等离子体微生物育种技术在生物制造中的应用进展

doi:10.12211/2096-8280.2025-005

摘要/Abstract

摘要：

可持续绿色生物制造是各国关注的战略重点，但微生物性能仍是制约生物制造产业化的瓶颈之一。传统育种方法常面临育种周期长、效率低、成本高等问题，难以满足工业化生产对高效、稳定生产菌株的需求。低温等离子体技术作为一种高效、绿色环保的微生物育种方法，可通过刺激突变位点、提升突变效率、扩展突变范围，有效提高目标菌株性能和产品产量，为工业微生物改良提供重要助力。本文综述了等离子体诱变技术的理论基础、三种等离子体源（ARTP、DBD、CD）的技术特点、等离子体诱变作用机制及其与高通量筛选、传统诱变、理性育种等技术的联用进展，归纳出生物酶、有机酸、生物能源、生物材料等生物制造领域的典型等离子体育种案例，为相关领域的研究和产业化提供参考。未来需要开发基于空气源的新型等离子体发生器，在小型化的基础上实现低成本、低能耗、低温升；与高通量筛选和AI等技术相融合进行菌株精准诱变和高效育种，实现技术瓶颈突破，最终推动生物制造产业升级。

关键词: 生物制造, 工业微生物, 等离子体诱变技术, 筛选, 诱变机制, 生物材料

Abstract:

Sustainable and green biomanufacturing has emerged as a strategic priority for nations globally; however, microbial performance remains one of the key constraints hindering the industrialization of biomanufacturing. Traditional breeding methods often face challenges such as long breeding cycles, low efficiency, and high costs, making it difficult to meet the demands for highly efficient and stable microbial strains in industrial-scale production. Low-temperature plasma technology, as an efficient and environmentally friendly method for microbial breeding, can stimulate mutation hotspots, enhance mutation efficiency, and expand mutation ranges, effectively improving the performance of target strains and product yields. By combining plasma mutagenesis’s advanced capabilities with other techniques, significant improvements in the performance and productivity of microbial strains can be achieved, thus driving the commercialization of sustainable bioprocesses. This review outlines the theoretical basis of plasma mutagenesis technology, the technical characteristics of three plasma sources (ARTP, DBD, and CD), the mechanisms of plasma mutagenesis, and the progress of combining this technology with high-throughput screening, classic mutagenesis, and rational breeding methods. Furthermore, it summarizes typical plasma breeding cases in biomanufacturing fields such as bio-enzyme, organic acids, bioenergy, and biomaterials. These insights offer important references for research and industrialization in related fields. By combining plasma mutagenesis’s advanced capabilities with other techniques, significant improvements in the performance and productivity of microbial strains can be achieved, thus driving the commercialization of sustainable bioprocesses. The cases discussed in this review provide a practical understanding of how plasma mutagenesis can be applied to optimize microbial strains for industrial-scale production of valuable bioproducts, providing a reference for research and industrialization in related fields. In the future, it is essential to develop novel plasma generators based on air source, which, while being miniaturized, achieve low cost, low energy consumption, and minimal temperature rise. Integrating them with high-throughput screening and AI technologies will enable precise microbial mutagenesis and efficient strain breeding, thereby overcoming technical bottlenecks and ultimately advancing the biomanufacturing industry. {L-End}

Key words: biological manufacturing, industrial microbiology, plasma mutagenesis technology, screening, mutagenesis mechanism, biological materials

中图分类号:

Q819

钟奶才, 陈缘, 潘文锋, 苏小凤, 廖景文, 翟英雷, 钟近艺. 等离子体微生物育种技术在生物制造中的应用进展[J]. 合成生物学, 2025, 6(4): 789-805.

ZHONG Naicai, CHEN Yuan, PAN Wenfeng, SU Xiaofeng, LIAO Jingwen, ZHAI Yinglei, ZHONG Jinyi. Application of plasma microbial breeding technology in biofabrication[J]. Synthetic Biology Journal, 2025, 6(4): 789-805.

