代谢工程改造微生物合成生物基单体的进展与挑战

doi:10.16085/j.issn.1000-6613.2023-0289

摘要/Abstract

摘要：

单体是合成聚合物所用的小分子基础原料，目前主要来源于化石燃料。利用微生物制备生物基单体具有生产条件温和、环境友好、可持续的优势，是实现高分子材料行业绿色制造的重要途径。借助代谢工程和合成生物学元件，目前已经实现了多种单体的微生物制造，然而与石油基生产工艺相比，这些单体微生物细胞工厂的生产性能普遍较低。围绕代谢工程改造微生物合成生物基单体过程中存在的瓶颈问题，本文基于具体案例分析，从廉价底物的高效利用、提高生物基单体合成效率、强化细胞环境耐受性三个方面，总结了改造微生物合成单体的最新研究进展。同时，讨论了单体微生物细胞工厂目前存在的挑战和未来发展方向。

关键词: 微生物细胞工厂, 塑料单体, 底物利用, 调控策略, 环境耐受性

Abstract:

Monomers are the basic raw materials used in the synthesis of polymers, which mainly come from fossil fuels. Engineering microorganisms to synthesize monomers has the advantages of mild production conditions, environmental friendliness, and sustainability, which is an important way to achieve green manufacturing in the material industry. With the help of metabolic engineering and synthetic biology parts, microbial manufacturing of various monomers has been realized at present. However, compared with petroleum-based production processes, the production performance of these microbial cell factories is limited. Focusing on the bottleneck problems in engineering microorganisms to synthesize bioplastic monomers, this review summarizes the latest research progress in the metabolic engineering of microorganisms to produce monomers from three aspects: efficient utilization of cheap substrates, improvement of monomer synthesis efficiency, and enhancement of cell environment tolerance, based on specific case studies. At the same time, the current challenges and future direction of the microbial monomer cell factory are discussed.

Key words: microbial cell factories, bioplastic monomer, substrate utilization, regulation strategy, environmental tolerance

中图分类号:

Q815

高聪, 陈城虎, 陈修来, 刘立明. 代谢工程改造微生物合成生物基单体的进展与挑战[J]. 化工进展, 2023, 42(8): 4123-4135.

GAO Cong, CHEN Chenghu, CHEN Xiulai, LIU Liming. Progress and challenges of engineering microorganisms to produce biobased monomers[J]. Chemical Industry and Engineering Progress, 2023, 42(8): 4123-4135.

