Applied Microbiology and Biotechnology, 2016, 100(20): 8633-8649.
[53]	DUAN M, CHEN S, LIU X L, et al. The application of Corynebacterium glutamicum in L-threonine biosynthesis[J]. Fermentation, 2023, 9(9): 822.
[54]	JIN X, WANG S M, GAO Y P, et al. Combinatorial metabolic engineering of Escherichia coli to efficiently produce L-threonine from untreated cane molasses[J]. Bioresource Technology, 2025, 419: 132058.
[12]	CHEN Y Y, HUANG L G, YU T, et al. Balancing the AspC and AspA pathways of Escherichia coli by systematic metabolic engineering strategy for high-efficient L-homoserine production[J]. ACS Synthetic Biology, 2024, 13(8): 2457-2469.
[13]	BASSALO M C, GARST A D, CHOUDHURY A, et al. Deep scanning lysine metabolism in Escherichia coli [J]. Molecular Systems Biology, 2018, 14(11): e8371.
[14]	OSIRE T, YANG T W, XU M J, et al. Integrated gene engineering synergistically improved substrate-product transport, cofactor generation and gene translation for cadaverine biosynthesis in E. coli [J]. International Journal of Biological Macromolecules, 2021, 169: 8-17.
[15]	XIAO J, WANG D T, WANG L, et al. Increasing L-lysine production in Corynebacterium glutamicum by engineering amino acid transporters[J]. Amino Acids, 2020, 52(10): 1363-1374.
[16]	LIU J, OU Y, XU J Z, et al. L-lysine production by systems metabolic engineering of an NADPH auto-regulated Corynebacterium glutamicum [J]. Bioresource Technology, 2023, 387: 129701.
[17]	MALLA S, VAN DER HELM E, DARBANI B, et al. A novel efficient L-lysine exporter identified by functional metagenomics[J]. Frontiers in Microbiology, 2022, 13: 855736.
[18]	LI Z C, LIU Q, SUN J H, et al. Multivariate modular metabolic engineering for enhanced L-methionine biosynthesis in Escherichia coli [J]. Biotechnology for Biofuels and Bioproducts, 2023, 16(1): 101.
[19]	LI Y, CONG H, LIU B N, et al. Metabolic engineering of Corynebacterium glutamicum for methionine production by removing feedback inhibition and increasing NADPH level[J]. Antonie Van Leeuwenhoek, 2016, 109(9): 1185-1197.
[20]	ZHOU B Z, ZHAO G H, YU J, et al. Multi-step metabolic engineering Corynebacterium glutamicum ATCC13032 to produce L-methionine[J]. Systems Microbiology and Biomanufacturing, 2025, 5(2): 593-610.
[21]	ZHAO Z Q, YOU J J, SHI X P, et al. Multi-module engineering to guide the development of an efficient L-threonine-producing cell factory[J]. Bioresource Technology, 2025, 416: 131802.
[22]	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.
[23]	VO T M, PARK J Y, KIM D, et al. Use of acetate as substrate for sustainable production of homoserine and threonine by Escherichia coli W3110: a modular metabolic engineering approach[J]. Metabolic Engineering, 2024, 84: 13-22.
[55]	YE C, LUO Q L, 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.
[56]	LV Q L, HU M K, TIAN L Z, et al. Enhancing L-glutamine production in Corynebacterium glutamicum by rational metabolic engineering combined with a two-stage pH control strategy[J]. Bioresource Technology, 2021, 341: 125799.
[57]	WANG L J, GUO Y Y, LI M Y, et al. Antibiotic-free high-level L-methionine production in engineered Escherichia coli [J]. Journal of Agricultural and Food Chemistry, 2024, 72(46): 25791-25800.
[58]	JIANG S, WANG R R, WANG D H, et al. Metabolic reprogramming and biosensor-assisted mutagenesis screening for high-level production of L-arginine in Escherichia coli [J]. Metabolic Engineering, 2023, 76: 146-157.