图/表 8

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等离子体联用技术	优点	缺点
高通量筛选技术	筛选效率高，筛选范围广通量高	设备成本要求高，操作复杂
传统诱变方法	互补性强突变位点广，突破单一方法局限	操作烦琐，遗传稳定性差
理性突变方法	精准性高，突变效果明显	技术要求高，成本较高，伦理限制

等离子体联用技术	优点	缺点
高通量筛选技术	筛选效率高，筛选范围广通量高	设备成本要求高，操作复杂
传统诱变方法	互补性强突变位点广，突破单一方法局限	操作烦琐，遗传稳定性差
理性突变方法	精准性高，突变效果明显	技术要求高，成本较高，伦理限制

应用领域	诱变菌株	诱变目的	评价参数	实验结果	参考文献
酶	红平菇	漆酶	酶活	酶活提高86.36%，遗传稳定	[64]
	莫巴拉链霉菌	谷氨酰胺转氨酶	比活性	TGase提高35.6倍和2.9倍	[65]
	米曲霉	蛋白酶	酶活	酶活及相关关键基因表达提高	[66]
	黑曲霉	单宁酶	酶活	酶活提高2.27倍，具良好遗传稳定性	[67]
	高山被孢霉	花生四烯酸（ARA）	花生四烯酸产量	ARA总脂肪酸占比及总产量分别提高75.40%和232.21%	[68]
脂肪酸	大肠杆菌	L-半胱氨酸	滴度	L-半胱氨酸滴度增加了2.2倍	[69]
	谷氨酸棒状杆菌	谷氨酸	耐高糖和丙二酸	合成关键基因表达提高	[70]
	酿酒酵母	酪氨酸	前体香豆酸p-CA滴度	p-CA产量提高7.6倍	[71]
	裂殖壶菌	二十二碳六烯酸（DHA）	关键酶表达及DHA产量	DHA产量提高14.3%	[72]
	琥珀酸放线杆菌	琥珀酸	抗逆性、产量	琥珀酸环境抗逆性提高2倍，产量增加113%	[73]
生物药物	马杜拉放线菌	喷司他丁	喷司他丁产量	喷司他丁产量增加33.79%	[74]
	短孢杆菌SPR20	抗菌肽	MIC值	MIC值达到250～500 μg/mL和500 μg/mL	[75]
	玫瑰链霉菌	达托霉素	达托霉素产量	达托霉素产量提高58.33%	[76]
	大肠杆菌E. coli NXBG-13	胞苷	胞苷滴度、产量、生产率	胞苷的效价、产量和生产率达到15.7 g/L, 0.164 g/g和0.327 g/(L·h)	[60]
	桑黄多孔菌	胞内多糖	多糖产量	多糖产量提高1.5倍	[77]
生物燃料	丁酸梭菌	1,3-丙二醇 (1,3-PD)	1,3-PD滴度	1,3-PD滴度提高4.88倍	[78]
	嗜热毁丝霉	木聚糖酶	酶活、耐酸性	木聚糖酶活性提高21.71%，酸性（pH 4.0～7.0）适应性提高	[79]
	酿酒酵母	乙醇	乙醇及前体产量	乙酸乙酯和乙酸异戊酯浓度提高2.8倍和3.3倍	[80]
	卷枝毛霉	生物柴油	甲醇耐受性、生物柴油产率	甲醇耐受性提高，柴油产率达到91%	[81]
	热带念珠菌	脂质	脂质产量	脂质产量增强16.8%	[82]
	莱茵衣藻	氢气	氢气产量	氢气产量增加1.8～5.2倍和2.7～3.1倍	[83]
	凯斯小球藻	生物柴油、脂质	生物柴油、脂质产量	生物量和脂质生产力提高75%和44%	[84]
生物材料	嗜盐单胞菌	聚羟基脂肪酸酯	盐胁迫条件	生产盐度降低，PHA生产成本降低33%	[85]
	出芽短梗霉	普鲁兰多糖	普鲁兰多糖产量	普鲁兰多糖产量提高17.6%	[86]
	蜡样芽孢杆菌	壳聚糖酶	酶活	壳聚糖酶活性提高2.49倍	[87]
	紫红曲霉	色素	橙色素、橘霉素含量	橙色素产量提高66.7%，橘霉素降低69%	[88]
	克雷伯氏菌	苯二甲酸丙二醇酯单体1,3-丙二醇	1,3-丙二醇产量，甘油转化率	1,3-丙二醇产量达到118 g/L，甘油转化率为42%	[89]

