| 解淀粉芽孢杆菌中性植酸酶基因的克隆及其在毕赤酵母中的表达 |
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| 引用本文： | 张建云,崔树军,谷立坤,武秀琴. 解淀粉芽孢杆菌中性植酸酶基因的克隆及其在毕赤酵母中的表达[J]. 河南科技大学学报(自然科学版), 2010, 31(2): 69-72 |
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| 作者姓名： | 张建云 崔树军 谷立坤 武秀琴 |
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| 作者单位： | 1. 河南工程学院,资源与环境工程系,河南,郑州,451191
2. 河南工程学院,资源与环境工程系,河南,郑州,451191;中国科学院,研究生院,北京,100049 |
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| 基金项目： | 河南省科技攻关项目(092102210212);;河南工程学院青年基金项目(Y2007031) |
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| 摘 要： | 采用PCR法从解淀粉芽孢杆菌BA11中克隆到一个中性植酸酶phyc基因,将该基因克隆到毕赤酵母表达载体pPIC9K上并电转化至宿主细胞GS115后进行诱导表达。SDS-PAGE试验表明:该重组中性植酸酶在毕赤酵母宿主细胞中实现了高效分泌性表达。植酸酶活性测定结果显示阳性克隆子在诱导72h酶活性达到最高值,活性为2330U/L。
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| 关 键 词： | 中性植酸酶 毕赤酵母 解淀粉芽孢杆菌 异源表达 |
Cloning and Expression of Neutral Phytase Gene from Bacillus Amyloliquefaciens in Pichia Pastoris |
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| ZHANG Jian-Yun,CUI Shu-Jun,GU Li-Kun,WU Xiu-Qin. Cloning and Expression of Neutral Phytase Gene from Bacillus Amyloliquefaciens in Pichia Pastoris[J]. Journal of Henan University of Science & Technology:Natural Science, 2010, 31(2): 69-72 |
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| Authors: | ZHANG Jian-Yun CUI Shu-Jun GU Li-Kun WU Xiu-Qin |
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| Affiliation: | 1.College of Resource & Environment Science/a>;Henan Institute of Engineering/a>;Zhengzhou 451191/a>;China/a>;2.Graduate University of Chinese Academy of Sciences/a>;Beijing 100049/a>;China |
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| Abstract: | Phyc gene was amplified from Bacillus amyloliquefaciens genomic DNA by PCR.The Phyc gene was inserted into the expression vector pPIC9K and transformed into the Pichia pastoris strain GS115 by elect-roporation.Then the Phyc gene was expressed by induction of methyl alcohol.SDS-PAGE analysis showed that recombinant neutral phytase was successfully produced at a high level in Pichia pastoris.And after induced for 72 h,the enzyme activity can reach 2 330 U/L. |
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| Keywords: | Neutral phytase Pichia pastoris Bacillus amyloliquefaciens Heterologous expression |
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