[59]	YE J W, HU D K, CHE X M, et al. Engineering of Halomonas bluephagenesis for low cost production of poly (3-hydroxybutyrate-co-4-hydroxybutyrate) from glucose[J]. Metabolic Engineering, 2018, 47: 143-152.
[60]	YE J W, HU D K, YIN J, et al. Stimulus response-based fine-tuning of polyhydroxyalkanoate pathway in Halomonas [J]. Metabolic Engineering, 2020, 57: 85-95.
[61]	LI T, YE J W, SHEN R, et al. Semirational approach for ultrahigh poly(3-hydroxybutyrate) accumulation in Escherichia coli by combining one-step library construction and high-throughput screening[J]. ACS Synthetic Biology, 2016, 5(11): 1308-1317.
[62]	SHEN R, YIN J, YE J W, et al. Promoter engineering for enhanced P(3HB-co-4HB) production by Halomonas bluephagenesis [J]. ACS Synthetic Biology, 2018, 7(8): 1897-1906.
[63]	DU F, LI Z J, LI X, et al. Optimizing multicopy chromosomal integration for stable high-performing strains[J]. Nature Chemical Biology, 2024, 20(12): 1670-1679.
[64]	BAEK J M, MAZUMDAR S, LEE S W, et al. Butyrate production in engineered Escherichia coli with synthetic scaffolds[J]. Biotechnology and Bioengineering, 2013, 110(10): 2790-2794.
[65]	DUEBER J E, WU G C, MALMIRCHEGINI G R, et al. Synthetic protein scaffolds provide modular control over metabolic flux[J]. Nature Biotechnology, 2009, 27(8): 753-759.
[66]	ZHOU H, VONK B, ROUBOS J A, et al. Algorithmic co-optimization of genetic constructs and growth conditions: application to 6-ACA, a potential nylon-6 precursor[J]. Nucleic Acids Research, 2015, 43(21): 10560-10570.
[67]	VENAYAK N, ANESIADIS N, CLUETT W R, et al. Engineering metabolism through dynamic control[J]. Current Opinion in Biotechnology, 2015, 34: 142-152.
[68]	CRESS B F, TRANTAS E A, VERVERIDIS F, et al. Sensitive cells: enabling tools for static and dynamic control of microbial metabolic pathways[J]. Current Opinion in Biotechnology, 2015, 36: 205-214.
[69]	IMMETHUN C M, DELORENZO D M, FOCHT C M, et al. Physical, chemical, and metabolic state sensors expand the synthetic biology toolbox for Synechocystis sp. PCC 6803[J]. Biotechnology and Bioengineering, 2017, 114(7): 1561-1569.
[70]	SHEN X L, WANG J, LI C Y, et al. Dynamic gene expression engineering as a tool in pathway engineering[J]. Current Opinion in Biotechnology, 2019, 59: 122-129.
[71]	LALWANI M A, ZHAO E M, AVALOS J L. Current and future modalities of dynamic control in metabolic engineering[J]. Current Opinion in Biotechnology, 2018, 52: 56-65.
[72]	PINTO D, VECCHIONE S, WU H, et al. Engineering orthogonal synthetic timer circuits based on extracytoplasmic function σ factors[J]. Nucleic Acids Research, 2018, 46(14): 7450-7464.
[73]	MEYER A J, SEGALL-SHAPIRO T H, GLASSEY E, et al. Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors[J]. Nature Chemical Biology, 2019, 15(2): 196-204.
[74]	叶健文, 陈江楠, 张旭, 等. 动态调控: 一种高效的细胞工厂工程化代谢改造策略[J]. 生物技术通报, 2020, 36(6): 1-12.
	YE J W, CHEN J N, ZHANG X, et al. Dynamic control: an efficient strategy for metabolically engineering microbial cell factories[J]. Biotechnology Bulletin, 2020, 36(6): 1-12.
[75]	BORKOWSKI O, BRICIO C, MURGIANO M, et al. Cell-free prediction of protein expression costs for growing cells[J]. Nature Communications, 2018, 9: 1457.
