提高内切葡聚糖酶活力及其在毕赤酵母中的表达研究  

Improvement of endoglucanase activity and its expression in Pichia pastoris

作　　者：唐自钟[1] 刘姗[1] 晋海军[1] 孙蓉[1] 陈惠[1] 韩学易[1] 

机构地区：[1]四川农业大学生命科学与理学院,四川雅安625014

出　　处：《扬州大学学报（农业与生命科学版）》2015年第1期72-77,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：四川省科技支撑计划项目(2008Z0150)

摘　　要：以中性内切葡聚糖酶基因EG和真菌Corticium rolfsii的碳水化合物结合模块（FCBM）为模块，构建融合基因重构体EG-FCBM和CD-FCBM，并利用高效表达载体在毕赤酵母中对其进行高效表达。酶活及性质分析显示，EG-FCBM和CD-FCBM在诱导表达84～96h后，其对微晶纤维素的活力分别为951和676U·mL^-1，较原始基因EG（526U·mL^-1）提高81％、28％，而酶学性质两者间无较大差异。这一结果表明，FCBM在催化水解纤维素的过程中有着极为重要的作用，通过增加FCBM来提高纤维素酶活性方法可行。Redesigned endoglucanases enhanced FCBM from the Corticiurn rolfsii, EG-FCBM and CD-FCBM respec- tively, were constructed in this study. The redesigned genes were expressed in P. pastoris, and their characters were also discussed. The enzymatic activities of EG, ECr-FCBM and CD-FCBM in Pichia pastoris cultivation supernatant reached 526, 951 and 676 U · mL-1 respectively. EG-FCBM and CD-FCBM showed 81% and 280% enzymatic activity increase compared to EG. The optimal pH, temperature, pH stability and heat stability between ECrFCBM and CD-FCBM had little difference. The results indicated that the FCBM from the Corticiurn rolfsii can improve EG＇s catalytic power.

关 键 词：内切葡聚糖酶 碳水化合物结合模块 融合基因 酶学性质 

分 类 号：Q789[生物学—分子生物学]