Enhancing single-cell hyaluronic acid biosynthesis by microbial morphology engineering

查看全文 作　　者：Yukun [1,2]Zheng;Fangyu [1,2]Cheng;Bo [1,2]Zheng;Huimin [1,2,3]Yu

机构地区：[1]Key Laboratory of Industrial Biocatalysis,Ministry of Education,Beijing,100084,PR China;[2]Department of Chemical Engineering,Tsinghua University,Beijing,100084,PR China;[3]Center for Synthetic and Systems Biology,Tsinghua University,Beijing,100084,PR China

出　　处：《Synthetic and Systems Biotechnology》索引2020年第5卷第4期,共8页

基　　金：This work was supported by National Key R&D Program of China[2018YFA0902200];the National Natural Science Foundation of China[No.21776157]. 摘　　要：Microbial morphology engineering is a novel approach for cell factory to improve the titer of target product in bio-manufacture.Hyaluronic acid(HA),a valuable glycosaminoglycan polymerized by HA synthase(HAS),a membrane protein,is particularly selected as the model product to improve its single-cell HA-producing capacity via morphology engineering.DivIVA and FtsZ,the cell-elongation and cell division related protein,respectively,were both down/up dual regulated in C.glutamicum via weak promoter substitution or plasmid overexpression.Different from the natural short-rod shape,varied morphologies of engineered cells,i.e.small-ellipsoid-like(DivIVA-reduced),bulb-like(DivIVA-enhanced),long-rod(FtsZ-reduced)and dumbbell-like(FtsZ-enhanced),were observed.Applying these morphology-changed cells as hosts for HA production,the reduced expression of both DivIVA and FtsZ seriously inhibited normal cell growth;meanwhile,overexpression of DivIVA didn't show morphology changes,but overexpression of FtsZ surprisingly change the cell-shape into long and thick rod with remarkably enlarged single-cell surface area(more than 5.2-fold-increase).And finally,the single-cell HA-producing capacity of the FtsZ-overexpressed C.glutamicum was immensely improved by 13.5-folds.Flow cytometry analyses verified that the single-cell HAS amount on membrane was enhanced by 2.1 folds.This work is pretty valuable for high titer synthesis of diverse metabolic products with microbial cell factory. 关键词：Morphology engineering Single-cell HA-Producing capacity DivIVA protein FtsZ protein Down/up dual regulation Enlarged cells

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Enhancing single-cell hyaluronic acid biosynthesis by microbial morphology engineering

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