TY - JOUR
T1 - Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis
AU - Cao, Yi
AU - Liao, Li
AU - Xu, Xue Wei
AU - Oren, Aharon
AU - Wang, Ce
AU - Zhu, Xu Feng
AU - Wu, Min
PY - 2008/5
Y1 - 2008/5
N2 - 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 Km 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.
AB - 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 Km 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.
KW - Alcohol dehydrogenase
KW - Haloalkaliphilic
KW - Molecular cloning
KW - Natronomonas pharaonis
UR - https://www.scopus.com/pages/publications/43049165328
U2 - 10.1007/s00792-007-0133-7
DO - 10.1007/s00792-007-0133-7
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C2 - 18189118
AN - SCOPUS:43049165328
SN - 1431-0651
VL - 12
SP - 471
EP - 476
JO - Extremophiles
JF - Extremophiles
IS - 3
ER -