[76]	GUPTA A, REIZMAN I M B, REISCH C R, et al. Dynamic regulation of metabolic flux in engineered bacteria using a pathway-independent quorum-sensing circuit[J]. Nature Biotechnology, 2017, 35(3): 273-279.
[77]	DING X H, YANG W J, DU X B, et al. High-level and-yield production of L-leucine in engineered Escherichia coli by multistep metabolic engineering[J]. Metabolic Engineering, 2023, 78: 128-136.
[78]	YANG L Y, MU X Q, NIE Y, et al. Improving the production of NAD⁺ via multi-strategy metabolic engineering in Escherichia coli [J]. Metabolic Engineering, 2021, 64: 122-133.
[79]	BOUZON M, DÖRING V, DUBOIS I, et al. Change in cofactor specificity of oxidoreductases by adaptive evolution of an Escherichia coli NADPH-auxotrophic strain[J]. mBio, 2021, 12(4): e0032921.
[80]	XU J Z, RUAN H Z, YU H B, et al. Metabolic engineering of carbohydrate metabolism systems in Corynebacterium glutamicum for improving the efficiency of L-lysine production from mixed sugar[J]. Microbial Cell Factories, 2020, 19(1): 39.
[81]	WANG K, SONG X T, CUI B Y, et al. Metabolic engineering of Escherichia coli for efficient production of ectoine[J]. Journal of Agricultural and Food Chemistry, 2025, 73(1): 646-654.
[82]	LIU B N, SUN X Y, LIU Y, et al. Increased NADPH supply enhances glycolysis metabolic flux and L-methionine production in Corynebacterium glutamicum [J]. Foods, 2022, 11(7): 1031.
[83]	YU F, ZHAO X R, ZHOU J W, et al. Biosynthesis of high-active hemoproteins by the efficient heme-supply Pichia pastoris chassis[J]. Advanced Science, 2023, 10(30): 2302826.
[84]	WANG Y S, BAI Y L, ZENG Q, et al. Recent advances in the metabolic engineering and physiological opportunities for microbial synthesis of L-aspartic acid family amino acids: a review[J]. International Journal of Biological Macromolecules, 2023, 253: 126916.
[85]	赵阔, 程金宇, 郭亮, 等. 谷氨酸棒杆菌代谢工程高效生产L-缬氨酸[J]. 生物工程学报, 2023, 39(8): 3253-3272.
	ZHAO K, CHENG J Y, GUO L, et al. Highly efficient production of L-valine by multiplex metabolic engineering of Corynebacterium glutamicum [J]. Chinese Journal of Biotechnology, 2023, 39(8): 3253-3272.
[86]	XU S Y, ZHOU L, XU Y, et al. Recent advances in structure-based enzyme engineering for functional reconstruction[J]. Biotechnology and Bioengineering, 2023, 120(12): 3427-3445.
[87]	贾男, 臧国伟, 李春, 等. 辅因子在微生物细胞工厂中的代谢调控与应用[J]. 中国生物工程杂志, 2022, 42(7): 79-89.
	JIA N, ZANG G W, LI C, et al. Metabolic regulations and applications of cofactors in microbial cell factories[J]. China Biotechnology, 2022, 42(7): 79-89.
[88]	陈修来, 刘佳, 罗秋玲, 等. 微生物辅因子平衡的代谢调控[J]. 生物工程学报, 2017, 33(1): 16-26.
	CHEN X L, LIU J, LUO Q L, et al. Manipulation of cofactor balance in microorganisms[J]. Chinese Journal of Biotechnology, 2017, 33(1): 16-26.
[89]	陈雅维. ATP调控策略及其在微生物代谢产物合成中的应用[J]. 生物工程学报, 2020, 36(8): 1515-1527.
	CHEN Y W. ATP regulation strategy and its application in the synthesis of microbial metabolites[J]. Chinese Journal of Biotechnology, 2020, 36(8): 1515-1527.
[90]	WEUSTHUIS R A, FOLCH P L, POZO-RODRÍGUEZ A, et al. Applying non-canonical redox cofactors in fermentation processes[J]. iScience, 2020, 23(9): 101471.
