Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Purification, Characterization, and Cloning of Trimethylamine Dehydrogenase from Methylophaga sp. Strain SK1

  • Kim, Hee-Gon (Department of Biomaterials Engineering, Chosun University) ;
  • Kim, Yan (Department of Biomaterials Engineering, Chosun University) ;
  • Lim, Heon-Man (Department of Biology, Chungnam National University) ;
  • Shin, Hyun-Jae (Department of Chemical Engineering, Chosun University) ;
  • Kim, Si-Wouk (Department of Biomaterials Engineering, Chosun University)
  • Published : 2006.08.30
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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 from Methylophaga 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 $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga 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 of Methylophilus methylotrophus $W_3A_1$.

Keywords

References

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