Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis

Yi Cao; Li Liao; Xue Wei Xu; Aharon Oren; Ce Wang; Xu Feng Zhu; Min Wu

doi:10.1007/s00792-007-0133-7

Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis

Yi Cao, Li Liao, Xue Wei Xu, Aharon Oren, Ce Wang, Xu Feng Zhu, Min Wu^*

^*Corresponding author for this work

Department of Plant and Environmental Sciences (Natural Sciences)

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

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.

Original language	English
Pages (from-to)	471-476
Number of pages	6
Journal	Extremophiles
Volume	12
Issue number	3
DOIs	https://doi.org/10.1007/s00792-007-0133-7
State	Published - May 2008

Bibliographical note

Funding Information:
Acknowledgments This work was supported by the grants from the Major State Basic Research Development Program of China (973 Program) (Grant No. 2004CB719604-3) and the National Natural Science Foundation of China (Grant No. 30670048).

Keywords

Alcohol dehydrogenase
Haloalkaliphilic
Molecular cloning
Natronomonas pharaonis

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10.1007/s00792-007-0133-7

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title = "Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis",

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 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.",

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author = "Yi Cao and Li Liao and Xu, {Xue Wei} and Aharon Oren and Ce Wang and Zhu, {Xu Feng} and Min Wu",

note = "Funding Information: Acknowledgments This work was supported by the grants from the Major State Basic Research Development Program of China (973 Program) (Grant No. 2004CB719604-3) and the National Natural Science Foundation of China (Grant No. 30670048).",

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doi = "10.1007/s00792-007-0133-7",

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AU - Cao, Yi

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AU - Oren, Aharon

AU - Wang, Ce

AU - Zhu, Xu Feng

AU - Wu, Min

N1 - Funding Information: Acknowledgments This work was supported by the grants from the Major State Basic Research Development Program of China (973 Program) (Grant No. 2004CB719604-3) and the National Natural Science Foundation of China (Grant No. 30670048).

PY - 2008/5

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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.

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Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis

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