| 绿色糖单孢菌麦芽糖α-淀粉酶基因在枯草芽孢杆菌中的高效分泌表达 |
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| 引用本文： | 柳梅梅,谢敏,杨燕芳,刘滔滔,杜丽琴,梁智群,韦宇拓.绿色糖单孢菌麦芽糖α-淀粉酶基因在枯草芽孢杆菌中的高效分泌表达[J].广西科学,2016,23(1):12-18. |
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| 作者姓名： | 柳梅梅 谢敏 杨燕芳 刘滔滔 杜丽琴 梁智群 韦宇拓 |
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| 作者单位： | 广西大学生命科学与技术学院,广西南宁530005;亚热带农业生物资源保护与利用国家重点实验室,广西南宁 530005 |
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| 基金项目： | 国家自然科学基金项目(31160311)，广西自然科学基金项目(2012GXNSFAA053051) |
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| 摘 要： | 【目的】构建高产麦芽糖α-淀粉酶的工程菌株并实现高效分泌表达。【方法】PCR扩增麦芽糖α-淀粉酶基因sva,再与大肠杆菌-枯草芽孢杆菌麦芽糖诱导型穿梭载体pHCMCO4-Pglv连接,构建重组质粒pHCMCO4-Pglv-sva并转入枯草芽孢杆菌进行表达,对重组酶进行SDS-PAGE分析,然后对重组菌株的生长及发酵条件进行优化。【结果】成功构建重组质粒pHCMCO4-Pglv-sva并在枯草芽孢杆菌中实现分泌表达,SDS-PAGE分析发现,在55kDa处得到特异性蛋白条带。单因素试验结果显示,重组菌的最适诱导温度为35℃,最适接种量为4%(V/V),最适装液量为30mL。正交实验结果显示,重组菌的最佳发酵条件组合是诱导温度37℃、接种量5%(V/V)、装液量25mL,在此条件下发酵液粗酶活力达到257.3698U/mL。装液量对重组菌的产酶量影响显著。【结论】成功构建能高效分泌表达麦芽糖α-淀粉酶的枯草芽孢杆菌工程菌株,经发酵条件优化后,重组酶产量显著提高10倍左右。
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| 关 键 词： | 麦芽糖a-淀粉酶 枯草芽孢杆菌 高效表达 优化 |
| 收稿时间： | 2015/11/16 0:00:00 |
| 修稿时间： | 2016/2/28 0:00:00 |
High Level Secretion Expression of Maltogenic α-amylase from Saccharomonospora viridis in Bacillus subtilis |
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| LIU Meimei,XIE Min,YANG Yanfang,LIU Taotao,DU Liqin,LIANG Zhiqun and WEI Yutuo.High Level Secretion Expression of Maltogenic α-amylase from Saccharomonospora viridis in Bacillus subtilis[J].Guangxi Sciences,2016,23(1):12-18. |
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| Authors: | LIU Meimei XIE Min YANG Yanfang LIU Taotao DU Liqin LIANG Zhiqun and WEI Yutuo |
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| Institution: | College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China and College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilizaiton of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China |
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| Abstract: | |
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| Keywords: | maltose a-amylase Bacillus subtilis high efficient expression optimization |
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