瑞氏木霉内切-β-1,4葡聚糖酶基因Egl1的分子改造  

Molecular Modification of Endo-β-1,4-glucanase Gene Egl1 from Trichoderma reesei

作　　者：陈小玲[1] 陈东[1] 芦志龙[1] 

机构地区：[1]广西科学院非粮生物质酶解国家重点实验室国家非粮生物质能源工程技术研究中心广西生物炼制重点实验室,广西南宁530007

出　　处：《广西科学》2011年第3期264-268,共5页Guangxi Sciences

基　　金：广西科技攻关项目(桂科转10123005-17);广西科学院基本科研业务费项目(10YJ25SW12)资助

摘　　要：为了提高瑞氏木霉（Trichoderma reesei）的纤维素酶活力,用分子改造的方法改造其内切-β-1,4葡聚糖酶基因Egl1。用DNaseⅠ消化Egl1,回收50~200bp的片段,用T4 DNA连接酶连接回收的片段,进行PCR反应,将PCR产物转入瑞氏木霉原生质体,用比较滤纸酶活力的方法筛选纤维素酶活力提高的菌株。结果筛选到2株滤纸酶活力比出发菌株提高的菌株Egl36-4和Egl33-4B2Z2,其酶活力分别比出发菌株提高了2.8倍和3.6倍。Endo-β-1,4-glucanase gene Egl1 was digested with DNase Ⅰ,fragments of 50~200bp were collected and randomly linked with T4 DNA,then were used as templates for sequences amplification by polymerase chain reaction（PCR）.The shuffling sequences were transformed to T.reesei by protoplast transformation.Two mutants with higher cellulase activity were selected.The filter paper activity（FPA） of two mutants named Egl36-4 and Egl33-4B2Z2 were 2.8 and 3.6 times higher than that of original strain,respectively.

关 键 词：瑞氏木霉 纤维素酶 分子改造 

分 类 号：Q939.5[生物学—微生物学] Q78