Acetic acid is a common inhibitor present in lignocellulose hydrolysate,which inhibits the ethanol production by yeast strains.Therefore,the cellulosic ethanol industry requires yeast strains that can tolerate acetic ...Acetic acid is a common inhibitor present in lignocellulose hydrolysate,which inhibits the ethanol production by yeast strains.Therefore,the cellulosic ethanol industry requires yeast strains that can tolerate acetic acid stress.Here we demonstrate that overexpressing a yeast native arginase-encoding gene,CAR1,renders Saccharomyces cerevisiae acetic acid tolerance.Specifically,ethanol yield increased by 27.3%in the CAR1-overexpressing strain compared to the control strain under 5.0 g/L acetic acid stress.The global intracellular amino acid level and compositions were further analyzed,and we found that CAR1 overexpression reduced the total amino acid content in response to acetic acid stress.Moreover,the CAR1 overexpressing strain showed increased ATP level and improved cell membrane integrity.Notably,we demonstrated that the effect of CAR1 overexpression was independent of the spermidine and proline metabolism,which indicates novel mechanisms for enhancing yeast stress tolerance.Our studies also suggest that CAR1 is a novel genetic element to be used in synthetic biology of yeast for efficient production of fuel ethanol.展开更多
目的分离纯化碧根果致敏原Car i 1,并对其结构进行表征鉴定。方法以新鲜碧根果果仁为原料,通过粉碎、脱脂、浸提、粗分级、凝胶过滤层析,对碧根果致敏原蛋白Car i 1进行分离纯化。结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串...目的分离纯化碧根果致敏原Car i 1,并对其结构进行表征鉴定。方法以新鲜碧根果果仁为原料,通过粉碎、脱脂、浸提、粗分级、凝胶过滤层析,对碧根果致敏原蛋白Car i 1进行分离纯化。结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱法和免疫印迹法3种方法对Cari1进行鉴定,并通过圆二色谱仪与紫外分光光度计表征其二、三级结构。结果本方法纯化获得碧根果致敏原Cari1,单轮制备量可达5 mg以上,且纯度大于95%,蛋白质高级结构未被破坏,能够被全部3名碧根果过敏患者的血清准确识别。结论该纯化方法技术路线简单、设备要求低且单次制备量高,总得率可达65%,操作便捷,为碧根果致敏原Car i 1的相关研究奠定了物质基础。展开更多
基金supported financially by National Key Research and Development Program(No.2022YFE0108500)National Natural Science Foundation of China(No.21978168)to X.-Q.ZJFQ appreciates the grant from Sichuan Natural Science Foundation(No.2023NSFSC0132).
文摘Acetic acid is a common inhibitor present in lignocellulose hydrolysate,which inhibits the ethanol production by yeast strains.Therefore,the cellulosic ethanol industry requires yeast strains that can tolerate acetic acid stress.Here we demonstrate that overexpressing a yeast native arginase-encoding gene,CAR1,renders Saccharomyces cerevisiae acetic acid tolerance.Specifically,ethanol yield increased by 27.3%in the CAR1-overexpressing strain compared to the control strain under 5.0 g/L acetic acid stress.The global intracellular amino acid level and compositions were further analyzed,and we found that CAR1 overexpression reduced the total amino acid content in response to acetic acid stress.Moreover,the CAR1 overexpressing strain showed increased ATP level and improved cell membrane integrity.Notably,we demonstrated that the effect of CAR1 overexpression was independent of the spermidine and proline metabolism,which indicates novel mechanisms for enhancing yeast stress tolerance.Our studies also suggest that CAR1 is a novel genetic element to be used in synthetic biology of yeast for efficient production of fuel ethanol.
文摘目的分离纯化碧根果致敏原Car i 1,并对其结构进行表征鉴定。方法以新鲜碧根果果仁为原料,通过粉碎、脱脂、浸提、粗分级、凝胶过滤层析,对碧根果致敏原蛋白Car i 1进行分离纯化。结合十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱法和免疫印迹法3种方法对Cari1进行鉴定,并通过圆二色谱仪与紫外分光光度计表征其二、三级结构。结果本方法纯化获得碧根果致敏原Cari1,单轮制备量可达5 mg以上,且纯度大于95%,蛋白质高级结构未被破坏,能够被全部3名碧根果过敏患者的血清准确识别。结论该纯化方法技术路线简单、设备要求低且单次制备量高,总得率可达65%,操作便捷,为碧根果致敏原Car i 1的相关研究奠定了物质基础。
