通过互补大肠杆菌发酵突变体克隆乳酸乳球菌adhE基因，该基因编码一种多功能乙醇脱氢酶。
Cloning of the Lactococcus lactis adhE gene, encoding a multifunctional alcohol dehydrogenase, by complementation of a fermentative mutant of Escherichia coli.
作者信息
Arnau J, Jørgensen F, Madsen S M, Vrang A, Israelsen H
机构信息
Biotechnological Institute, Hørsholm, Denmark.
出版信息
J Bacteriol. 1998 Jun;180(12):3049-55. doi: 10.1128/JB.180.12.3049-3055.1998.
Abstract
The Lactococcus lactis adhE gene, which encodes a multifunctional alcohol dehydrogenase, has been cloned and characterized. A DNA fragment encoding the putative alcohol dehydrogenase domain of the AdhE protein was cloned by screening an L. lactis genomic library in a fermentative mutant of Escherichia coli and selecting for the ability to grow anaerobically. Further analysis of the clone obtained allowed the cloning of the entire adhE gene sequence. Analysis of adhE expression in L. lactis during anaerobiosis showed induction at the transcriptional level, especially in medium containing glucose. Constructed mutant strains produced reduced amounts of ethanol under anaerobic conditions. With the L. lactis gene as a probe, adhE homologs were found in other industrially relevant lactic acid bacteria.
摘要
编码多功能乙醇脱氢酶的乳酸乳球菌adhE基因已被克隆并进行了特性分析。通过在大肠杆菌的发酵突变体中筛选乳酸乳球菌基因组文库，并选择厌氧生长能力，克隆了编码AdhE蛋白假定乙醇脱氢酶结构域的DNA片段。对所得克隆的进一步分析使得整个adhE基因序列得以克隆。对乳酸乳球菌在厌氧条件下adhE表达的分析表明，在转录水平上有诱导作用，尤其是在含有葡萄糖的培养基中。构建的突变菌株在厌氧条件下产生的乙醇量减少。以乳酸乳球菌基因作为探针，在其他与工业相关的乳酸菌中发现了adhE同源物。
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