图/表 5

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单体	市场/CNY	菌株	关键调控策略	生产工艺	产量/g·L^-1	参考文献
1,4-BDO	126亿	大肠杆菌	代谢网络模型模拟、合成途径基因组整合	生物发酵	125.0	[81]
		大肠杆菌	全细胞催化赤藓糖醇合成	全细胞转化	4.0×10^-2	[82]
		大肠杆菌	基因编辑调控碳流分布、抑制副产物合成	生物发酵	1.8	[83]
		大肠杆菌	构建谷氨酸转化途径	生物发酵	1.4	[84]
戊二胺	2.20亿	大肠杆菌	蛋白质定向进化	全细胞转化	418	[36]
		大肠杆菌	原位CO₂捕集技术	全细胞转化	209.3	[56]
		大肠杆菌	结合定向进化和计算指导的虚拟筛选	全细胞转化	160.7	[37]
		谷棒杆菌	基因组整合、赖氨酸合成强化	生物发酵	125	[85]
		大肠杆菌	适应性进化	生物发酵	58.7	[86]
		大肠杆菌	温度开关设计、丁二酸联产实现碳固定	生物发酵	55.6	[87]
		大肠杆菌	构建抑制基因表达的sRNAs文库	生物发酵	13.7	[49]
己二酸	60亿	大肠杆菌	平衡路径酶表达水平、阻断副产物积累	生物发酵	68	[88]
		台湾假单胞菌	酶挖掘、级联反应设计、代谢优化	全细胞转化	10.2	[7]
		大肠杆菌	化学溶剂处理和物理破碎方法	生物发酵	55.5	[16]
		酿酒酵母	三阶段发酵工艺优化，筛选异源酶	生物发酵	2.6×10^-3	[31]
		大肠杆菌	建立氧依赖型动态调节系统	生物发酵	0.5	[89]
		酿酒酵母	过表达特异性多药耐药转运蛋白	生物发酵	0.4	[75]
1,3-PDO	7.76亿	大肠杆菌	阻断甘油氧化途径、引入异源途径、失活PTS	生物发酵	135	[90]
		谷棒杆菌	还原力平衡、降低有毒中间代谢物积累	生物发酵	110	[91]
		罗伊氏乳杆菌	电子束辐照诱变	生物发酵	93.2	[74]
		肺炎克雷伯菌	敲除葡萄糖转运蛋白Crr	生物发酵	78	[26]
		需钠弧菌	删除全局转录调节因子，转录组学分析	生物发酵	56.2	[51]
		丁酸梭菌	组合NTG和ARTP诱变筛选	生物发酵	37	[73]
		大肠杆菌	微调混合途径以匹配合成路径的能源需求	生物发酵	22.7	[92]
乳酸	29亿	大肠杆菌	阻断副产物合成	生物发酵	142.2	[93]
		酿酒酵母	引入耐受蛋白IoGAS1	生物发酵	92	[63]
		大肠杆菌	筛选木糖分解代谢操纵子	生物发酵	86	[25]
		恶臭假单胞菌、凝结芽孢杆菌	合成微生物群落	生物发酵	35.8	[20]
		食木薯乳杆菌	聚集用于底物的同时糖化和发酵	生物发酵	18.7	[28]
		里氏木霉、戴尔根霉	合成微生物群落	生物发酵	4.4	[11]
对苯二甲酸	495亿	大肠杆菌	基因组多拷贝整合	全细胞转化	38.3	[15]
对苯二甲酸	495亿	大肠杆菌	微生物双相转化	全细胞转化	6.9	[14]
丁二酸	2.23亿	产琥珀酸曼氏杆菌	筛选不同来源的苹果酸脱氢酶	生物发酵	134.3	[30]
		大肠杆菌	溶氧动态开关	生物发酵	119	[40]
		产琥珀酸曼氏杆菌	改变细胞膜的流动性	生物发酵	84.2	[59]
		产琥珀酸放线杆菌	生物被膜的细胞固定化	生物发酵	67.3	[94]
		大肠杆菌	替代高能耗途径	生物发酵	56.9	[52]
		大肠杆菌	过表达大肠杆菌全局转录调控因子IrrE	生物发酵	24.5	[68]
		产丙酸丙酸杆菌	原位产物分离，开发膜分离耦合发酵技术	生物发酵	20.5	[76]
戊二酸	4100万	谷棒杆菌	基因组分析、组合优化工程	生物发酵	105.3	[48]
		大肠杆菌	质粒优化、启动子工程和核糖体结合位点工程	全细胞转化	77.6	[32]
		大肠杆菌	胶体几丁质细胞固定策略	全细胞转化	73.2	[38]
		大肠杆菌	表达戊二胺转运蛋白、耦合辅因子驱动技术	生物发酵	54.5	[34]
		大肠杆菌	针对副产物合成途径的自主回路	生物发酵	26	[42]
		谷棒杆菌	适应性进化	生物发酵	22.7	[3]
		大肠杆菌	引入丙二酸利用途径	生物发酵	6.3	[95]

