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中温α-淀粉酶基因的克隆及在枯草芽孢杆菌中的高效表达
引用本文:刘逸寒,路福平,王建玲,胡博. 中温α-淀粉酶基因的克隆及在枯草芽孢杆菌中的高效表达[J]. 天津科技大学学报, 2011, 0(4): 1-5
作者姓名:刘逸寒  路福平  王建玲  胡博
作者单位:工业发酵微生物教育部重点实验室天津科技大学生物工程学院;
基金项目: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)
摘    要:利用高效表达栽体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AM...

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

High-Level Expression of the Medium-Temperature Alpha Amylase in Bacillus subtilis
LIU Yi-han,LU Fu-ping,WANG Jian-ling,HU Bo. High-Level Expression of the Medium-Temperature Alpha Amylase in Bacillus subtilis[J]. Journal of Tianjin University of Science & Technology, 2011, 0(4): 1-5
Authors:LIU Yi-han  LU Fu-ping  WANG Jian-ling  HU Bo
Affiliation:LIU Yi-han,LU Fu-ping,WANG Jian-ling,HU Bo(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science & Technology,Tianjin 300457,China)
Abstract: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 purification.The recombinant enzyme AMY had a molecular mass of 4.8×104.The optimum pH and temperature of AMY was 6.0 and 60 ℃.
Keywords:Bacillus subtilis  medium-temperature alpha amylase  high-level expression  purification  enzyme properties  
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