[91]	张鸿伟, 王鹏超. 微生物辅因子工程研究进展[J]. 中国生物工程杂志, 2023, 43(4): 112-122.
	ZHANG H W, WANG P C. Research progress of microbial cofactor engineering[J]. China Biotechnology, 2023, 43(4): 112-122.
[92]	GAO C, SONG W, YE C, et al. Bifunctional optogenetic switch powered NADPH availability for improving L-valine production in Escherichia coli [J]. ACS Sustainable Chemistry & Engineering, 2024, 12(41): 15103-15113.
[93]	刘开放. 新型微生物辅因子系统的开发与应用[D]. 无锡: 江南大学, 2024.
	LIU K F. Development and application of novel cofactor systems in microorganisms[D]. Wuxi: Jiangnan University, 2024.
[94]	BLACK W B, ZHANG L Y, MAK W S, et al. Engineering a nicotinamide mononucleotide redox cofactor system for biocatalysis[J]. Nature Chemical Biology, 2020, 16(1): 87-94.
[95]	于袁欢, 周阳, 王欣怡, 等. 光遗传学照进生物医学研究进展[J]. 合成生物学, 2023, 4(1): 102-140.
	YU Y H, ZHOU Y, WANG X Y, et al. Advances in optogenetics for biomedical research[J]. Synthetic Biology Journal, 2023, 4(1): 102-140.
[96]	CHEN R B, YANG S, ZHANG L, et al. Advanced strategies for production of natural products in yeast[J]. iScience, 2020, 23(3): 100879.
[97]	CHEN R B, GAO J Q, YU W, et al. Engineering cofactor supply and recycling to drive phenolic acid biosynthesis in yeast[J]. Nature Chemical Biology, 2022, 18(5): 520-529.
[98]	ZHANG Y F, ZHANG G Y, ZHANG H F, et al. Efficient fermentative production of β-alanine from glucose through multidimensional engineering of Escherichia coli [J]. Journal of Agricultural and Food Chemistry, 2024, 72(25): 14274-14283.
[99]	LINDNER S N, PETROV D P, HAGMANN C T, et al. Phosphotransferase system-mediated glucose uptake is repressed in phosphoglucoisomerase-deficient Corynebacterium glutamicum strains[J]. Applied and Environmental Microbiology, 2013, 79(8): 2588-2595.
[100]	MOLINA-VÁZQUEZ E R, CASPETA L, GOSSET G, et al. Tailoring Escherichia coli BL21 (DE3) for preferential xylose utilization via metabolic and regulatory engineering[J]. Applied Microbiology and Biotechnology, 2025, 109(1): 54.
[101]	ZHU X N, FAN F Y, QIU H N, et al. New xylose transporters support the simultaneous consumption of glucose and xylose in Escherichia coli [J]. mLife, 2022, 1(2): 156-170.
[102]	LI J, QIU Z T, ZHAO G R. Modular engineering of E. coli coculture for efficient production of resveratrol from glucose and arabinose mixture[J]. Synthetic and Systems Biotechnology, 2022, 7(2): 718-729.
[103]	HALLE L, HÖPPNER D, DOSER M, et al. From molasses to purified α-ketoglutarate with engineered Corynebacterium glutamicum [J]. Bioresource Technology, 2025, 416: 131803.
[104]	MOON M W, PARK S Y, CHOI S K, et al. The phosphotransferase system of Corynebacterium glutamicum: features of sugar transport and carbon regulation[J]. Journal of Molecular Microbiology and Biotechnology, 2007, 12(1-2): 43-50.
[105]	刘冬冬. 强化葡萄糖代谢途径提高L-赖氨酸发酵水平的研究[D]. 无锡: 江南大学, 2017.
	LIU D D. Increasing the fermentation level of L-lysine via enhancing glucose metabolism pathways[D]. Wuxi: Jiangnan University, 2017.
[106]	PARCHE S, BURKOVSKI A, SPRENGER G A, et al. Corynebacterium glutamicum: a dissection of the PTS[J]. Journal of Molecular Microbiology and Biotechnology, 2001, 3(3): 423-428.
