Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis |
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Authors: | Yi Cao Li Liao Xue-wei Xu Aharon Oren Ce Wang Xu-feng Zhu Min Wu |
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Institution: | (1) College of Life Sciences, Zhejiang University, Hangzhou, 310058, People’s Republic of China;(2) Institute of Life Sciences, and the Moshe Shilo Minerva Center for Marine Biogeochemistry, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel |
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Abstract: | Alcohol dehydrogenase (ADH; EC: 1.1.1.1) is a key enzyme in production and utilization of ethanol. In this study, the gene
encoding for ADH of the haloalkaliphilic archaeon Natronomonas pharaonis (NpADH), which has a 1,068-bp open reading frame that encodes a protein of 355 amino acids, was cloned into the pET28b vector
and was expressed in Escherichia coli. Then, NpADH was purified by Ni-NTA affinity chromatography. The recombinant enzyme showed a molecular mass of 41.3 kDa by SDS-PAGE.
The enzyme was haloalkaliphilic and thermophilic, being most active at 5 M NaCl or 4 M KCl and 70°C, respectively. The optimal
pH was 9.0. Zn2+ significantly inhibited activity. The K
m value for acetaldehyde was higher than that for ethanol. It was concluded that the physiological role of this enzyme is likely
the catalysis of the oxidation of ethanol to acetaldehyde. |
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Keywords: | Natronomonas pharaonis Haloalkaliphilic Alcohol dehydrogenase Molecular cloning |
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