Cloning and Characterization of Ethanol-Regulated Esterase Genes in Acetobacter pasteurianus
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概要
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The esterase encoding genes, est1 and est2, were cloned from Acetobacter pasteurianus. Nucleotide sequence analysis of est1 revealed a gene of 954 bp, and est1 coded for an arylesterase with a molecular weight of 34863 Da consisting of 317 amino acids. The est2 gene contained an open reading frame composed of 1221 bp encoding an esterase with a molecular weight of 43389 Da consisting of 406 amino acids. The est1 gene showed some similarity, but the est2 gene showed no significant homology to other esterases reported in various microorganisms. Northern blot analysis of total RNA from A.pasteurianus revealed that transcription of the est1 gene was induced only when the cells were grown in a medium containing ethanol, and suggested that the est1 transcript is monocistronic. In contrast, transcription of the est2 gene was repressed in the presence of ethanol. In the absence of ethanol, expression of the est2-mRNA, capable of encoding a multiple number of proteins, was revealed by Northern blot analysis. In addition, deletion analysis indicated that the 5′-region of the est2 gene contained a cis-acting domain for est2 transcriptional regulation. Analysis of the est1 promoter using the chloramphenicol acetyltransferase gene as a reporter gene showed that the promoter within the 305-bp fragment upstream of the ATG initiation codon was responsible for the transcription in cells grown in the presence of ethanol. Primer extension analysis of est1-mRNA showed that the transcription initiation site was 49 bp upstream from the ATG initiation codon. The results of a gel mobility shift assay indicated that there is a regulatory protein related to est1 regulation, which may have some relation to the ethanol resistance of Acetobacter sp.
- 社団法人日本生物工学会の論文
- 1999-01-25
著者
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Koizumi Yukimichi
Department of Fermentation Science, Faculty of Applied Bio-Science, Tokyo University of Agriculture
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NAKANO Takayuki
Department of Electrical and Electronic Engineering, Niigata University
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Toda K
Faculty Of Engineering Niigata Institute Of Technology
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Kawamura Yukio
Department Of Fermentation Science Faculty Of Applied Bio-science Tokyo University Of Agriculture
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Kawamura Y
Akita Res. Inst. Food And Brewing (arif) Akita
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Kawamura Y
National Food Res. Inst. Ibaraki Jpn
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UDAKA SHIGEZO
Department of Fermentation Sciences, Tokyo University of Agriculture
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Kashima Y
National Inst. Of Advanced Industrial Sci. And Technol. (aist Kansai) Osaka Jpn
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Koizumi Y
Department Of Fermentation Science Faculty Of Applied Bio-science Tokyo University Of Agriculture
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KASHIMA Yasuhiro
Department of Fermentation Science, Tokyo University of Agriculture
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YANAGIDA Fujiharu
Department of Fermentation Science, Tokyo University of Agriculture
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NAKAJIMA Yusuke
Department of Fermentation Science, Faculty of Applied Bio-Science, Tokyo University of Agriculture
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TAYAMA Kenji
Central Research Institute, Nakano Vinegar Co., Ltd.
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Tayama K
Mitsukan Group Corp. Aichi Jpn
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Tayama Kenji
Central Research Institute Mitsukan Group Corporation
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Tayama Kenji
Nakano Biochemical Research Institute Nakano Vinegar Co. Ltd.
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Nakajima Yusuke
Department Of Fermentation Science Faculty Of Applied Bio-science Tokyo University Of Agriculture
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Kashima Yasuhiro
Department Of Chemistry Oklahoma State University
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Yanagida Fujiharu
Department Of Fermentation Science Faculty Of Applied Bioscience Tokyo University Of Agriculture
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Udaka S
Faculty Of Applied Bio-science Tokyo University Of Agriculture
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Udaka Shigezo
Department Of Brewing And Fermentation Tokyo University Of Agriculture
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Nakano Takayuki
Department Of Biosciences Teikyo University
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Koizumi Yukimichi
Department Of Fermentation Science Faculty Of Applied Bio-science Tokyo University Of Agriculture
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Koizumi Y
Kyoto Univ. Kyoto Jpn
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Udaka Shigezo
Dept. Of Food Science And Technology Faculty Of Agriculture Nagoya University
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Nakano Takayuki
Department Of Fermentation Science Tokyo University Of Agriculture
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YANAGIDA Fujiharu
Department of Brewing and Fermentation, Tokyo University of Agriculture
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