[107]	WANG L, LI N, YU S Q, et al. Enhancing caffeic acid production in Escherichia coli by engineering the biosynthesis pathway and transporter[J]. Bioresource Technology, 2023, 368: 128320.
[108]	郭亮, 高聪, 柳亚迪, 等. 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.
	GUO L, GAO C, LIU Y D, et al. Advances in bioproduction of feed amino acid by Escherichia coli [J]. Synthetic Biology Journal, 2021, 2(6): 964-981.
[109]	于勇, 朱欣娜, 毕昌昊, 等. 大肠杆菌细胞工厂的创建技术[J]. 生物工程学报, 2021, 37(5): 1564-1577.
	YU Y, ZHU X N, BI C H, et al. Construction of Escherichia coli cell factories[J]. Chinese Journal of Biotechnology, 2021, 37(5): 1564-1577.
[110]	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 progress on nutrition of several main feed amino acids[J]. China Feed Additive, 2016(1): 10-13.
[111]	ISOGAI S, TAKAGI H. Enhancement of lysine biosynthesis confers high-temperature stress tolerance to Escherichia coli cells[J]. Applied Microbiology and Biotechnology, 2021, 105(18): 6899-6908.
[112]	LIU J, ZHAO X J, CHENG H J, et al. Comprehensive screening of industrially relevant components at genome scale using a high-quality gene overexpression collection of Corynebacterium glutamicum [J]. Trends in Biotechnology, 2025, 43(1): 220-247.
[113]	ZHANG Y W, YANG J J, QIAN F H, et al. Engineering a xylose fermenting yeast for lignocellulosic ethanol production[J]. Nature Chemical Biology, 2025, 21(3): 443-450.
[114]	GAO J Q, LI Y X, YU W, et al. Rescuing yeast from cell death enables overproduction of fatty acids from sole methanol[J]. Nature Metabolism, 2022, 4(7): 932-943.
[115]	GE C, YU Z, SHENG H K, et al. Redesigning regulatory components of quorum-sensing system for diverse metabolic control[J]. Nature Communications, 2022, 13: 2182.
[24]	ZHAO G H, ZHANG D Z, ZHOU B Z, et al. Fine-regulating the carbon flux of L-isoleucine producing Corynebacterium glutamicum WM001 for efficient L-threonine production[J]. ACS Synthetic Biology, 2024, 13(10): 3446-3460.
[25]	PU W, FENG J H, CHEN J Z, et al. Engineering of L-threonine and L-proline biosensors by directed evolution of transcriptional regulator SerR and application for high-throughput screening[J]. Bioresources and Bioprocessing, 2025, 12(1): 4.
[26]	SONG J, ZHUANG M M, DU C Y, et al. Metabolic engineering of Escherichia coli for self-induced production of L-isoleucine[J]. ACS Synthetic Biology, 2025, 14(1): 179-192.
[27]	SHI C R, HUO X J, YOU R, et al. High yield production of L-isoleucine through readjusting the ratio of two direct precursors in Escherichia coli [J]. Bioresource Technology, 2025, 418: 131889.
[28]	LU N, WEI M H, YANG X J, et al. Growth-coupled production of L-isoleucine in Escherichia coli via metabolic engineering[J]. Metabolic Engineering, 2024, 86: 181-193.
[29]	LI Y J, WEI H B, 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(Pt B): 1588-1602.
[30]	LIU Z Y, LIU J, ZHANG F, et al. Modifying Corynebacterium glutamicum by metabolic engineering for efficient synthesis of L-lysine[J]. Systems Microbiology and Biomanufacturing, 2025, 5(1): 288-299.

