高产纤维素酶木霉REMI突变株的构建与筛选  

Insertional Mutagenesis of Trichoderma through Restriction Enzyme Mediated Integration (REMI) for Improvement of Cellulase Activity

作　　者：胡仕凤[1] 唐俊[2] 高永东[2] 刘力行[2] 高必达[1] 陈捷[2] 

机构地区：[1]湖南农业大学生物安全技术学院,长沙410128 [2]上海交通大学农业与生物学院,上海200240

出　　处：《上海交通大学学报（农业科学版）》2007年第6期513-518,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：农业部948项目[2006-G54(A)];上海市科委中法合作项目(063407060)

摘　　要：利用限制性内切酶介导的基因整合技术(REMI)转化T.koningii CICC 40144,共获得23个稳定的突变株。PCR检测结果表明质粒pV2已成功导入突变株中。此外,从已获得的木霉REMI突变株(178个T.atroviride T23的突变株、64个T.koningii T30的突变株和23个T.koningii CICC 40144的突变株)中筛选具有高纤维素酶活性的突变株,其中明显高于出发菌的菌株10余株(TK-2R-3、TK-2R-9、TK-2R-11、TK-2R-14、T30-B3、T30-C7、T23-A2H等)。研究发现出发菌纤维素酶活越高,其突变株纤维素酶活提高幅度越小。筛选获得了1株高产纤维素酶木霉突变株TK-2R-14,其CX酶活达37.6 U·mL-1,比出发菌提高8.05%,FPA酶活达16.9 U·mL-1,比出发菌提高19.01%。Trichoderma koningii strain CICC40144 was transferred with linear pV2 DNA by restriction enzyme mediated integration, and 23 mutants were obtained and further identified by conventional PCR technique. Among 178 mutants from T. atroviride strain T23, 64 mutants from T. koningii strain T30 and 23 mutants from T. koningii strain CICC40144, over 10 mutants were screened for improved cellulase activity as compared with their parent strains. Moreover one mutant among all screened mutants, TK-2R-14, was found with the highest Cx（37.6 U ·mL^-1） and FPA （16.9 U·mL^-1） activities, which were increased by 8.05% and 19.07%, respectively.

关 键 词：木霉 REMI转化 纤维素酶 

分 类 号：Q933[生物学—微生物学]