应用领域	诱变菌株	诱变目的	评价参数	实验结果	参考文献
酶	红平菇	漆酶	酶活	酶活提高86.36%，遗传稳定	[64]
	莫巴拉链霉菌	谷氨酰胺转氨酶	比活性	TGase提高35.6倍和2.9倍	[65]
	米曲霉	蛋白酶	酶活	酶活及相关关键基因表达提高	[66]
	黑曲霉	单宁酶	酶活	酶活提高2.27倍，具良好遗传稳定性	[67]
	高山被孢霉	花生四烯酸（ARA）	花生四烯酸产量	ARA总脂肪酸占比及总产量分别提高75.40%和232.21%	[68]
脂肪酸	大肠杆菌	L-半胱氨酸	滴度	L-半胱氨酸滴度增加了2.2倍	[69]
	谷氨酸棒状杆菌	谷氨酸	耐高糖和丙二酸	合成关键基因表达提高	[70]
	酿酒酵母	酪氨酸	前体香豆酸p-CA滴度	p-CA产量提高7.6倍	[71]
	裂殖壶菌	二十二碳六烯酸（DHA）	关键酶表达及DHA产量	DHA产量提高14.3%	[72]
	琥珀酸放线杆菌	琥珀酸	抗逆性、产量	琥珀酸环境抗逆性提高2倍，产量增加113%	[73]
生物药物	马杜拉放线菌	喷司他丁	喷司他丁产量	喷司他丁产量增加33.79%	[74]
	短孢杆菌SPR20	抗菌肽	MIC值	MIC值达到250～500 μg/mL和500 μg/mL	[75]
	玫瑰链霉菌	达托霉素	达托霉素产量	达托霉素产量提高58.33%	[76]
	大肠杆菌E. coli NXBG-13	胞苷	胞苷滴度、产量、生产率	胞苷的效价、产量和生产率达到15.7 g/L, 0.164 g/g和0.327 g/(L·h)	[60]
	桑黄多孔菌	胞内多糖	多糖产量	多糖产量提高1.5倍	[77]
生物燃料	丁酸梭菌	1,3-丙二醇 (1,3-PD)	1,3-PD滴度	1,3-PD滴度提高4.88倍	[78]
	嗜热毁丝霉	木聚糖酶	酶活、耐酸性	木聚糖酶活性提高21.71%，酸性（pH 4.0～7.0）适应性提高	[79]
	酿酒酵母	乙醇	乙醇及前体产量	乙酸乙酯和乙酸异戊酯浓度提高2.8倍和3.3倍	[80]
	卷枝毛霉	生物柴油	甲醇耐受性、生物柴油产率	甲醇耐受性提高，柴油产率达到91%	[81]
	热带念珠菌	脂质	脂质产量	脂质产量增强16.8%	[82]
	莱茵衣藻	氢气	氢气产量	氢气产量增加1.8～5.2倍和2.7～3.1倍	[83]
	凯斯小球藻	生物柴油、脂质	生物柴油、脂质产量	生物量和脂质生产力提高75%和44%	[84]
生物材料	嗜盐单胞菌	聚羟基脂肪酸酯	盐胁迫条件	生产盐度降低，PHA生产成本降低33%	[85]
	出芽短梗霉	普鲁兰多糖	普鲁兰多糖产量	普鲁兰多糖产量提高17.6%	[86]
	蜡样芽孢杆菌	壳聚糖酶	酶活	壳聚糖酶活性提高2.49倍	[87]
	紫红曲霉	色素	橙色素、橘霉素含量	橙色素产量提高66.7%，橘霉素降低69%	[88]
	克雷伯氏菌	苯二甲酸丙二醇酯单体1,3-丙二醇	1,3-丙二醇产量，甘油转化率	1,3-丙二醇产量达到118 g/L，甘油转化率为42%	[89]