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| 耐热型α-葡萄糖苷酶基因在大肠杆菌中的克隆及表达 | |
| 引用本文： | 周勤华,牟航,周盛梅.耐热型α-葡萄糖苷酶基因在大肠杆菌中的克隆及表达[J].工业微生物,2011,41(6):67-71. |
| 作者姓名： | 周勤华 牟航 周盛梅 |
| 作者单位： | 1. 嘉兴学院医学院,浙江嘉兴,314001 2. 浙江清华长三角研究院,浙江嘉兴,314001 3. 嘉兴学院生物与化学工程学院,浙江嘉兴,314001 |
| 基金项目： | 嘉兴市科技局科技研究计划项目 |
| 摘 要： | 应用PCR法从火山口嗜热菌中克隆出了α-葡萄糖苷酶基因，并将该基因导入大肠杆菌，获得了稳定表达的大肠杆菌菌株。同时对其表达产物进行了酶学性质分析。结果表明：该基因长1587bp，编码501个氨基酸，为新舡葡萄糖苷酶同源基因，已将该序列在Genbank中登记；其表达产物经SDS-PAGE分析表明，相对分子质量约为66kD；该α-葡萄糖苷酶的最适温度为90℃，最适pH值为7．0，在80℃放置3h，酶活仍能达到94％以上。结果表明该酶具有优良的耐热性，将有广泛的工业应用前景。 |
| 关 键 词： | α-葡萄糖苷酶 嗜热菌 PCR 耐热性 |
Cloning and expression of heat resistant α-glucosidase from Thermophilic bacilli and its biochemical characterization |
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| ZHOU Qin-hua,MU Hang,ZHOU Sheng-mei.Cloning and expression of heat resistant α-glucosidase from Thermophilic bacilli and its biochemical characterization[J].Industrial Microbiology,2011,41(6):67-71. | |
| Authors: | ZHOU Qin-hua MU Hang ZHOU Sheng-mei |
| Institution: | 1. College of Medicine, Jiaxing University, Jiaxing, Zhejiang 314001; 2. Yangtze Delta Region Institute of Tsinghua University, Jiaxing, Zhejiang 314001 ; 3. College of Biology and Chemical Engineering, Jiaxing University, Jiaxing, Zhejiang 314001) |
| Abstract: | Using PCR method, α-glucosidase gene consisted of 1587 nucleotide and encoded 501 amino acids was cloned from Thermophilic bacilli and expressed in E. coli BL21 system. The subsequent characterization elucidated that α-glucosidase was extracted from BL21 bacteria with a molecular mass of 66 kD and it had optimum activity at 900C and pH 7.0. Interestingly, the relative activity was 94% when incubated at 80℃ for 3 h, suggesting that it could meet the industrial demands due to the properties of extreme thermal activity and stability. |
| Keywords: | α-glucosidase thermophiles polymerase chain reaction heat resistant |
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