摘要
利用高效表达载体pWB980,实现了Bacillus subtilis BF7658中温α–淀粉酶基因amy在B.subtilis DB403中的高效表达,活力达到770,U/mL.经多步纯化,重组酶AMY的比活达到35.8,U/mg,纯化倍数为1.7,获得凝胶电泳条带单一的蛋白样品,经SDS-PAGE检测,重组酶AMY相对分子质量为4.8×104.对酶学性质进行分析,重组酶AMY的最适反应温度为60,℃,最适反应pH为6.0.
The medium-temperature alpha amylase gene from Bacillus subtilis BF768 was cloned into vector pWB980 and high-level expressed in B. subtilis DB403. The activity of AMY in the supernatant of the culture medium reached a maximum of 770 U/mL. By multi-step purification, the specific activity of AMY was up to 35.8 U/mg with a 1.7-fold purifica- tion. The recombinant enzyme AMY had a molecular mass of 4.8 ×10^4. The optimum pH and temperature of AMY was 6.0 and 60 ℃.
出处
《天津科技大学学报》
CAS
2011年第4期1-5,共5页
Journal of Tianjin University of Science & Technology
基金
天津市科技攻关重点培育项目(06YFGPSH03500)
天津科技大学自然科学基金资助项目(20090205)
关键词
枯草芽孢杆菌
中温α-淀粉酶
高效表达
纯化
酶学性质
Bacillus subtilis
medium-temperature alpha amylase
high-level expression
purification
enzyme properties
