白蚁纤维素酶结构域重组研究  

Recombinant cellulase of the NtEG

作　　者：刘慧琳[1] 曹以诚[1] 杨磊[1] 黄昱阳[1] 曾炳佳[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《广东农业科学》2011年第9期152-154,共3页Guangdong Agricultural Sciences

摘　　要：对来源于白蚁Nasutitermes takasagoensis的纤维素酶NtEG和来源于Thermomonospora fusca的耐热性纤维素酶E4进行同源建模和序列比较,利用重组PCR技术将E4的结合域与NtEG的催化域进行结构域重组,形成重组纤维素酶。并将得到的重组体置于毕氏酵母X33中表达并检测其催化效率。结果表明:重组纤维素酶的相对分子质量约为59 ku;用羧甲基纤维素法、滤纸法、微晶纤维素测得重组纤维素酶的活性分别为17.1、5.4、4.6 U/mL;重组纤维素酶可在毕赤酵母中成功表达。Cellulase NtEG,originated from termite Nasutitermes takasaoens and heat-resistant cellulase E4,originated from Thermomonospora fusca were compared sequences and modeled,recombined the binding domain of the E4 cellulase and the catalystic of the NtEG cellulase in structrure domain to get recombinant cellulase.The recombinant cellulases were expressed in pichia X33,and whose catalystic efficiency were checked.The results indicated that relative molecular mass was about 59 ku through SDS PAGE analysis,and recombinant yeast could generate transparent circle.The activity was 17.1 U/mL showed by carboxymethyl cellulose,and 5.4 U/mL showed by filter paper;the cellulase activity was 4.6 U/mL showed by microcrystalline cellulose.Recombinant cellulase expressed successfully in pichia.

关 键 词：重组纤维素酶 催化域 结合域 重组PCR 

分 类 号：Q556.2[生物学—生物化学]