产品	菌种
液体L-赖氨酸碱	大肠杆菌KCCM 80190	大肠杆菌NITE BP-02917
	谷氨酸棒状杆菌KCCM 80216	谷氨酸棒状杆菌KCTC 12307BP
	谷氨酸棒状杆菌NRRL-B-67439	谷氨酸棒状杆菌NRRL-B-67535
液体L-赖氨酸单盐酸盐	大肠杆菌NITE BP-02917
技术纯L-赖氨酸单盐酸盐	大肠杆菌NITE BP-02917	谷氨酸棒状杆菌NRRL-B-67439
	谷氨酸棒状杆菌DSM 32932	谷氨酸棒状杆菌CGMCC 7.266
	谷氨酸棒状杆菌KCCM 80183	谷氨酸棒状杆菌CGMCC 17927
	谷氨酸棒状杆菌NRRL B-67535	谷氨酸棒状杆菌CCTCC M 2015595
L-赖氨酸硫酸盐	大肠杆菌CGMCC 7.398	谷氨酸棒状杆菌CGMCC 7.266
	谷氨酸棒状杆菌KFCC 11043	谷氨酸棒状杆菌CGMCC 17927
	谷氨酸棒状杆菌KCCM 80227	谷氨酸棒状杆菌CCTCC M 2015595
L-苏氨酸	大肠杆菌DSM 25085	大肠杆菌FERM BP-11383
	大肠杆菌DSM 25086	大肠杆菌FERM BP-10942
	大肠杆菌CGMCC 3703	大肠杆菌NRRL B-30843
	大肠杆菌CGMCC 7.58	大肠杆菌KCCM 11133P
	大肠杆菌CGMCC 7.232	谷氨酸棒状杆菌KCCM 80117
	大肠杆菌CGMCC 13325	谷氨酸棒状杆菌KCCM 80118
L-甲硫氨酸	大肠杆菌KCCM 80245	谷氨酸棒状杆菌KCCM 80246
L-异亮氨酸	大肠杆菌FERM ABP-1064	谷氨酸棒杆菌KCCM 80189
L-异亮氨酸	谷氨酸棒杆菌KCCM 80185	谷氨酸棒杆菌CGMCC 20437

产品	菌种
液体L-赖氨酸碱	大肠杆菌KCCM 80190	大肠杆菌NITE BP-02917
	谷氨酸棒状杆菌KCCM 80216	谷氨酸棒状杆菌KCTC 12307BP
	谷氨酸棒状杆菌NRRL-B-67439	谷氨酸棒状杆菌NRRL-B-67535
液体L-赖氨酸单盐酸盐	大肠杆菌NITE BP-02917
技术纯L-赖氨酸单盐酸盐	大肠杆菌NITE BP-02917	谷氨酸棒状杆菌NRRL-B-67439
	谷氨酸棒状杆菌DSM 32932	谷氨酸棒状杆菌CGMCC 7.266
	谷氨酸棒状杆菌KCCM 80183	谷氨酸棒状杆菌CGMCC 17927
	谷氨酸棒状杆菌NRRL B-67535	谷氨酸棒状杆菌CCTCC M 2015595
L-赖氨酸硫酸盐	大肠杆菌CGMCC 7.398	谷氨酸棒状杆菌CGMCC 7.266
	谷氨酸棒状杆菌KFCC 11043	谷氨酸棒状杆菌CGMCC 17927
	谷氨酸棒状杆菌KCCM 80227	谷氨酸棒状杆菌CCTCC M 2015595
L-苏氨酸	大肠杆菌DSM 25085	大肠杆菌FERM BP-11383
	大肠杆菌DSM 25086	大肠杆菌FERM BP-10942
	大肠杆菌CGMCC 3703	大肠杆菌NRRL B-30843
	大肠杆菌CGMCC 7.58	大肠杆菌KCCM 11133P
	大肠杆菌CGMCC 7.232	谷氨酸棒状杆菌KCCM 80117
	大肠杆菌CGMCC 13325	谷氨酸棒状杆菌KCCM 80118
L-甲硫氨酸	大肠杆菌KCCM 80245	谷氨酸棒状杆菌KCCM 80246
L-异亮氨酸	大肠杆菌FERM ABP-1064	谷氨酸棒杆菌KCCM 80189
L-异亮氨酸	谷氨酸棒杆菌KCCM 80185	谷氨酸棒杆菌CGMCC 20437

酶/前导肽/阻遏蛋白	基因	菌株	策略	效果	参考文献
AK	lysC	E. coli	T344M	L-赖氨酸产量6.3 g/L	[46]
		E. coli	T352I	L-苏氨酸产量14.4 g/L，提升30.9%	[22]
