| 枯草芽孢杆菌克隆与表达微小毛霉凝乳酶基因 |
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| 引用本文： | 李自刚,李凤娟,云丽,吴坤,赵跃进,屈凌波.枯草芽孢杆菌克隆与表达微小毛霉凝乳酶基因[J].中国酿造,2012(10):68-72. |
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| 作者姓名： | 李自刚 李凤娟 云丽 吴坤 赵跃进 屈凌波 |
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| 作者单位： | [1]郑州牧业工程高等专科学校,河南郑州450011 [2]河南农业大学,河南郑州450001 [3]三门峡龙飞生物工程有限公司,河南三门峡472100 [4]河南工业大学,河南郑州450001 |
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| 基金项目： | 河南省重大公益科研招标项目资金资助（HNZB[2008]N84） |
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| 摘 要： | 从自制的酒酿利用酪蛋白培养基分离纯化到了一株产凝乳酶的微小毛霉菌株（ZZMZ-19），从ZZM-19菌株的cDNA文库筛选到了两个凝乳酶基因chl和ch2并实现了凝乳酶基因ch1和ch2在枯草芽孢杆菌菌株（ZZMZ-01）中的的克隆与表达。chl和曲2阳性克隆菌株在酪蛋白培养基r1]发酵30h左右的时间凝乳酶酶活达到最人，分别为48．27SU／mL和41．02SU／mL，相比出发菌株发酵时间缩短了15h。用交联葡聚糖凝胶柱G100分离纯化其发酵卜清液后，用十二烷基硫酸钠聚丙烯酰胺凝胶电泳方法测得CHII和cHIII分子草分别为32ku和30ku。
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| 关 键 词： | 酒酿 微小毛霉 凝乳酶 枯草芽孢杆菌 克隆 表达 |
Cloning and expression of chymosin gene from Mucor pusillus cDNA library in Bacillus subtilis |
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| LI Zigang,LI Fengjuan,LI Yun,WU Kun,ZHAO Yuejin,QU Lingbo.Cloning and expression of chymosin gene from Mucor pusillus cDNA library in Bacillus subtilis[J].China Brewing,2012(10):68-72. |
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| Authors: | LI Zigang LI Fengjuan LI Yun WU Kun ZHAO Yuejin QU Lingbo |
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| Affiliation: | 1.Zhengzhou College of Animal Husbandry Engineering, Zhengzhou 450011, China; 2.College of Life Science of Henan Agricultural University, Zhengzhou 450002, China, 3.Sanmenxia Longfei Biological Engineering Co., Ltd., Zhengzhou 472100, China; 4. Henan University of Technology, Zhengzhou 450011, China) |
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| Abstract: | A high chymosinproducing Mucorpusillus strains （ZZMZ-19） was isolated from home-made Jiu-Niang using casein medium, and two novel chymosin genes chl and ch2 were cloned from cDNA library of ZZMZ-19 and expressed in Bacillus subtilis strains （ZZMZ-01）. The recombined ZZMZ-01 strain was cultured in the casein medium. The results showed that the maximum activities of recombined chymosin were obtained after 30h fermentation, which was 15h earlier than that of the original strain Mucor pusillus ZZMZ-19. The values were 48.27SU/ml and 41.02SU/ml, respec- tively. The expressed chymosin were purified using cross-linking glucan gel column and determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was found that the molecular weights of CH1 and CH2 enzymes were 32 ku and 30 ku, respectively. |
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| Keywords: | Jiu-Niang Mucorpusillus chymosin Bacillus subtilis clone expression |
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