| 酿酒酵母乙醛脱氢酶的克隆与表达 |
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| 引用本文： | 黄瑞,王璐,沈微,张晓梅,许赣荣. 酿酒酵母乙醛脱氢酶的克隆与表达[J]. 工业微生物, 2009, 39(1): 22-27 |
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| 作者姓名： | 黄瑞 王璐 沈微 张晓梅 许赣荣 |
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| 作者单位： | 江南大学生物工程学院,江苏无锡,214122;江南大学医药学院,江苏无锡,2141 |
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| 摘 要： | 利用PCR技术从酿酒酵母（Saccharomyces cerevisiae W303-1A）总DNA中扩增得到1．9kb乙醛脱氢酶编码基因aldh，将其连接到表达载体pEtac，得到重组载体pEtac—aldh，重组载体在大肠杆菌JM109中得到高效表达。对含有aldh的基因工程菌进行表达研究表明：该菌株在37℃下，以1．0mmol／LIPTG诱导5h酶活力达到22．8U，比酶活力为15．0U／mg蛋白，而对照菌株检测不到酶活力，并且该菌的耐乙醛浓度可达3．2g／L。
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| 关 键 词： | 乙醛脱氢酶 酿酒酵母 克隆与表达 |
Cloning and expression of acetaldehyde dehydrogenase of Saccharomyces cerevisiae W303-1A |
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| HUANG Rui,WANG Lu,SHEN Wei,ZHANG Xiao-mei,XU Gan-rong. Cloning and expression of acetaldehyde dehydrogenase of Saccharomyces cerevisiae W303-1A[J]. Industrial Microbiology, 2009, 39(1): 22-27 |
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| Authors: | HUANG Rui WANG Lu SHEN Wei ZHANG Xiao-mei XU Gan-rong |
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| Affiliation: | HUANG Rui , WANG Lu , SHEN Wei , ZHANG Xiao-mei, XU Gan-rong (1. School of Biotechnology, Jiangnan University, Wuxi 214122, China; 2. School of Medication, Jiangnan University, Wuxi 214122, China) |
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| Abstract: | An acetaldehyde dehydrogenase （ aldh ） gene about 1.9kb was amplified with PCR from the total DNA of Saccharomyces cerevisiae. Aldh gene was inserted into the expression vector pEtac and transformed into E. coli JM109, finally the recombinant （pEtac -aldh ） was obtained and the acetaldehyde dehydrogenase （ aldh ） was efficiently expressed. Induced by 1.0mmol/L IPTG for 5 hours, recombinant enzymes activity reached 22.8U at 37℃, the control enzymes activity was 15.0U/mg protein. However, enzyme activity of E. coli JM109 （pEtac） was not detectable yet. The recombinant JM109 grew well at the acetaldehyde concentration of 3.2g/L, obviously higher than that of the control. |
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| Keywords: | Saccharomyces cerevisiae acetaldehyde dehydrogenase cloning and expression |
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