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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2022, Vol. 42 Issue (10): 80-92    DOI: 10.13523/j.cb.2204049
综述     
合成芳香族化合物的酵母底盘改造策略*
李慧敏,贾斌,李霞,刘夺**()
天津大学化工学院 教育部合成生物学前沿科学中心 系统生物工程教育部重点实验室 天津 300072
Advances in Engineering Yeast Chassis for Producing Aromatic Compounds
Hui-min LI,Bin JIA,Xia LI,Duo LIU**()
Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering of Ministry of Education,School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
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摘要：

芳香族化合物种类丰富,在多个行业具有广泛的用途,需求量大。通过构建微生物细胞工厂合成芳香族化合物具有独特的优势和工业化应用前景,其中酵母底盘因其清晰的遗传背景、完善的基因操作工具以及成熟的工业发酵体系等优势,常被用于构建细胞工厂。目前改造酵母底盘生产芳香族化合物的研究取得了一系列进展,并针对关键问题提出了一些可行的解决策略。针对酵母合成芳香族化合物的策略与挑战,从芳香族化合物合成路径改造、多样化碳源利用及转运系统改造、基因组多靶点改造、特殊酵母底盘及混菌系统构建、合成生物学高通量技术的应用这五个方面进行系统地梳理和阐述,为生产芳香族化合物的酵母底盘构建与改造提供思路。
关键词： 芳香族化合物;  酵母;  底盘改造;  基因组重排;  合成生物学    
Abstract:

Aromatic compounds including lots of species are in high demand and widely used in many areas. The synthesis of aromatic compounds by constructing microbial cell factories has unique advantages and industrial applications. Yeast chassis is often used to build cell factories due to its clear genetic background, well-performed gene manipulation tools, and mature industrial system. A series of advances has been made in the production of aromatic compounds by engineering yeast chassis, and several feasible solutions have been proposed to solve key problems. In view of the strategies and challenges of yeast synthesis of aromatic compounds, this paper systematically reviews and elaborates on the five aspects, namely, the aromatic compound synthetic pathway modification, utilization of diverse carbon sources and modification of transport systems, genome multi-target modification, special yeast chassis and microbial consortium construction, and applications of synthetic biology high-throughput technologies, and provides ideas for the construction and engineering of yeast chassis for producing aromatic compounds.
Key words: Aromatic compounds    Yeast    Chassis engineering    Genome rearrangement    Synthetic biology
收稿日期: 2022-04-21 出版日期: 2022-11-04
ZTFLH:  Q819  
基金资助: * 国家自然科学基金青年项目(21706186)
通讯作者: 刘夺     E-mail: duo_liu0@tju.edu.cn
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引用本文:

李慧敏,贾斌,李霞,刘夺. 合成芳香族化合物的酵母底盘改造策略*[J]. 中国生物工程杂志, 2022, 42(10): 80-92.

Hui-min LI,Bin JIA,Xia LI,Duo LIU. Advances in Engineering Yeast Chassis for Producing Aromatic Compounds. China Biotechnology, 2022, 42(10): 80-92.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2204049    或    https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I10/80

