后熟期短的酿酒酵母工程菌构建

食品科学 ›› 2006, Vol. 27 ›› Issue (8): 83-86.

后熟期短的酿酒酵母工程菌构建

刘增然, 张光一, 鞠国泉, 冉会来, 蔡亚男

河北经贸大学生物科学与工程学院

出版日期:2006-08-15 发布日期:2011-10-14

Construction of A Strain of Saccharomyces cerevisiae with Less Maturation Time

LIU Zeng-Ran, ZHANG Guang-Yi, JU Guo-Quan, RAN Hui-Lai, CAI Ya-Nan

College of Bioscience and Bioengineering,Hebei Economic and Trade University,Shijiazhuang 050061,China

Online:2006-08-15 Published:2011-10-14

摘要/Abstract

摘要： 借助质粒pBluescriptM13-,以淀粉酶基因(AMY)为筛选标记构建乙酰乳酸合成酶基因(ILV2)的重组整合质粒pMGI。用重组质粒pMGI所含ilv2∷AMY嵌合基因片段转化工业酿酒酵母菌Sc11,获得转化子(Sc11-ilv)。Sc-11-ilv不含细菌载体序列和酵母抗药性标记,所表达的乙酰乳酸合成酶活性降低30%;模拟发酵实验表明Sc11-ilv发酵液的双乙酰含量降低70%,Sc11-ilv发酵性能保持不变,遗传稳定性为100%,可以比较安全地用于啤酒生产。

关键词: 工业酿酒酵母菌, &alpha, -乙酰乳酸合成酶基因(ILV2), 基因破坏, 双乙酰, 发酵

Abstract: An integrative plasmid pMGI carrying yeast α-acetolactate synthase gene(ILV2),that used the cloned amylase gene(AMY)as the selective marker,was constructed from pBluescript M13-.The ilv2∷AM expression cassette released from pMGI was transformed into an industrial strain of Saccharomyces cerevisiae and integrated at the ILV2 gene of the host genome through one-step replacement,gaining transformants(Sc11-ilv)free of undesired sequences(bacterial or viral vector sequences and yeast selective marker).The α-acetolactate synthase activity of the recombinant strain was lowered by 30%.Fermentation tests confirmed that the diacetyl concentration was reduced by 70% in fermented wort by the recombinant strain,while the brewing performances of the recombinant strain were retained.This type of integration is genetically stable even after 100 generations of cell multiplication under non-selective conditions and can be used in beer production safely.

Key words: an industrial strain of S.cerevisiae, acetolactate synthase gene(ILV2), gene disruption, diacetyl, fermentation

刘增然, 张光一, 鞠国泉, 冉会来, 蔡亚男. 后熟期短的酿酒酵母工程菌构建[J]. 食品科学, 2006, 27(8): 83-86.

LIU Zeng-Ran, ZHANG Guang-Yi, JU Guo-Quan, RAN Hui-Lai, CAI Ya-Nan. Construction of A Strain of Saccharomyces cerevisiae with Less Maturation Time[J]. FOOD SCIENCE, 2006, 27(8): 83-86.

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