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Purification, characterization, and cloning of trimethylamine dehydrogenase fromMethylophaga sp. strain SK1

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Abstract

Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified fromMethylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and 55°C, respectively. TheV max andK m values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp fromMethylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that ofMethylophilus methylotrophus W3A1.

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Authors and Affiliations

  1. Department of Biomaterials Engineering, Chosun University, 501-759, Gwangju, Korea

    Hee Gon Kim, Yan Kim & Si Wouk Kim

  2. Department of Biology, Chungnam National University, 305-764, Daejon, Korea

    Heon Man Lim

  3. Department of Chemical Engineering, Chosun University, 501-759, Gwangju, Korea

    Hyun-Jae Shin

  4. Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University, 501-759, Gwangju, Korea

    Si Wouk Kim

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  1. Hee Gon Kim
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Correspondence to Si Wouk Kim.

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Kim, H.G., Kim, Y., Lim, H.M. et al. Purification, characterization, and cloning of trimethylamine dehydrogenase fromMethylophaga sp. strain SK1. Biotechnol. Bioprocess Eng. 11, 337–343 (2006). https://doi.org/10.1007/BF03026250

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  • DOI: https://doi.org/10.1007/BF03026250

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