微生物合成2-苯乙醇研究进展<sup>*</sup>

doi:10.13523/j.cb.2203073

中国生物工程杂志

2022, Vol. 42

Issue (8): 128-136 DOI: 10.13523/j.cb.2203073

综述

微生物合成2-苯乙醇研究进展^*

卞一凡,刘姝晗,张贝萌,张玉龙,李辛桐,王鹏超^**(

)

东北林业大学生命科学学院哈尔滨 150000

Advances in Microbial Synthesis of 2-Phenylethanol

BIAN Yi-fan,LIU Shu-han,ZHANG Bei-meng,ZHANG Yu-long,LI Xin-tong,WANG Peng-chao^**(

)

School of Life Science, Northeast Forestry University, Harbin 150000, China

全文: PDF(1688 KB) HTML

摘要：

2-苯乙醇(2-PE)是一种具有广阔应用前景的高级芳香醇。由于化学合成的复杂性和天然提取的高昂成本,近年来,利用微生物发酵合成2-PE受到广泛关注。许多微生物有天然合成2-PE的能力,但产量相对较低,并不适合大规模生产。在最近几年的研究中,利用代谢工程和合成生物学技术,通过上调限速酶基因表达水平,改善前体转运,提高 2-PE耐受性等多方面优化,2-PE的微生物产量有了大幅度的提高。综述微生物合成2-PE的相关研究进展,分析关键代谢调控的机制,并就目前存在的问题提出了改进建议。

关键词： 2-苯乙醇; 生物合成; 代谢工程; 酵母; 大肠杆菌

Abstract:

2-Phenylethanol (2-PE) is an important aromatic with broad application prospects. Due to the complexity of chemical synthesis and the high cost of natural extraction, the synthesis of 2-PE by microbial fermentation has gained extensive attention recently. Various microorganisms have the ability to synthesize 2-PE naturally with low yield. In recent years, using genetic engineering and synthetic biology technology, the microbial yield of 2-PE has been greatly improved by up-regulating the expression level of rate-limiting enzyme gene, improving precursors transport and enhancing the tolerance of 2-PE. The research progress of microbial synthesis of 2-PE is reviewed, the mechanism of key metabolic regulation is analyzed, current problems are addressed, and suggestions for improvement are proposed.

Key words: 2-Phenylethanol Biosynthesis Metabolic engineering Yeast Escherichia coli

收稿日期: 2022-03-31 出版日期: 2022-09-07

ZTFLH:

Q36

基金资助: * 国家级大学生创新创业项目(202110225006)

通讯作者: 王鹏超 E-mail: pengchaowang1990@nefu.edu.cn

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引用本文:

卞一凡,刘姝晗,张贝萌,张玉龙,李辛桐,王鹏超. 微生物合成2-苯乙醇研究进展^*[J]. 中国生物工程杂志, 2022, 42(8): 128-136.

BIAN Yi-fan,LIU Shu-han,ZHANG Bei-meng,ZHANG Yu-long,LI Xin-tong,WANG Peng-chao. Advances in Microbial Synthesis of 2-Phenylethanol. China Biotechnology, 2022, 42(8): 128-136.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2203073 或 https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I8/128

