大肠杆菌Nissle 1917的合成生物学平台开发及应用进展

doi:10.13560/j.cnki.biotech.bull.1985.2025-0713

摘要/Abstract

摘要：

作为在多个领域具有广泛应用价值的益生菌，大肠杆菌属Nissle 1917（EcN）凭借其临床验证的安全性、优异的肠道定植能力以及与模式菌株的遗传兼容性，被认为是理想的工程化底盘微生物平台。因此，开发适用于EcN的成熟遗传操作体系，是实现其在不同领域功能化应用的基础。本文系统综述了EcN的益生菌特性，重点总结了其在遗传编辑工具开发方面的最新进展，梳理了通过基因工程手段改造EcN在疾病（如炎症性肠病、肿瘤等）治疗的药物递送系统以及高效生物合成中的应用，并进一步探讨了人工智能等前沿技术的赋能助力下，EcN工程菌在精准安全基因编辑工具开发、活体药物开发与绿色生物制造领域的发展方向与潜力。本文旨在推动EcN平台在不同领域的广泛应用，并为未来的研究与产业化应用提供参考与启示。

关键词: 大肠杆菌Nissle 1917, 益生菌, 调控元件, 疾病治疗, 生物合成

Abstract:

As a probiotic with broad application value across multiple fields, Escherichia coli Nissle 1917 (EcN) is recognized as an ideal engineered chassis microbial platform, owing to its clinically validated safety, superior gut colonization capacity, and genetic compatibility with model strains. Consequently, the development of a mature genetic manipulation system for EcN is fundamental to achieving its functional applications in diverse areas. This article systematically reviews the probiotic properties of EcN, with a focus on summarizing recent advances in the development of genetic editing tools for this strain. It further sorts applications of genetically engineered EcN in therapeutic drug delivery systems for diseases (such as inflammatory bowel disease and tumor) and in efficient biosynthesis. Finally, the review explores the future directions and potential of engineered EcN strains, empowered by cutting-edge technologies like artificial intelligence, in the development of precise and safe gene editing tools, live biotherapeutics, and green biomanufacturing. This work aims to promote the widespread application of the EcN platform across various fields and provide insights and references for future research and industrial translation.

Key words: Escherichia coli Nissle 1917, probiotics, regulatory elements, treatment of disease, synthetic biology

周思延, 丁炜权, 董平, 翁含之, 胥睿睿, 康振. 大肠杆菌Nissle 1917的合成生物学平台开发及应用进展[J]. 生物技术通报, 2025, 41(11): 47-61.

ZHOU Si-yan, DING Wei-quan, DONG Ping, WENG Han-zhi, XU Rui-rui, KANG Zhen. Advances in Synthetic Biology Platform Development and Application for Escherichiacoli Nissle 1917[J]. Biotechnology Bulletin, 2025, 41(11): 47-61.

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工具 Tool	类型 Type	用途 Aplication	文献来源 Reference
启动子	组成型	以J23100启动子表达gabB基因，提高GABA的产量	［41］
	组成型	通过P_fnrS启动子表达phaA、phaB、tesB、mailKE基因，提高3HB的产量	［42］
	化学诱导	T7表达系统表达铁氧还蛋白氧化还原酶PcyA	［47］
		通过T7表达系统表达基因产生血红素代谢物	［44］
		通过T7表达系统同时合成磺基转移酶和肝素前体	［46］
		通过lac启动子表达kfiC基因控制荚膜	［48］
	光诱导	通过蓝光负责启动子表达Ag43、IL-10、TNFα、FlaB，进行精准治疗	［50］
	光诱导	红光遗传电路裂解细胞释放组成表达的Ex-4-aa	［56］
CRISPR/Cas	CRISPRi	抑制ccdAB和hipAB基因表达平衡生物膜的定植和毒性	［61］
	CRISPRi	csgD基因的抑制作用	［60］
	基因编辑	敲除frdA、ldhA、adhE和pta基因，提高GABA的产量	［41］
		敲除内源隐性质粒	［72］
		作为一种靶向抗生素耐药基因的抗菌药物	［62］
“自杀”系统	光诱导	触发毒素蛋白CcdB的积累和抗毒素蛋白CcdA的减少	［52］
	CRISPR	引导Cas9到达切割基因组的目标序列	［66］
	群体感应	触发lysin的表达，释放蛋白S5和DspB	［70］
	群体感应	lux启动子驱动的luxI转录，以及噬菌体衍生的裂解基因φX174E	［67］

