Biosynthesis of a-bisabolene from low-cost renewable feedstocks by peroxisome engineering and systems metabolic engineering of the yeast Yarrowia lipolytica

被引：16

作者：

Zhao, Baixiang ^{[1
]}

Zhang, Yahui ^{[1
]}

Wang, Yaping ^{[1
]}

Lu, Zhihui ^{[1
]}

Miao, Lin ^{[1
]}

Wang, Shuhui ^{[1
]}

Li, Zhuo ^{[1
]}

Sun, Xu ^{[1
]}

Han, Yuqing ^{[1
]}

He, Sicheng ^{[1
]}

Zhang, Ziyuan ^{[1
]}

Xiao, Dongguang ^{[1
]}

Zhang, Cuiying ^{[1
]}

Foo, Jee Loon ^{[2
,3
,4
]}

Wong, Adison ^{[5
]}

Yu, Aiqun ^{[1
]}

机构：

[1] Tianjin Univ Sci & Technol, State Key Lab Food Nutr & Safety, Key Lab Ind Fermentat Microbiol, Tianjin Key Lab Ind Microbiol,Coll Biotechnol,Mini, 29 13th St TEDA, Tianjin 300457, Peoples R China

[2] Natl Univ Singapore, Yong Loo Lin Sch Med, Synthet Biol Translat Res Programme, Singapore 119228, Singapore

[3] Natl Univ Singapore, NUS Synthet Biol Clin & Technol Innovat SynCTI, Singapore 117456, Singapore

[4] Natl Univ Singapore, Yong Loo Lin Sch Med, Dept Biochem, S-117597 Singapore, Singapore

[5] Singapore Inst Technol, Food Chem & Biotechnol Cluster, Singapore 138683, Singapore

来源：

GREEN CHEMISTRY | 2023年 / 25卷 / 20期

关键词：

MULTIDRUG EFFLUX PUMPS; BETA-OXIDATION; ACETYL-COA; PATHWAY;

D O I：

10.1039/d3gc01936e

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Establishing efficient synthetic pathways for microbial production of biochemicals is often hampered by competing pathways and insufficient precursor supply. Compartmentalization in cellular organelles can isolate biosynthetic pathways from competing pathways, and thus provide a more compact and conducive environment for biosynthesis. Herein, the farnesyl diphosphate biosynthetic pathway and a-bisabolene synthase were compartmentalized in the yeast Yarrowia lipolytica peroxisome to enable high-level a-bisabolene production. Along with compartmentalization of the a-bisabolene biosynthesis pathway, a systems metabolic engineering approach that comprises mediating product export with an efflux pump, optimizing the gene copy numbers of rate-limiting enzymes, balancing the distribution of the common precursor acetyl-CoA between native lipid biosynthesis and heterologous a-bisabolene production, improving ATP and acetyl-CoA supply, and dynamic regulation of peroxisomes was then employed to further enhance the bisabolene production. Consequently, the optimized engineered strain produced 3028.9 mg L-1 a-bisabolene from waste cooking oil as the sole carbon source through shake flask cultivation. Finally, fed-batch fermentation was performed to accomplish a-bisabolene production at 15.5 g L-1, the highest ever reported for a-bisabolene from an engineered microbe. Taken together, this work lays the foundation for peroxisome engineering of Y. lipolytica, towards the production of a-bisabolene in a highly sustainable manner.

引用

页码：8145 / 8159

页数：15