		C. glutamicum	T311I	L-苏氨酸产量0.27 g/L	[47]
		C. glutamicum	S301F	L-苏氨酸产量14.17 g/L，提升4.2%	[24]
	thrA	E. coli	S345F	L-苏氨酸产量82.4 mmol/L	[48]
	thrA	E. coli	G433R	L-苏氨酸产量36.61 mg/L	[49]
AHAS	ilvH	E. coli	G14D, S17F	L-异亮氨酸产量0.322 g/L	[50]
	ilvN	E. coli	H47L	L-异亮氨酸产量5.1 g/L，提升59.4%	[28]
	ilvB	E. coli	K30Q, N156D, V233I	L-异亮氨酸产量5.1 g/L，提升59.4%	[28]
		C. glutamicum	P176S, D426E, L575W	L-异亮氨酸产量4.17 g/L，提升61.0%	[35]
		C. glutamicum	I47Y	L-异亮氨酸产量22.7 g/L，提升10.2%	[51]
TD	ilvA	E. coli	S97F	L-异亮氨酸产量0.322 g/L	[50]
TD	ilvA	C. glutamicum	G96D	L-异亮氨酸产量12 g/L	[24]
ThrL	thrL	E. coli	敲除thrL基因	L-苏氨酸产量1.63 g/L	[21]
MetJ	metJ	E. coli	敲除metJ基因	L-甲硫氨酸产量提升11%	[38]

酶/前导肽/阻遏蛋白	基因	菌株	策略	效果	参考文献
AK	lysC	E. coli	T344M	L-赖氨酸产量6.3 g/L	[46]
		E. coli	T352I	L-苏氨酸产量14.4 g/L，提升30.9%	[22]
		C. glutamicum	T311I	L-苏氨酸产量0.27 g/L	[47]
		C. glutamicum	S301F	L-苏氨酸产量14.17 g/L，提升4.2%	[24]
	thrA	E. coli	S345F	L-苏氨酸产量82.4 mmol/L	[48]
	thrA	E. coli	G433R	L-苏氨酸产量36.61 mg/L	[49]
AHAS	ilvH	E. coli	G14D, S17F	L-异亮氨酸产量0.322 g/L	[50]
	ilvN	E. coli	H47L	L-异亮氨酸产量5.1 g/L，提升59.4%	[28]
	ilvB	E. coli	K30Q, N156D, V233I	L-异亮氨酸产量5.1 g/L，提升59.4%	[28]
		C. glutamicum	P176S, D426E, L575W	L-异亮氨酸产量4.17 g/L，提升61.0%	[35]
		C. glutamicum	I47Y	L-异亮氨酸产量22.7 g/L，提升10.2%	[51]
TD	ilvA	E. coli	S97F	L-异亮氨酸产量0.322 g/L	[50]
TD	ilvA	C. glutamicum	G96D	L-异亮氨酸产量12 g/L	[24]
ThrL	thrL	E. coli	敲除thrL基因	L-苏氨酸产量1.63 g/L	[21]
MetJ	metJ	E. coli	敲除metJ基因	L-甲硫氨酸产量提升11%	[38]

产品	竞争/降解路径	菌株	策略	参考文献
L-苏氨酸	L-赖氨酸、L-甲硫氨酸	E. coli	敲除lysA和metA基因	[49]
	L-赖氨酸、L-甲硫氨酸、L-异亮氨酸、L-甘氨酸	E. coli	敲除lysA、metA和tdh基因，引入ilvA^S97F基因	[22]
	L-赖氨酸、L-异亮氨酸	C. glutamicum	异源表达Streptococcus pneumoniae来源的dapA基因和E. coli来源的ilvA基因	[47]
	L-赖氨酸、L-异亮氨酸、L-丙氨酸	C. glutamicum	引入ilvA^G96D和dapA^K68H基因，敲除alaT和avtA基因	[24]