Patatin,a prominent food protein derived from potatoes,is renowned for its exceptional nutritional value.Patatin has been characterized for its diverse physiological attributes,including esterase activity,antioxidativ...Patatin,a prominent food protein derived from potatoes,is renowned for its exceptional nutritional value.Patatin has been characterized for its diverse physiological attributes,including esterase activity,antioxidative properties,cholesterol-lowering effects,and high lysine content,alongside notable physicochemical traits such as foaming,emulsification,and gelation capabilities.Conventional methods for patatin extraction are fraught with inefficiencies,elevated costs,and detrimental impacts on protein structural and functional integrity.Herein,we leveraged an optimized strategy integrating an expression cassette toolbox and regulation of protein secretion to harness Komagataella phaffii as the expression host and achieved an expression level of 3.2 g per litre(g/L)in a 5-Litre bioreactor,which is the highest yield of patatin production using engineered bacteria and funguses that has been reported thus far.In this study,we innovatively refined the endogenous promoter PCAT1,and its efficacy in driving heterologous protein expression under methanol induction surpassed that of the conventional AOX1 promoter.Furthermore,crucial nodes for patatin heterologous expression in yeast were identified,substantially curtailing the production costs associated with patatin synthesis.展开更多
The transcription factor Ets1 is essential for the development of invariant natural killer T(iNKT)cells;however,its detailed role and mechanism of action are unknown.Here,we show that Ets1 is dispensable for CD1d-medi...The transcription factor Ets1 is essential for the development of invariant natural killer T(iNKT)cells;however,its detailed role and mechanism of action are unknown.Here,we show that Ets1 is dispensable for CD1d-mediated selection,but is essential for subsequent differentiation of post-selected iNKT cells.This function is partly compensated by forced expression of a transgenic Vα14 Jα18 TCR and is independent of its N-terminal Pointed(PNT)domain.Ets1 is also required for optimal expression of cytokines by differentiated iNKT1 and iNKT2 cells,but inhibits the differentiation/function of iNKT17 in vitro and in vivo.These functions,however,depend on the PNT domain.Taken together,our results indicate that Ets1 regulates the differentiation/function of iNKT cells at multiple steps through both PNT domaindependent and PNT domain-independent mechanisms.展开更多
基金supported by the National Key Research and Devel-opment Program(2021YFC2104000)the National Natural Science Foundation of China(32272276)the Fundamental Research Funds for the Central Universities.
文摘Patatin,a prominent food protein derived from potatoes,is renowned for its exceptional nutritional value.Patatin has been characterized for its diverse physiological attributes,including esterase activity,antioxidative properties,cholesterol-lowering effects,and high lysine content,alongside notable physicochemical traits such as foaming,emulsification,and gelation capabilities.Conventional methods for patatin extraction are fraught with inefficiencies,elevated costs,and detrimental impacts on protein structural and functional integrity.Herein,we leveraged an optimized strategy integrating an expression cassette toolbox and regulation of protein secretion to harness Komagataella phaffii as the expression host and achieved an expression level of 3.2 g per litre(g/L)in a 5-Litre bioreactor,which is the highest yield of patatin production using engineered bacteria and funguses that has been reported thus far.In this study,we innovatively refined the endogenous promoter PCAT1,and its efficacy in driving heterologous protein expression under methanol induction surpassed that of the conventional AOX1 promoter.Furthermore,crucial nodes for patatin heterologous expression in yeast were identified,substantially curtailing the production costs associated with patatin synthesis.
基金supported by grants from the United States Department of Defense(W81XWH-11-1-0492 to I.-C.H.)the National Institutes of Health(AR070171 to I.-C.H.and HL082717 to P.O.)+1 种基金the Ministry of Science and Technology(103-2311-B-650-001 and 105-2311-B-214-002 to T.-S.T.),TaiwanEDa Hospital(103-EDN16,104-EDN12,105-EDN07,and EDAHI105002 to T.-S.T.),Taiwan.
文摘The transcription factor Ets1 is essential for the development of invariant natural killer T(iNKT)cells;however,its detailed role and mechanism of action are unknown.Here,we show that Ets1 is dispensable for CD1d-mediated selection,but is essential for subsequent differentiation of post-selected iNKT cells.This function is partly compensated by forced expression of a transgenic Vα14 Jα18 TCR and is independent of its N-terminal Pointed(PNT)domain.Ets1 is also required for optimal expression of cytokines by differentiated iNKT1 and iNKT2 cells,but inhibits the differentiation/function of iNKT17 in vitro and in vivo.These functions,however,depend on the PNT domain.Taken together,our results indicate that Ets1 regulates the differentiation/function of iNKT cells at multiple steps through both PNT domaindependent and PNT domain-independent mechanisms.
