Overexpression of the Key Enzymes in the Methylerythritol 4-phosphate Pathway in Corynebacterium glutamicum for Improving Farnesyl Diphosphate-Derived Terpene Production
Overexpression of the Key Enzymes in the Methylerythritol 4-phosphate Pathway in Corynebacterium glutamicum for Improving Farnesyl Diphosphate-Derived Terpene Production
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DOI：
10.1021/acs.jafc.0c04307
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发表时间：
2020-09-30
影响因子：
6.1
通讯作者：
Woo, Han Min
中科院分区：
文献类型：
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作者：
Lim, Hyeonbae;Park, Jaehyun;Woo, Han Min
采用系统和组合优化的方法对微生物进行代谢工程，以确定过表达的关键基因靶点，从而增加萜类化合物生产的中间库。本研究以工业宿主谷氨酸棒杆菌中的甲基赤藓糖醇4-磷酸(MEP)途径为研究对象，以确定其过度表达将促进法尼基二磷酸(FPP)衍生的萜类化合物(角鲨烯和α-法尼烯)产生的关键基因。在高通量发酵中，使用MEP途径中基因的单一、双重和三重表达的组合方法，过表达ispDF基因以及已知的DXS和IDI基因，在增加角鲨烯含量方面最有效，即增加14倍。Dxr基因被确定为生产α-法尼烯的关键靶标酶。这一结果可以为通过优化的MEP途径改进谷氨酸菌代谢工程以生产萜类提供基础信息。
A systematic and combinatorial optimization has been employed to metabolically engineer microbes for identifying key gene targets for overexpression to increase the intermediate pools for terpenoid production. Herein, the methylerythritol 4-phosphate (MEP) pathway in Corynebacterium glutamicum, an industrial host, was investigated to identify the key genes whose overexpression would improve the production of farnesyl diphosphate (FPP)-derived terpenoids (squalene and alpha-farnesene). Using a combinatorial approach with the single, double, and triple expression of genes in the MEP pathway in a high-throughput fermentation, overexpression of the ispDF genes, along with the known dxs and idi genes, was most effective at increasing the squalene contents, i.e., by 14-fold. The dxr gene was identified as the key target enzyme for alpha-farnesene production. This result could provide fundamental information for improving the metabolic engineering of C. glutamicum for terpene production via an optimized MEP pathway.