| 麦芽糖α-淀粉酶基因在枯草芽孢杆菌中的分泌表达 |
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| 引用本文： | 沈微,俞玲,陈献忠,樊游,王正祥.麦芽糖α-淀粉酶基因在枯草芽孢杆菌中的分泌表达[J].食品与发酵工业,2011,37(6). |
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| 作者姓名： | 沈微 俞玲 陈献忠 樊游 王正祥 |
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| 作者单位： | 江南大学生物工程学院工业生物技术教育部重点实验室,江苏无锡,214122 |
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| 基金项目： | 国家863高技术研究发展计划(2006AA10Z307) |
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| 摘 要： | 采用重叠PCR方法在麦芽糖α-淀粉酶编码基因5’端添加地衣芽孢杆菌α-淀粉酶基因信号肽编码区,获得重组基因BlMa。重组基因与芽孢杆菌表达载体pHY-P43连接后直接转化枯草芽孢杆菌,获得重组质粒pHY-P43-BlMa。枯草芽孢杆菌淀粉酶基因缺陷株1A717被用作BlMa基因表达宿主菌,重组菌命名为Bacillus subtilis/pHY-P43-BlMa。酶活检测和SDS-PAGE电泳均显示,B.subtilis/pHY-P43-BlMa表达的重组麦芽糖淀粉酶(BlMa)全部分泌到培养液中。HPLC检测表明,BlMa催化可溶性淀粉水解产物主要为麦芽糖。对B.subtilis/pHY-P43-BlMa摇瓶发酵条件进行优化。获得优化发酵培养基配方:10%玉米淀粉,2.5%药媒,0.3%(NH4)2SO4,0.03%CaCl2,0.1%NaH2PO4,在优化条件下重组菌发酵酶活为5.9 U/mL。
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| 关 键 词： | 麦芽糖α-淀粉酶 分泌表达 枯草芽孢杆菌 |
Secreted Expression of the Maltogenic Amylase in Bacillus subtilis |
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| Shen Wei,Yu Ling,Chen Xian-zhong,Fan You,Wang Zheng-xiang.Secreted Expression of the Maltogenic Amylase in Bacillus subtilis[J].Food and Fermentation Industries,2011,37(6). |
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| Authors: | Shen Wei Yu Ling Chen Xian-zhong Fan You Wang Zheng-xiang |
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| Affiliation: | Shen Wei,Yu Ling,Chen Xian-zhong,Fan You,Wang Zheng-xiang(The Key Laboratory of Industrial Biotechnology,Ministry of Education and School of Biotechnology,Jiangnan University,Wuxi 214122,China) |
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| Abstract: | The gene encoding maltogenic amylase was fused with signal peptide gene encoding amylase of Bacillus licheniformis by overlapping PCR.The fused gene(BlMa) was ligated with an expression vector of Bacillus,pHY-P43.B1Ma transforms Bacillus subtilis directly and results in the recombinant plasmid pHY-P43-BlMa.Bacillus subtilis 1A717 which is deficient in amylase was selected as expression host.The recombinant strain was designated as Bacillus subtilis / pHY-P43-BlMa.Enzyme activity assay and SDS-PAGE showed th... |
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| Keywords: | maltogenic amylase secreted expression Bacillus subtilis |
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