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Heterologous Expression and Characterization of an Alcohol Dehydrogenase from the Archeon Thermoplasma acidophilum

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Abstract

Analysis of the Thermoplasma acidophilum DSM 1728 genome identified two putative alcohol dehydrogenase (ADH) open reading frames showing 50.4% identity against each other. The corresponding genes Ta0841 and Ta1316 encode proteins of 336 and 328 amino acids with molecular masses of 36.48 and 36.01 kDa, respectively. The genes were expressed in Escherichia coli and the recombinant enzymes were functionally assessed for activity. Throughout the study only Ta1316 ADH resulted active in the oxidative reaction in the pH range 2–8 (optimal pH 5.0) and temperatures from 25 to 90°C (optimal 75°C). This ADH catalyzes the oxidation of several alcohols such as ethanol, methanol, 2-propanol, butanol, and pentanol during the reduction of the cofactor NAD+. The highest activity was found in the presence of ethanol producing optically pure acetaldehyde. The specific enzyme activity of the purified Ta1316 ADH with ethanol as a substrate in the optimal conditions was 628.7 U/mg.

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Abbreviations

ADH:

Alcohol dehydrogenase

Orf :

Open reading frame

NAD:

Nicotin adenine dinucleotide

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Acknowledgment

Erika N. Marino-Marmolejo was supported by the Graduate Scholarship No. 182072 from the National Council for Science and Technology (CONACYT), Mexico.

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Authors and Affiliations

  1. Division of Molecular Biology, Institute for Scientific and Technological Research of San Luis Potosí (IPICYT), Camino a la Presa San Jose 2055, Lomas 4a Seccion, CP 78216, San Luis Potosi, Mexico

    Erika Nahomy Marino-Marmolejo, Antonio De León-Rodríguez, Ana Paulina Barba de la Rosa & Leticia Santos

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  1. Erika Nahomy Marino-Marmolejo
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  2. Antonio De León-Rodríguez
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Correspondence to Leticia Santos.

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Marino-Marmolejo, E.N., De León-Rodríguez, A., de la Rosa, A.P.B. et al. Heterologous Expression and Characterization of an Alcohol Dehydrogenase from the Archeon Thermoplasma acidophilum . Mol Biotechnol 42, 61–67 (2009). https://doi.org/10.1007/s12033-008-9130-z

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