单体	市场/CNY	菌株	关键调控策略	生产工艺	产量/g·L^-1	参考文献
1,4-BDO	126亿	大肠杆菌	代谢网络模型模拟、合成途径基因组整合	生物发酵	125.0	[81]
		大肠杆菌	全细胞催化赤藓糖醇合成	全细胞转化	4.0×10^-2	[82]
		大肠杆菌	基因编辑调控碳流分布、抑制副产物合成	生物发酵	1.8	[83]
		大肠杆菌	构建谷氨酸转化途径	生物发酵	1.4	[84]
戊二胺	2.20亿	大肠杆菌	蛋白质定向进化	全细胞转化	418	[36]
		大肠杆菌	原位CO₂捕集技术	全细胞转化	209.3	[56]
		大肠杆菌	结合定向进化和计算指导的虚拟筛选	全细胞转化	160.7	[37]
		谷棒杆菌	基因组整合、赖氨酸合成强化	生物发酵	125	[85]
		大肠杆菌	适应性进化	生物发酵	58.7	[86]
		大肠杆菌	温度开关设计、丁二酸联产实现碳固定	生物发酵	55.6	[87]
		大肠杆菌	构建抑制基因表达的sRNAs文库	生物发酵	13.7	[49]
己二酸	60亿	大肠杆菌	平衡路径酶表达水平、阻断副产物积累	生物发酵	68	[88]
		台湾假单胞菌	酶挖掘、级联反应设计、代谢优化	全细胞转化	10.2	[7]
		大肠杆菌	化学溶剂处理和物理破碎方法	生物发酵	55.5	[16]
		酿酒酵母	三阶段发酵工艺优化，筛选异源酶	生物发酵	2.6×10^-3	[31]
		大肠杆菌	建立氧依赖型动态调节系统	生物发酵	0.5	[89]
		酿酒酵母	过表达特异性多药耐药转运蛋白	生物发酵	0.4	[75]
1,3-PDO	7.76亿	大肠杆菌	阻断甘油氧化途径、引入异源途径、失活PTS	生物发酵	135	[90]
		谷棒杆菌	还原力平衡、降低有毒中间代谢物积累	生物发酵	110	[91]
		罗伊氏乳杆菌	电子束辐照诱变	生物发酵	93.2	[74]
		肺炎克雷伯菌	敲除葡萄糖转运蛋白Crr	生物发酵	78	[26]
		需钠弧菌	删除全局转录调节因子，转录组学分析	生物发酵	56.2	[51]
		丁酸梭菌	组合NTG和ARTP诱变筛选	生物发酵	37	[73]
		大肠杆菌	微调混合途径以匹配合成路径的能源需求	生物发酵	22.7	[92]
乳酸	29亿	大肠杆菌	阻断副产物合成	生物发酵	142.2	[93]
		酿酒酵母	引入耐受蛋白IoGAS1	生物发酵	92	[63]
		大肠杆菌	筛选木糖分解代谢操纵子	生物发酵	86	[25]
		恶臭假单胞菌、凝结芽孢杆菌	合成微生物群落	生物发酵	35.8	[20]
		食木薯乳杆菌	聚集用于底物的同时糖化和发酵	生物发酵	18.7	[28]
		里氏木霉、戴尔根霉	合成微生物群落	生物发酵	4.4	[11]
对苯二甲酸	495亿	大肠杆菌	基因组多拷贝整合	全细胞转化	38.3	[15]
对苯二甲酸	495亿	大肠杆菌	微生物双相转化	全细胞转化	6.9	[14]
丁二酸	2.23亿	产琥珀酸曼氏杆菌	筛选不同来源的苹果酸脱氢酶	生物发酵	134.3	[30]
		大肠杆菌	溶氧动态开关	生物发酵	119	[40]
		产琥珀酸曼氏杆菌	改变细胞膜的流动性	生物发酵	84.2	[59]
		产琥珀酸放线杆菌	生物被膜的细胞固定化	生物发酵	67.3	[94]
		大肠杆菌	替代高能耗途径	生物发酵	56.9	[52]
		大肠杆菌	过表达大肠杆菌全局转录调控因子IrrE	生物发酵	24.5	[68]
		产丙酸丙酸杆菌	原位产物分离，开发膜分离耦合发酵技术	生物发酵	20.5	[76]
戊二酸	4100万	谷棒杆菌	基因组分析、组合优化工程	生物发酵	105.3	[48]
		大肠杆菌	质粒优化、启动子工程和核糖体结合位点工程	全细胞转化	77.6	[32]
		大肠杆菌	胶体几丁质细胞固定策略	全细胞转化	73.2	[38]
		大肠杆菌	表达戊二胺转运蛋白、耦合辅因子驱动技术	生物发酵	54.5	[34]
		大肠杆菌	针对副产物合成途径的自主回路	生物发酵	26	[42]
		谷棒杆菌	适应性进化	生物发酵	22.7	[3]
		大肠杆菌	引入丙二酸利用途径	生物发酵	6.3	[95]

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