Engineering Nannochloropsis oceanica for the production of diterpenoid compounds  

作　　者：Zhi-Yan Du Wajid W.Bhat Eric Poliner Sean Johnson Conor Bertucci Eva Farre Bjoern Hamberger 

机构地区：[1]Department of Molecular Biosciences and Bioengineering,University of Hawaii at Manoa,Honolulu,Hawaii,USA [2]Department of Biochemistry and Molecular Biology,Michigan State University,East Lansing,Michigan,USA [3]Department of Plant Biology,Michigan State University,East Lansing,Michigan,USA [4]New England Biolabs Inc.,240 County Roadlpswich,01938 MA,USA

出　　处：《mLife》2023年第4期428-437,共10页微生物（英文）

基　　金：U.S.Department of Agriculture National Institute of Food and Agriculture (HATCH project HAW05047-H and 2023-70003-38783);Center for Tropical and Subtropical Aquaculture (Grant No.2020-38500-32559);the National Science Foundation (Grant Number 2121410);the US Department of Energy-Great Lakes Bioenergy Research Center Cooperative Agreement DE-SC0018409,start-up funding from the Department of Molecular Biology and Biochemistry;support by the USDA National Institute of Food and Agriculture,HATCH project MICL02454.B.H;supported by the National Science Foundation (Grant Number 1737898);E.M.F acknowledges funding from the National Science Foundation (IOS-134721).

摘　　要：Photosynthetic microalgae like Nannochloropsis hold enormous potential as sustainable,light-driven biofactories for the production of high-value natural products such as terpenoids.Nannochloropsis oceanica is distinguished as a particularly robust host with extensive genomic and transgenic resources available.Its capacity to grow in wastewater,brackish,and sea waters,coupled with advances in microalgal metabolic engineering,genome editing,and synthetic biology,provides an excellent opportunity.In the present work,we demonstrate how N.oceanica can be engineered to produce the diterpene casbene—an important intermediate in the biosynthesis of pharmacologically relevant macrocyclic diterpenoids.Casbene accumulated after stably expressing and targeting the casbene synthase from Daphne genkwa(DgTPS1)to the algal chloroplast.The engineered strains yielded production titers of up to 0.12 mg g^(−1) total dry cell weight(DCW)casbene.Heterologous overexpression and chloroplast targeting of two upstream rate-limiting enzymes in the 2-C-methyl-d-erythritol 4-phosphate pathway,Coleus forskohlii 1-deoxy-d-xylulose-5-phosphate synthase and geranylgeranyl diphosphate synthase genes,further enhanced the yield of casbene to a titer up to 1.80 mg g^(−1) DCW.The results presented here form a basis for further development and production of complex plant diterpenoids in microalgae.

关 键 词：casbene MICROALGAE plastid targeting synthetic biology terpene synthase 

分 类 号：Q938.8[生物学—微生物学]