Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of an O-Methyltransferase from Streptomyces avermitilis MA-4680

  • Yoon, Young-Dae (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Park, Young-Hee (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Yi, Yong-Sub (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Lee, Young-Shim (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Jo, Geun-Hyeong (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Park, Jun-Cheol (National Institute of Animal Science, Rural Development Administration) ;
  • Ahn, Joong-Hoon (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University) ;
  • Lim, Yoong-Ho (Division of Bioscience and Biotechnology, BMIC, RCD, Konkuk University)
  • Received : 2010.05.10
  • Accepted : 2010.06.01
  • Published : 2010.09.28
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Abstract

A search of the Streptomyces avermitilis genome reveals that its closest homologs are several O-methyltransferases. Among them, one gene (viz., saomt5) was cloned into the pET-15b expression vector by polymerase chain reaction using sequence-specific oligonucleotide primers. Biochemical characterization with the recombinant protein showed that SaOMT5 was S-adenosyl-L-methionine-dependent Omethyltransferase. Several compounds were tested as substrates of SaOMT5. As a result, SaOMT5 catalyzed O-methylation of flavonoids such as 6,7-dihydroxyflavone, 2',3'-dihydroxyflavone, 3',4'-dihydroxyflavone, quercetin, and 7,8-dihydroxyflavone, and phenolic compounds such as caffeic acid and caffeoyl Co-A. These reaction products were analyzed by TLC, HPLC, LC/MS, and NMR spectroscopy. In addition, SaOMT5 could convert phenolic compounds containing ortho-dihydroxy groups into O-methylated compounds, and 6,7-dihydroxyflavone was known to be the best substrate. SaOMT5 converted 6,7-dihydroxyflavone into 6-hydroxy-7-methoxyflavone and 7-hydroxy-6-methoxyflavone, and caffeic acid into ferulic acid and isoferulic acid, respectively. Moreover, SaOMT5 turned out to be a $Mg^{2+}$-dependent OMT, and the effect of $Mg^{2+}$ ion on its activity was five times greater than those of $Ca^{2+}$, $Fe^{2+}$, and $Cu^{2+}$ ions, EDTA, and metal-free medium.

Keywords

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