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 membra...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.展开更多
基金This work was supported by National Key R&D Program of China[2018YFA0902200]the National Natural Science Foundation of China[No.21776157].
文摘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.
