文献类型：期刊文献

英文题名：Enhancing Trans-Nerolidol Productivity in Yarrowia lipolytica by Improving Precursor Supply and Optimizing Nerolidol Synthase Activity

作者：Liu, Feng[1];Liu, Shun-Cheng[1,2];Qi, Yi-Ke[1];Liu, Zhijie[3];Chen, Jun[1];Wei, Liu-Jing[1];Hua, Qiang[1,4]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]North China Univ Sci & Technol, Sch Basic Med Sci, Hebei Key Lab Chron Dis, Tangshan Key Lab Preclin & Basic Res Chron Dis, Tangshan 063210, Peoples R China;[3]Hubei Univ Technol, Minist Educ & Hubei Prov, Cooperat Innovat Ctr Ind Fermentat, Key Lab Fermentat Engn,Minist Educ, Wuhan 430068, Peoples R China;[4]Shanghai Collaborat Innovat Ctr Biomfg Technol, Shanghai 200237, Peoples R China

年份：2022

卷号：70

期号：48

起止页码：15157

外文期刊名：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY

收录：;EI(收录号：20225013226701);WOS:【SCI-EXPANDED(收录号:WOS:000891457800001)】；

基金：? ACKNOWLEDGMENTS This study was supported by the National Key R&D Program of China (2017YFE0115600, 2019YFA0904302) , the National Natural Science Fund for Young Scholars (32001035) , and the Hebei Province National Natural Science Fund for Young Scholars (B2022209030) .

语种：英文

外文关键词：trans-nerolidol; protein engineering; peroxisomal acetyl-CoA pool; pathway compartmentation; Yarrowia lipolytica

摘要：The low enzymatic capability of terpene synthases and the limited availability of precursors often hinder the productivity of terpenes in microbial hosts. Herein, a systematic approach combining protein engineering and pathway compartmentation was exploited in Yarrowia lipolytica for the high-efficient production of trans-nerolidol, a sesquiterpene with various commercial applications. Through the single-gene overexpression, the reaction catalyzed by nerolidol synthase (FaNES1) was identified as another rate-limiting step. An optimized FaNES1G498Qwas then designed by rational protein engineering using homology modeling and docking studies. Additionally, further improvement of trans-nerolidol production was observed as enhancing the expression of an endogenous carnitine acetyltransferase (CAT2) putatively responsible for acetyl-CoA shuttling between peroxisome and cytosol. To harness the peroxisomal acetyl-CoA pool, a parallel peroxisomal pathway starting with acetylCoA to trans-nerolidol was engineered. Finally, the highest reported titer of 11.1 g/L trans-nerolidol in the Y. lipolytica platform was achieved in 5 L fed-batch fermentation with the carbon restriction approach.