5-氨基乙酰丙酸（5-ALA）的生物合成研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2025-0153

摘要/Abstract

摘要：

5-氨基乙酰丙酸（5-ALA）是生物体内天然存在一种非蛋白质类氨基酸，目前已被广泛应用于医药、保健、农业和动物添加剂等领域。5-ALA主要通过化学和生物法合成，但由于化学合成的工艺复杂性和成本问题限制了其规模化发展，而生物合成法以其环保、高效等优势彰显了其工业化发展的巨大潜力。近年来，由于合成生物学和代谢工程等交叉学科的迅猛发展，5-ALA的生物合成已逐渐成为研究热点。本文回溯了过去几十年5-ALA生物合成的发展历程，总结并梳理了基于C₄和C₅途径、辅因子再生途径、三羧酸循环等途径、用于高效细胞工厂构建的各种代谢工程策略的研究进展，重点阐述了基于生物传感器的动态调控和高通量筛选方法的建立等策略在菌株代谢工程改造中的应用和重要性，并就进一步提高5-ALA的合成产量，打破其工业化应用的瓶颈提出了几点策略和展望，以期为5-ALA的高效合成和产业制造提供一定的参考和研究依据。

关键词: 5-氨基乙酰丙酸, 生物合成, 代谢工程, 动态调控, 生物传感器

Abstract:

5-aminolevulinic acid (5-ALA) is a non-protein amino acid naturally occurring in living organisms, and has been widely used in the fields of medicine, health care, agriculture and animal additives, etc. 5-ALA is mainly synthesized by chemical and biological methods, but the complexity of the chemical synthesis process and cost problems have limited its large-scale development, and biosynthesis method has highlighted its great potential for industrial development due to its advantages of environmental protection and high efficiency. The biosynthesis method, with its advantages of environmental protection and high efficiency, has shown its great potential for industrial development. In recent years, the biosynthesis of 5-ALA has gradually become a research hotspot due to the rapid development of interdisciplinary disciplines such as synthetic biology and metabolic engineering. In this paper, we reviewed the development of 5-ALA biosynthesis in the past decades, summarized the progress of various metabolic engineering strategies based on the C₄ and C₅ pathways, cofactor regeneration pathway, tricarboxylic acid cycle and other pathways for the construction of high-efficiency cell factories, and highlighted the application and significance of the dynamic regulation based on biosensors and the establishment of high-throughput screening methods in the metabolic engineering modification of the bacterial strains. We also put forward several strategies and prospects for further improving the synthetic yield of 5-ALA and breaking the bottleneck of its industrial application, with a view to providing certain references and research bases for the efficient synthesis and industrial manufacturing of 5-ALA.

Key words: 5-aminolevulinic acid, biosynthesis, metabolic engineering, dynamic regulation, biosensors

吕欢欢, 张高阳, 王赛笛, 孙忠科, 李成伟, 罗德平. 5-氨基乙酰丙酸（5-ALA）的生物合成研究进展[J]. 生物技术通报, 2025, 41(11): 75-88.

LYU Huan-huan, ZHANG Gao-yang, WANG Sai-di, SUN Zhong-ke, LI Cheng-wei, LUO De-ping. Advances in the Biosynthesis of 5-aminolevulinic Acid （5-ALA）[J]. Biotechnology Bulletin, 2025, 41(11): 75-88.

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菌株 Strain	底物 Substrate	策略 Strategy	滴度 Titer （g/L）	参考文献 Reference
大肠杆菌 E. coli	葡萄糖Glucose 甘油Glycerol	过表达荚膜红细菌来源的ALAS酶并进行密码子优化，过表达伴侣蛋白GroELS，培养基中添加铁离子	15.6	［39］
	葡萄糖Glucose 谷氨酸Glutamate	异源表达拟南芥来源的hemA基因和GluTR调节蛋白PGR7，优化宿主菌株	7.64	［40］
	葡萄糖Glucose	选择不同来源的C5途径关键酶并优化其表达比例，微调竞争性代谢途径，优化辅因子再生途径	12.1	［41］
大肠杆菌 E. coliB	葡萄糖Glucose 甘氨酸Glycine	过表达荚膜红细菌来源的ALAS酶，加强PLP补救途径	8.21	［42］
	葡萄糖Glucose 甘氨酸Glycine琥珀酸Succinate	过表达球形红杆菌来源的hemA基因，过表达外排蛋白基因rhtA，过表达pdxK和pdxY基因以增加辅因子PLP的再生，引入伴侣蛋白DnaK和GroELS	7.47	［43］
	葡萄糖Glucose	过表达ALAS酶，过表达过氧化氢酶CAT和超氧化物歧化酶SOD以增强抗氧化系统	11.5	［44］
	葡萄糖Glucose 磷酸吡哆醛PLP 泛酸PA	进化分析筛选新的ALAS酶，识别并修改4个关键靶标，开发动态控制系统以平衡氧化还原稳态和碳通量，协同优化辅因子	63.39	［11］
	葡萄糖Glucose	过表达hemA^RS 基因，基于RGMS技术的eamA基因定向进化，过表达sodB和katE基因提高抗氧化性，减弱hemB基因的表达	19.02	［45］
	葡萄糖Glucose 甘氨酸Glycine	建立5-ALA响应生物传感器，筛选HemA酶和菌株突变体，加强5-ALA外排，抑制下游基因表达，提高前体和辅因子供应	58.54	［46］
谷氨酸棒状杆菌 C. glutamicum	葡萄糖Glucose	过表达荚膜红细菌来源的hemA基因，过表达外排蛋白基因rhtA，敲除sucCD基因，采用“生长-产酸”两阶段发酵模式	14.7	［20］
	葡萄糖Glucose	RBS工程优化RphemA基因的表达，敲除odhI和sdhA基因，用small RNA抑制hemB基因的表达，过表达外排蛋白基因eamA	25.05	［47］
	葡萄糖Glucose	定向诱变odhI基因，增加谷氨酸供应，添加诱导剂吐温40和乙胺丁醇	2.9	［48］
	葡萄糖Glucose 甘氨酸Glycine	过表达球形红杆菌来源的ALAS酶，削弱竞争途径，过表达ppc基因和外排蛋白基因rhtA，敲除细胞壁合成相关蛋白HMW-PBP	7.53	［49］
	葡萄糖Glucose 木薯渣Cassava bagasse	筛选比较不同来源的ALAS酶，RBS工程优化hemA基因和ppc基因平衡表达，利用廉价可再生原料替代葡萄糖	18.5	［50］
	葡萄糖Glucose	通过3种不同策略调节odhA基因表达，响应血红素浓度变化自动调节rhtA基因表达，精确调控5-ALA合成和辅因子再生关键基因	3.16	［51］