图1  产芳香族化合物的酵母底盘改造策略进展
产品 代谢工程策略 发酵方式 产量 文献
酪醇 敲除PDC1、PHA2;表达ARO4K229L、ARO7G141S、ARO3K222L;过表达RKI1和TKL1 摇瓶 702.30 mg/L [7]
5 L发酵罐 9.90 g/L
酪醇 表达EcAROGD146N、EcpheAfbr、EcydiB、EcAROL;过表达TKL1、ARO10;敲除ARO9 摇瓶 541 mg/L [11]
红景天苷 敲除PDC1、PHA2;表达ARO4K229L、ARO7G141S、RrU8GT33opt、ARO3K222L;过表达RKI1和TKL1 摇瓶 1 575.45 mg/L [7]
5 L发酵罐 26.55 g/L
莽草酸 过表达ARO1D920A、ARO4K229L、TKL1 摇瓶 358 mg/L [8]
对香豆酸 过表达ARO4K229L、ARO7G141S、FjTAL、EcAROL;敲除ARO10、PDC5 发酵罐 1.93 g/L [10]
对香豆酸 表达AtPAL2、AtC4H、AtATR2、CYB5、FjTAL、EcAROL、MtPDH1;过表达ARO4K229L、ARO7G141S;敲除PDC5、ARO10 发酵罐 12.5 g/L [5]
对香豆酸 解脂耶氏酵母:敲除YlPYK、YlTYR1、YlTRP2、YlTRP3、YlARO8、YlARO9;过表达ScARO4K229L、AROGS180F、BbXFPK、AcXPK等 摇瓶 593 mg/L [52]
2-苯乙醇 表达EcAROGD146N、EcpheAfbr、EcydiB、EcAROL;过表达TKL1、ARO10;敲除ARO9 摇瓶 643 mg/L [11]
2-苯乙醇 过表达GLN3改造转运系统 摇瓶 3.59 g/L [31]
产品 代谢工程策略 发酵方式 产量 文献
2-苯乙醇 过表达GAP1、ARO8、ARO10、ADH2、GDH2 5 L发酵罐 6.3 g/L [32]
2-苯乙醇 解脂耶氏酵母:过表达YlPAR4、YlARO10、YlARO7、YlPHA2、ScARO7G141S、ScARO4K229L、AROGS180F、BbXFPK、AcXPK等;敲除YlTYR1、YlTRP2、YlTRP3、YlARO8、YlARO9、YlPYK 摇瓶 2 426 mg/L [52]
黏糠酸 敲除ARO3、ARO4、ZWF1;过表达ARO4K229L、EcL_01944opt、Pa5_5120opt、CaHQD2opt、TKL1;生物传感器、定向进化、ARO1截短表达 摇瓶 500 mg/L [14]
发酵罐 2.1 g/L
柚皮素 敲除ARO3、ARO10、PDC5、PDC6;表达ARO4G226S;过表达AtPAL1、CoC4H、CoCPR1、AtCHI1、AtCHS3、CoCHS3、At4CL3、CoTAL1 2 L发酵罐 112 mg/L [16]
柚皮素 解脂耶氏酵母:过表达RtTAL、Pc4CL、PhCHS、MsCHI、GhF3'H、CrCPR、SlF3H、YlARO1、YlACC1、YlACS2;培养条件优化 摇瓶 252 mg/L [51]
圣草酚 表达Pc4CL、PhCHS、MsCHI、AmF3H、AtFLS 摇瓶 134.2 mg/L
花旗松素 表达Pc4CL、PhCHS、MsCHI、AmF3H、AtFLS、PhFMO、CrCPR 摇瓶 110.5 mg/L
天竺葵素-3-
O-葡萄糖苷		 表达HaCHS、MsCHI、At4CL2、AtPAL2、AmC4H、ScCPR1、PhANS、MdF3H、AaDFR、DcA3GT 深孔板 0.85 mg/L [18]
花青素-3-
O-葡萄糖苷		 表达HaCHS、MsCHI、At4CL2、AtPAL2、AmC4H、ScCPR1、PhANS、MdF3H、PhF3'H、PtDFR、AtCPR1、FaA3GT2 深孔板 1.55 mg/L
飞燕草素-3-
O-葡萄糖苷		 表达HaCHS、MsCHI、At4CL2、AtPAL2、AmC4H、ScCPR1、PhANS、SlF3'5'H、MdF3H、IhDFR、AtCPR1、FaA3GT2 深孔板 1.86 mg/L
咖啡酸 表达PaHpaB、SeHpaC、RtTAL 摇瓶 289 mg/L [19]
咖啡酸 敲除ARO10、PDC5;表达ARO7fbr、ARO4fbr、CoTAL、CoC3H、CoCPR1 摇瓶 11.432 mg/L [20]
咖啡酸 敲除ARO10、PDC5、gpp1;过表达ScARO7G141S、ARO4K229L、EcAROL、FjTAL、SbPAL1、AtCPR1、PtrC4H2、PtrC4H1、PtrC3H3、PaHpaB、SeHpaC、ScARO3、ScTAL1、ScTKL1、CkPTA、LmXFPK、ScPHA2、MtPDH1、BiFADS、BsRIBBA等 摇瓶 1.6 g/L [21]
发酵罐 5.5 g/L
阿魏酸 敲除ARO10、PDC5、gpp1;过表达ScARO7G141S、ARO4K229L、EcAROL、FjTAL、SbPAL1、AtCPR1、PtrC4H2、PtrC4H1、PtrC3H3、PaHpaB、SeHpaC、ScARO3、ScTAL1、ScTKL1、CkPTA、LmXFPK、NtCOMT1、ScMET6、LiMETK1、ScADO1、ScSAH1等 摇瓶 0.42 g/L
发酵罐 3.8 g/L
番茄枝碱 表达BvCYP76AD1、PsNCS、6OMT、CNMT、4'OMT、NMCH 摇瓶 80.6 μg/L [22]
对氨基苯甲酸 表达ABZ1、 ABZ2、ARO4K229L;敲除ARO7、TRP3 发酵罐 215 mg/L [27]
原儿茶酸 敲除ADH6、ADH7、BDH2、FDC、MHT1、SAM4;表达4CL、Ppech、vdh、pobA、PdvanA、PdvanB、ligM;过表达MET6、Rdmetf1 摇瓶 810 mg/L [28]
姜黄素 敲除fdc1;过表达PpFerA、ClDCS、ClCURS 摇瓶 2.7 mg/L [30]
多巴胺 CYP76AD1W13L, F309L 摇瓶 10.8 mg/L [67]
紫色杆菌素 SCRaMbLE-in 摇瓶 17.5 mg/L [48]
紫色杆菌素 解脂耶氏酵母:过表达ScARO4K229L、AROGS180F、BbXFPK、AcXPK等;敲除YlTYR1、YlTRP2、YlTRP3、YlARO8、YlARO9、YlPYK 摇瓶 366 mg/L [52]
脱氧紫色杆菌素 摇瓶 55 mg/L
白藜芦醇 过表达ARO4K229L、ScARO7G141S、ScAcc1S659A、 S1157A、HaTAL、At4CL1、VvVST1 摇瓶 415.65 mg/L
或531.41 mg/L		 [9]
白藜芦醇 表达AtPAL2、AtC4H、At4CL2、VvVST1 摇瓶 151.65 mg/L [29]
白藜芦醇 解脂耶氏酵母;过表达FjTAL、At4CL1、VvVST1、YlARO4K221L、YlARO7G139S;敲除PDC5 发酵罐 12.4 g/L [54]
表1  酵母合成芳香族化合物所采用的底盘改造策略
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