图1 微生物 2-PE 合成途径及优化靶点

表1 苯丙酮酸途径中部分酶活汇总

优化方向	菌株	合成途径	策略	底物	产量	得率 /(g/g)	参考文献
提高L-Phe 的利用率	耶罗维解脂酵母 Yarrowia lipolytica	艾氏途径	引入L-Phe通透酶Gap1的同源物GapY3,并过表达合成L-Phe转氨基受体α-酮戊二酸的乌头酸水合酶及线粒体2-氧代二羧酸载体odc1	L-Phe、葡萄糖	2.67 g/L	0.702	[10]
	酿酒酵母S. cerevisiae	艾氏途径	过表达链霉菌的L-谷氨酸氧化酶strlgox	L-Phe、葡萄糖	4.02 g/L	0.6	[11]
抑制竞争途径	耶罗维解脂酵母 Yarrowia lipolytica	艾氏途径	敲除乙醛脱氢酶ald2和ald3,并敲除二酰甘油酰基转移酶抑制脂肪酸合成	L-Phe、葡萄糖	2.67 g/L	0.702	[10]
	酿酒酵母S. cerevisiae	艾氏途径	利用CRISPR/Cas9敲除乙酰基转移酶	L-Phe、葡萄糖	1.2 g/L	—	[13]
	巴斯德毕赤酵母 Pichia pastoris	苯丙酮酸途径	更换弱的tyr1启动子,下调其表达水平	葡萄糖	1.17 g/L	0.059	[15]
	大肠杆菌E.coli	苯乙烯衍生途径	敲除PykF、PykA和Ccr,并敲除feaB,减少副产物苯乙酸	L-Phe	1.16 g/L	0.039	[22]
上调关键酶的表达	耶罗维解脂酵母 Yarrowia lipolytica	艾氏途径	上调苯丙酮酸脱羧酶aro10和苯乙醛还原酶PAR4	L-Phe、葡萄糖	2.67 g/L	0.702	[10]
	酿酒酵母 S. cerevisiae	艾氏途径	引入乳酸乳杆菌的苯丙酮酸脱羧酶kdcA	L-Phe、葡萄糖	4.02 g/L	0.6	[11]
	大肠杆菌E.coli	艾氏途径	引入玫瑰中的苯乙醛合成酶PAAS	L-Phe	0.39 g/L	0.49	[21]
	酿酒酵母S. cerevisiae	苯丙酮酸途径	过表达转酮醇酶基因TKL1,并在此基础上上调突变的磷酸烯醇式丙酮酸羧激酶PYK1	葡萄糖	1.53 g/L	0.077	[14]
	巴斯德毕赤酵母 Pichia pastoris	苯丙酮酸途径	3-脱氧基-7-脱氧庚磺酸-7-合酶合酶AroG和分支酸变位酶/预苯酸脱水酶PheA的过表达	葡萄糖	1.17 g/L	0.059	[15]
	大肠杆菌E.coli	苯丙酮酸途径	过表达预苯酸脱水酶(PheA)和DAHP合成酶(AroF)	葡萄糖	0.33 g/L	—	[24]
	大肠杆菌E.coli	苯丙酮酸途径	过表达PEP合酶ppsA和转酮酶tktA	葡萄糖	0.32 g/L	0.053	[25]
	大肠杆菌E.coli	苯乙烯衍生途径	引入外源基因pal2、fdc1和styAB,并过表达氧化苯乙烯异构酶	L-Phe	1.16 g/L	0.039	[22]
	无细胞系统	苯乙烯衍生途径	共表达苯丙氨酸解氨酶pal2和苯丙酸脱羧酶pad	L-Phe	0.20 g/L	—	[23]
降低2-PE 的细胞毒性	季也蒙酵母 Meyerozyma guilliermondii	艾氏途径	自身具有高耐受性;膜蛋白中插入酶、热休克蛋白Hsp90和伴侣蛋白SGT1	L-Phe	3.1 g/L	0.39	[32]
	鲁氏酵母Zygosaccha- romyces rouxii	艾氏途径/ 苯丙酮酸途径	自身具有高耐受性,过表达aro8和aro10	L-Phe/ 葡萄糖	3.58 g/L 3.8 mg/L	0.86	[7]
	甘油假丝酵母 Candida glycerinogenes	艾氏途径	自身具有高耐受性	L-Phe	5.0 g/L	0.71	[33]
	地衣芽孢杆菌 Bacillus licheniformis	艾氏途径	自身具有高耐受性;共表达乳酸乳杆菌的kivD和大肠杆菌的yghD	L-Phe	3.04 g/L	0.61	[29]
	酿酒酵母S. cerevisiae	艾氏途径	使用油酸进行萃取	L-Phe	12.6 g/L	0.32	[6,36]

表2 微生物合成2-PE优化途径及策略汇总

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