摘要
目的:为了在枯草芽孢杆菌中整合表达极端耐热木聚糖酶。方法:将嗜热网球菌(Dictyoglomus thermophilum)Rt46B.1的极端耐热木聚糖酶基因xynB通过穿梭载体pDL整合到B.subtilis168染色体上,使其实现表达。结果:极端耐热木聚糖基因在枯草芽孢杆菌中成功整合并表达。结论:基因工程菌B.subtilis168-xynB能外泌表达极端耐热木聚糖酶,且表达水平为0.732IU/mL,比在大肠杆菌中的高。酶学性质表明,此酶分子量约为24kD,其最适反应温度为85℃,最适反应pH值为6.5,且在弱碱性条件下稳定。
Objective： In order to expressive the extreme thermophile xylanase gene xynB from Rt46B. 1 （Dictyoglomus thermophilum）. Method： The gene was cloned into the integration vectors of pDL and transform into host cell of B. subtilis 168. Result：The gene of xynB was recombinanted into chromatic body of B. subtilis 168 by double crossover homologous sequence substitution. Conclusion： The recombinant which has chlorampenieol resistance can express xylanase activity. Xylanase B（24kD） was successfully expressed （0. 7321U/mL） with higher - level production than expression system of E. eoli in LB. The recombinant protein of XynB expressed in B. subtilis 168 showed extreme thermostability and pH stability, which was optimally active at 85℃ and at pH 6.5. The recombinant XynB expressed in B. subtilis 168 is of great use in a variety of industrial and agrieuhural applications.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第1期15-18,共4页
Biotechnology
基金
新疆自治区高新技术项目(200515124)资助
关键词
极端耐热木聚糖酶基因
枯草芽孢杆菌
同源重组
双交换整合
thermostable xylanase gene
B. subtilis 168
homologous recombination
double crossover integration