工具 Tool	类型 Type	用途 Aplication	文献来源 Reference
启动子	组成型	以J23100启动子表达gabB基因，提高GABA的产量	［41］
	组成型	通过P_fnrS启动子表达phaA、phaB、tesB、mailKE基因，提高3HB的产量	［42］
	化学诱导	T7表达系统表达铁氧还蛋白氧化还原酶PcyA	［47］
		通过T7表达系统表达基因产生血红素代谢物	［44］
		通过T7表达系统同时合成磺基转移酶和肝素前体	［46］
		通过lac启动子表达kfiC基因控制荚膜	［48］
	光诱导	通过蓝光负责启动子表达Ag43、IL-10、TNFα、FlaB，进行精准治疗	［50］
	光诱导	红光遗传电路裂解细胞释放组成表达的Ex-4-aa	［56］
CRISPR/Cas	CRISPRi	抑制ccdAB和hipAB基因表达平衡生物膜的定植和毒性	［61］
	CRISPRi	csgD基因的抑制作用	［60］
	基因编辑	敲除frdA、ldhA、adhE和pta基因，提高GABA的产量	［41］
		敲除内源隐性质粒	［72］
		作为一种靶向抗生素耐药基因的抗菌药物	［62］
“自杀”系统	光诱导	触发毒素蛋白CcdB的积累和抗毒素蛋白CcdA的减少	［52］
	CRISPR	引导Cas9到达切割基因组的目标序列	［66］
	群体感应	触发lysin的表达，释放蛋白S5和DspB	［70］
	群体感应	lux启动子驱动的luxI转录，以及噬菌体衍生的裂解基因φX174E	［67］

疾病 Disease	EcN的作用 Effect of EcN	动物模型/细胞系 Animal model/Cell line	参考文献 References
多重致敏	Th2淋巴细胞减少	鼠	［79］
感染	免疫调节	猪	［96］
过敏性哮喘	抑制T2和ILC2的激活	鼠	［97］
志贺氏菌感染	干扰噬菌体感染易感大肠杆菌	大肠杆菌	［98］
空肠弯曲杆菌感染	调节保护性先天免疫相关基因的表达	HT-29细胞	［99］
非传染性葡萄膜炎	调节肠眼轴	鼠	［100］
肠上皮屏障功能障碍	维持与TJ结构相关的亚细胞定位	T-84和Caco-2细胞	［2］
人轮状病毒感染	增强先天和B细胞免疫反应	猪	［77］
结肠癌	通过调节信号通路诱导细胞凋亡	HT-29细胞	［3］
IBD	调节特定mRNA的表达	鼠	［6］
呼吸道合胞病毒	逆转肠道生态失调	鼠	［101］
抗生素不适	阻断病理肠脑回路	鼠	［102］
慢性细菌性前列腺炎	左氧氟沙星联合治疗	人体	［103］
肠道炎症性/感染性腹泻	外膜囊泡调节免疫细胞的功能	RAW 264.7细胞	［104］
苯丙酮尿症	降解丙氨酸	人体	［89］
口腔癌	检测乳酸	人体	［105］

疾病 Disease	EcN的作用 Effect of EcN	动物模型/细胞系 Animal model/Cell line	参考文献 References
多重致敏	Th2淋巴细胞减少	鼠	［79］
感染	免疫调节	猪	［96］
过敏性哮喘	抑制T2和ILC2的激活	鼠	［97］
志贺氏菌感染	干扰噬菌体感染易感大肠杆菌	大肠杆菌	［98］
空肠弯曲杆菌感染	调节保护性先天免疫相关基因的表达	HT-29细胞	［99］
非传染性葡萄膜炎	调节肠眼轴	鼠	［100］
肠上皮屏障功能障碍	维持与TJ结构相关的亚细胞定位	T-84和Caco-2细胞	［2］
人轮状病毒感染	增强先天和B细胞免疫反应	猪	［77］
结肠癌	通过调节信号通路诱导细胞凋亡	HT-29细胞	［3］
IBD	调节特定mRNA的表达	鼠	［6］
呼吸道合胞病毒	逆转肠道生态失调	鼠	［101］
抗生素不适	阻断病理肠脑回路	鼠	［102］
慢性细菌性前列腺炎	左氧氟沙星联合治疗	人体	［103］
肠道炎症性/感染性腹泻	外膜囊泡调节免疫细胞的功能	RAW 264.7细胞	［104］
苯丙酮尿症	降解丙氨酸	人体	［89］
口腔癌	检测乳酸	人体	［105］

产物 Product	策略 Strategy	产量 Yield	参考文献 Reference
β-丙氨酸	表达天冬氨酸转移酶，增加富马酸的供应，增强前体途径	11.9 g/L	［113］
L-精氨酸	删除负调控基因，插入抗反馈酶基因	-	［123］
3-羟基丁酸	将合酶基因整合基因组	0.6 g/L	［42］
丁酸	异源表达BCD和BUT基因	297 mg/L	［124］
丁酸盐	葡萄糖途径整合丁基辅酶A	20 mmol/L	［125］
5-氨基乙酰丙酸	过表达合酶，抑制降解途径	300 mg/L	［85］
胰高血糖素样肽-1	过表达编码基因	-	［126］
血红素衍生分子	在EcN基因组上整合T7表达系统，表达含血红素蛋白	-	［44］
γ-氨基丁酸	使用无抗质粒作为载体	17.9 g/L	［31］
细菌纤维素	过表达纤维素合酶	1.94 g/L	［117］
Omega-3	过表达合酶基因簇	31.36 mg/L	［119］
肝素前体	强化前体途径，过表达合酶，调节转运蛋白	12.2 g/L	［116］