纤维素酶EGA基因在枯草芽孢杆菌中的表达及其产物性质研究  

Research on Expression of Cellulase EGA Gene in Bacillus Subtilis and Properties of Its Product

作　　者：张漫莉[1] 耿新伟[1] 王梦婷[1] 蒋磊[1] 赵辅昆[1] 陈玮[1] 

机构地区：[1]浙江理工大学蛋白质组学与分子酶学实验室,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2013年第3期389-393,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：浙江省大学生科技成果推广项目(2011R406051)

摘　　要：枯草芽孢杆菌Bacillus subtilis是一种高效的外源蛋白表达菌株,能将目的蛋白分泌到细胞外的培养基中。从福寿螺胃液共生菌株Bacillus sp.Strain AC-1基因组中克隆纤维素酶EGA的基因序列,利用基因工程技术构建纤维素酶EGA的重组枯草芽孢杆菌表达载体pAUsp-ega。该载体含有一个强启动子和一段信号肽,经淀粉诱导能表达外源蛋白。以枯草芽孢杆菌蛋白酶缺失型菌株WB700为宿主菌,表达得到的重组纤维素酶EGA以羧甲基纤维素钠(CMC-Na)为底物得到的培养基上清酶活力达到1 120U/L。该纤维素酶在pH 6.0表现最大水解活力,最适反应温度为60℃,在酸性条件下稳定性良好,具有较好的应用前景。Bacillus subtilis is an efficient exogenous protein expression stain which can secrete the target protein into extracellular culture medium. This research clones the gene order of cellulase EGAA from ampullaria gigas gastric juice symbiotic strain Bacillus sp. Strain AC-1 genome and establishes expression vector pAUsp-ega of recombinant bacillus subtilis of cellulase ega using genetic engineering technology. This vector contains a strong promotor and signal peptide and can express exogenous protein with induction by amylum. The enzyme activity of culture medium supernatant obtained by recombinant cellulase ega with CMC-Na as substrate and bacillus subtilis protease deletion form stain WB700 as host bacteria reaches 1 120 U/L. This eellulase shows the maximum hydrolysis vigor when pH=6.0; its most appropriate reaction temperature is 60℃. It has a good stability under acidic condition and has a good application prospect.

关 键 词：枯草芽孢杆菌 纤维素酶EGA分泌 羧甲基纤维素钠 

分 类 号：Q786[生物学—分子生物学]