重组丙酮丁醇梭菌的构建及其木质纤维素ABE发酵  

ABE fermention from cellulose by recombinant Clostridium acetobutylicum

作　　者：林丽华[1,2] 刘春宇[1] 郭媛[2] 汤宏赤 庞浩[1,2] Lihua Lin;Chunyu Liu;Yuan Guo;Hongchi Tang;Hao Pang(College of Life Science and Technology,Guangxi University,Nanning 530004;National Engineering Research Center for Non - Food Biorefinery,State Key Laboratory of Non - Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio - refinery,Guangxi Academy of Sciences,Nanning 530007,China)

机构地区：[1]广西大学生命科学与技术学院,广西南宁530004 [2]广西科学院,国家非粮生物质能源工程技术研究中心,非粮生物质酶解国家重点实验室,广西生物炼制重点实验室,广西南宁530007

出　　处：《生物技术》2018年第6期592-596,共5页Biotechnology

基　　金：国家自然科学基金项目(“纤维寡糖转运系统在丙酮丁醇梭菌中的整合及其对纤维寡糖转运和利用的影响研究”,No.21366007);广西科技计划项目(“南地区种养殖业面源污染控制及生物质能源综合利用关键技术研究”,No.桂科AD16380016);广西自然科学基金项目(“cbpC蛋白在有机溶剂中的稳定性及其对纤维糊精跨膜转运的影响”,No.2015GXNSFBA139084;“astE基因缺失提高大肠杆菌丁醇耐受能力的分子机制研究”,No.2015GXNSFBA139048)

摘　　要：[目的]提高丙酮丁醇梭菌降解利用纤维素原料的能力.[方法]将甲基化的重组表达载体pSOS95-cel9 电转化至丙酮丁醇梭菌ATCC824.[结果]成功构建重组菌株ATCC824 /pSOS95-cel9.通过荧光定量PCR 检验到外源纤维素酶基因cel9 在重组丙丁梭菌中的转录,24 h 的相对表达量是12 h 的27.1 倍.重组菌ATCC824 /pSOS95-cel9 发酵滤纸产丙酮0.05 g /L、丁醇0.08 g /L、乙醇0.71 g /L,发酵蔗渣水解液产0.78 g /L、1.09 g /L、0.97 g /L,各溶剂产量均显著高于空质粒对照菌株.[结论]重组菌能利用滤纸及蔗渣水解液进行ABE 发酵,外源纤维素酶基因cel9 的重组表达提高了工程菌株利用纤维素原料的能力.[Objective]To improve the ability of Clostridium acetobutylicum to utilize cellulose-containing raw materials.[Methods]Methylated recombinant expression vector pSOS95-cel9 was transformed into the Clostridium acetobutylicumATCC824 by electrotransformation.[Results] The engineering strain ATCC824 /pSOS95-cel9 was constructed successfully.The results of real-time quantitative PCR demonstrated that the cellulase gene cel9 was transcribed successfully in recombinantClostridium acetobutylicum,the relative quantification of cel9 gene expression of 24 hours is 27.1 times that of 12 hours.For fermentation of filter paper ,the recombinant strain ATCC824/pSOS95-cel9 produced 0.05 g /L of acetone,0.08 g /L ofbutanol,and 0.71 g /L of ethanol.For fermentation of sugarcane bagasse hydrolyzate,the yield of recombinant strain were 0.78g /L,1 .09 g /L,0.97 g /L,respectively.Each solvent yield was significantly higher than that of the empty plasmid control strain.[Conclusion]Recombinant strain can effectively ferment the filter paper and bagasse hydrolysate to Acetone-Butanol-Ethanol,The introduction of cellulase gene cel9 significantly improved the ability of Clostridium acetobutylicum to utilize cellulose -containing raw materials.

关 键 词：纤维素酶 重组丙酮丁醇梭菌 滤纸 甘蔗渣 ABE 发酵 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]