不同连接肽在硫酸软骨素酶ABC I重组表达中的应用研究  

Application of different linker peptides in Chondroitinase ABC I recombinant expression

作　　者：李晔[1] 陈振娅 王瑞丽 沈荣 危晴[1] 陈亮[1] LI Ye;CHEN Zhenya;WANG Ruili;SHEN Rong;WEI Qing;CHEN Liang(College of Biotechnology,Beijing Polytechnic,Beijing 100176,China;College of Life Science,Beijing Institute of Technology,Beijing 100081,China)

机构地区：[1]北京电子科技职业学院生物工程学院,北京100176 [2]北京理工大学生命学院,北京100081

出　　处：《中国酿造》2020年第9期121-125,共5页China Brewing

基　　金：北京市自然基金面上项目(2182019);北京市优秀人才培养资助(拔尖自然科学)(2020Z002-002-KWT);北京电子科技职业学院院内科技类重点课题(2019Z002-033-KXZ)。

摘　　要：硫酸软骨素酶ABC I(ChSase ABC I)是一类能够将硫酸软骨素、软骨素、透明质酸等糖胺多糖降解为寡糖及不饱和二糖的裂解酶。该研究将硫酸软骨素酶ABC I基因与麦芽糖结合蛋白(MBP)基因分别用FFFFF和RRRRR两种连接肽连接,并克隆到pMAL-c2X载体上,在大肠杆菌(Escherichia coli)BL21(DE3)高效表达。结果表明,重组酶MBP-FFFFF-ChSase ABC I的酶活和比酶活分别为716.5 IU/L发酵液和1.15 IU/mg蛋白,重组酶MBP-RRRRR-ChSase ABC I的酶活和比酶活分别为741.0 IU/L发酵液和1.13 IU/mg蛋白。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结果表明,重组酶MBP-FFFFF-ChSase ABC I和MBP-RRRRR-ChSase ABC I的分子质量均为130 kDa,并且都为可溶性蛋白。Chondroitinase ABC I(ChSase ABC I)is a kind of lyases that can degrade glycosaminoglycans such as chondroitin sulfate,chondroitin and hyaluronic acid into oligosaccharides and unsaturated disaccharides.The ChSase ABC I gene was linked to maltose binging protein(MBP)gene using two linker peptides including FFFFF and RRRRR,respectively,cloned into pMAL-c2X vector,and successfully expressed in Escherichia coli BL21(DE3).The results showed that the enzymatic activity and specific enzyme activity of the recombinase MBP-FFFFF-ChSase ABC I were 716.5 IU/L fermentation broth and 1.15 IU/mg protein,respectively,and the recombinase MBP-RRRRR-ChSase ABC I were 741.0 IU/L fermentation broth and 1.13 IU/mg protein,respectively.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)results showed that the molecular weights of the recombinases MBP-FFFFF-ChSase ABC I and MBP-RRRRR-ChSase ABC I were 130 kDa,and they were soluble proteins.

关 键 词：硫酸软骨素酶ABC I 普通变形杆菌 连接肽 

分 类 号：Q815[生物学—生物工程]