| 黑曲霉糖化酶基因在酿酒酵母中的表达 |
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| 引用本文： | 钱鹏,汤斌.黑曲霉糖化酶基因在酿酒酵母中的表达[J].安徽机电学院学报,2012(2):1-3,7. |
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| 作者姓名： | 钱鹏 汤斌 |
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| 作者单位： | 安徽工程大学微生物发酵安徽省工程技术研究中心,安徽芜湖241000 |
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| 摘 要： | 从黑曲霉eDNA文库中筛选出糖化酶基因，并在酿酒酵母中进行表达．阳性克隆在发酵培养基中培养60h后，产生的糖化酶酶活力达到峰值为4．3U／mL．测定结果显示其糖化酶大小为1908bp，编码636个氨基酸残基组成的蛋白质．酶学性质分析显示该酶的最适反应温度为50℃，pH为5．0．经柱分离纯化其发酵上清液后，SDS-PAGE电泳方法，测得它的分子量大约为70kD，且条带清晰．
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| 关 键 词： | 黑曲霉 糖化酶 酿酒酵母 表达 |
Cloning and expression of glucoamylase genes from aspergilus niger cDNA library in S. cerevisiae |
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| QIAN Peng,TANG Bin.Cloning and expression of glucoamylase genes from aspergilus niger cDNA library in S. cerevisiae[J].Journal of Anhui Institute of Mechanical and Electrical Engineering,2012(2):1-3,7. |
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| Authors: | QIAN Peng TANG Bin |
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| Affiliation: | (Engineering Technology Research Center of Anhui Microbial Fermentation, Anhui Polytechnic University,Wuhu 241000,China) |
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| Abstract: | An expression cDNA library was constructed from aspergillus niger, then glucoamylase gene was isolated and efficiently expressed in S. cerevisiae. The results showed that the maximum activity of glucoamylase was 4.2 U/mL when cultivated for 60 h. Sequence analysis revealed that glucoamylase had 1 908 bp,which encodes a putative polypeptide of 636 amino acids. The enzyme proerties showed that the optimum pH and temperature was 5.0 and 50 ℃, the expressed protein was purified from the fermented supernatant using DEAE clumon and determined by SDS-PAGE. The results of SDS-PAGE also showed that the molecular weights of the enzyme was 70 kDa. |
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| Keywords: | aspergilus niger glucoamylase S cerevisiae expression |
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