Efficient conversion of corn stover to bioethanol via simultaneous saccharification and fermentation(SSF)is a promising strategy for sustainable biofuel production.A major current barrier to this process is the limite...Efficient conversion of corn stover to bioethanol via simultaneous saccharification and fermentation(SSF)is a promising strategy for sustainable biofuel production.A major current barrier to this process is the limited ther-motolerance of Saccharomyces cerevisiae,which hampers its performance under the high-temperature conditions required for efficient SSF.In this study,we identified TrRCC1,a gene from Trichoderma reesei,as a candidate for improving microbial stress resistance.Overexpression of TrRCC1 in both T.reesei Rut C30 and S.cerevisiae BY4741 significantly enhanced thermotolerance.In T.reesei Rut C30,TrRCC1 overexpression improved heat resistance and increased cellulase production by 2.5-fold compared to the wild-type strain.In S.cerevisiae BY4741,TrRCC1 overexpression resulted in enhanced thermotolerance and a 21.8%increase in ethanol production during SSF of corn stover.The ethanol concentration achieved in the SSF process with TrRCC1-overexpressing S.cerevisiae was 44.1 g/L,which was a notable improvement over control strain production.These findings highlight the potential of TrRCC1 as a key gene for engineering microbial strains with improved stress resistance to enhance the efficiency of bioethanol production from lignocellulosic biomass.展开更多
通过对香菇C91-3转录本Unigene 24277基因的生物信息学分析,克隆表达含RCC1结构域的Unigene24277基因,并研究其抗肿瘤活性。从香菇C91-3菌丝体中提取总RNA,反转录合成c DNA,并利用Rapid Amplification of c DNA Ends(RACE)技术获得基因...通过对香菇C91-3转录本Unigene 24277基因的生物信息学分析,克隆表达含RCC1结构域的Unigene24277基因,并研究其抗肿瘤活性。从香菇C91-3菌丝体中提取总RNA,反转录合成c DNA,并利用Rapid Amplification of c DNA Ends(RACE)技术获得基因全长。NCBI数据库分析提示其含有RCC1结构域。PCR扩增RCC1结构域,将其克隆产物与p ET-32a(+)载体连接,热转化至E.coil Rosetta-gami(DE3)中诱导表达,纯化、复性后,通过MTT法研究其抗肿瘤活性。结果显示原核表达载体构建成功,重组蛋白成功诱导表达,并初步证明了重组蛋白具有抑制肿瘤细胞增殖活性的功能,为后续抗肿瘤机制的探究奠定了基础。展开更多
基金supported by the National Natural Science Foun-dation of China(22378033)Natural Science Foundation Project of Chongqing,the Chongqing Science and Technology Commis-sion(CN)(CSTB2022NSCQ-MSX0544)+1 种基金Science and Technology Re-search Program of Chongqing Municipal Education Commission(KJZD-M202401502 and KJQN202301546)Postgraduate Research and Innovation Project of Chongqing University of Science and Technology(YKJCX2420531).
文摘Efficient conversion of corn stover to bioethanol via simultaneous saccharification and fermentation(SSF)is a promising strategy for sustainable biofuel production.A major current barrier to this process is the limited ther-motolerance of Saccharomyces cerevisiae,which hampers its performance under the high-temperature conditions required for efficient SSF.In this study,we identified TrRCC1,a gene from Trichoderma reesei,as a candidate for improving microbial stress resistance.Overexpression of TrRCC1 in both T.reesei Rut C30 and S.cerevisiae BY4741 significantly enhanced thermotolerance.In T.reesei Rut C30,TrRCC1 overexpression improved heat resistance and increased cellulase production by 2.5-fold compared to the wild-type strain.In S.cerevisiae BY4741,TrRCC1 overexpression resulted in enhanced thermotolerance and a 21.8%increase in ethanol production during SSF of corn stover.The ethanol concentration achieved in the SSF process with TrRCC1-overexpressing S.cerevisiae was 44.1 g/L,which was a notable improvement over control strain production.These findings highlight the potential of TrRCC1 as a key gene for engineering microbial strains with improved stress resistance to enhance the efficiency of bioethanol production from lignocellulosic biomass.
文摘通过对香菇C91-3转录本Unigene 24277基因的生物信息学分析,克隆表达含RCC1结构域的Unigene24277基因,并研究其抗肿瘤活性。从香菇C91-3菌丝体中提取总RNA,反转录合成c DNA,并利用Rapid Amplification of c DNA Ends(RACE)技术获得基因全长。NCBI数据库分析提示其含有RCC1结构域。PCR扩增RCC1结构域,将其克隆产物与p ET-32a(+)载体连接,热转化至E.coil Rosetta-gami(DE3)中诱导表达,纯化、复性后,通过MTT法研究其抗肿瘤活性。结果显示原核表达载体构建成功,重组蛋白成功诱导表达,并初步证明了重组蛋白具有抑制肿瘤细胞增殖活性的功能,为后续抗肿瘤机制的探究奠定了基础。
