系列木聚糖降解酶基因同向串连表达的研究  

Expression of Some Xylanolytic Enzyme Genes Tandem in the Same Direction

作　　者：刘思园[1] 邵蔚蓝[1] 裴建军[1] 吴华伟[1] 

机构地区：[1]南京师范大学微生物工程重点实验室,江苏南京210097

出　　处：《南京师大学报（自然科学版）》2006年第1期93-97,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30370034)

摘　　要：采用分子克隆操作方法,通过设计多个SD序列的多克隆位点,首次成功构建了一个木聚糖降解酶系列基因同向串连表达载体,经筛选鉴定获得具有极耐热性的α-阿拉伯呋喃糖苷酶基因(XarB)、葡萄糖醛酸酶基因(aguA)和木聚糖酶基因(XynB)同向串连的pHsh/XarB-aguA-XynB一株克隆菌.酶学分析表明重组菌株具有较好的阿拉伯糖苷酶、木糖苷酶、葡萄糖醛酸酶和木聚糖酶活性.酶活单位分别为:阿拉伯糖苷酶11.8 U/mL,β-木糖苷酶6.87 U/mL,α-葡萄糖醛酸酶1.53 U/mL,木聚糖酶6.58 U/mL.An expression vector is constructed with a tandem of three xylanolytic enzyme genes in the same direction, which includes arabinofuranosidase/xylosidase gene, α-glucuronidase gene, and xylanaseB gene. A new strategy is adopted to design three separate ribosome binding sites in the multiple cloning site of the expression vector. The analysis of enzyme activities of the engineered strain （ pHsh/XarB-αguA- XynB） show that it has 11.8 U/mL of arabinofuranosidase, 6.87 U/mL of β-xylosidase, 1.53 U/mL of α-glucuronidase and 6. 58 U/mL of xylanaseB activities.

关 键 词：表达载体 基因工程 同向串连 木聚糖降解酶 

分 类 号：Q814.1[生物学—生物工程]