菌株 Strain	底物 Substrate	策略 Strategy	滴度 Titer （g/L）	参考文献 Reference
大肠杆菌 E. coli	葡萄糖Glucose 甘油Glycerol	过表达荚膜红细菌来源的ALAS酶并进行密码子优化，过表达伴侣蛋白GroELS，培养基中添加铁离子	15.6	［39］
	葡萄糖Glucose 谷氨酸Glutamate	异源表达拟南芥来源的hemA基因和GluTR调节蛋白PGR7，优化宿主菌株	7.64	［40］
	葡萄糖Glucose	选择不同来源的C5途径关键酶并优化其表达比例，微调竞争性代谢途径，优化辅因子再生途径	12.1	［41］
大肠杆菌 E. coliB	葡萄糖Glucose 甘氨酸Glycine	过表达荚膜红细菌来源的ALAS酶，加强PLP补救途径	8.21	［42］
	葡萄糖Glucose 甘氨酸Glycine琥珀酸Succinate	过表达球形红杆菌来源的hemA基因，过表达外排蛋白基因rhtA，过表达pdxK和pdxY基因以增加辅因子PLP的再生，引入伴侣蛋白DnaK和GroELS	7.47	［43］
	葡萄糖Glucose	过表达ALAS酶，过表达过氧化氢酶CAT和超氧化物歧化酶SOD以增强抗氧化系统	11.5	［44］
	葡萄糖Glucose 磷酸吡哆醛PLP 泛酸PA	进化分析筛选新的ALAS酶，识别并修改4个关键靶标，开发动态控制系统以平衡氧化还原稳态和碳通量，协同优化辅因子	63.39	［11］
	葡萄糖Glucose	过表达hemA^RS 基因，基于RGMS技术的eamA基因定向进化，过表达sodB和katE基因提高抗氧化性，减弱hemB基因的表达	19.02	［45］
	葡萄糖Glucose 甘氨酸Glycine	建立5-ALA响应生物传感器，筛选HemA酶和菌株突变体，加强5-ALA外排，抑制下游基因表达，提高前体和辅因子供应	58.54	［46］
谷氨酸棒状杆菌 C. glutamicum	葡萄糖Glucose	过表达荚膜红细菌来源的hemA基因，过表达外排蛋白基因rhtA，敲除sucCD基因，采用“生长-产酸”两阶段发酵模式	14.7	［20］
	葡萄糖Glucose	RBS工程优化RphemA基因的表达，敲除odhI和sdhA基因，用small RNA抑制hemB基因的表达，过表达外排蛋白基因eamA	25.05	［47］
	葡萄糖Glucose	定向诱变odhI基因，增加谷氨酸供应，添加诱导剂吐温40和乙胺丁醇	2.9	［48］
	葡萄糖Glucose 甘氨酸Glycine	过表达球形红杆菌来源的ALAS酶，削弱竞争途径，过表达ppc基因和外排蛋白基因rhtA，敲除细胞壁合成相关蛋白HMW-PBP	7.53	［49］
	葡萄糖Glucose 木薯渣Cassava bagasse	筛选比较不同来源的ALAS酶，RBS工程优化hemA基因和ppc基因平衡表达，利用廉价可再生原料替代葡萄糖	18.5	［50］
	葡萄糖Glucose	通过3种不同策略调节odhA基因表达，响应血红素浓度变化自动调节rhtA基因表达，精确调控5-ALA合成和辅因子再生关键基因	3.16	［51］