Abstract
S-adenosylmethionine synthetase (SAM-s) catalyzes the synthesis of S-adenosylmethionine (SAM), which is essential for methylation, transcription, proliferation, and production of secondary metabolites. Here SAM-s from Pichia ciferrii were selectively cloned using RNA CapFishing and rapid amplification of cDNA ends (RACE). The putative full-length cDNA of SAM-s encoded a 383 amino acid protein (42.6 kDa), which has highly conserved metal binding sites, a phosphate-binding site, and functionally important motifs. The corresponding enzyme was over-expressed in a heterologous host of Pichia pastoris, and then purified to a homogenous form. Enzyme kinetics, immunoblotting, circular dichroism (CD), high performance liquid chromatography (HPLC), and molecular modeling were conducted to characterize the SAM-s from P. ciferrii. Structural and functional studies of SAM-s will provide important insights for industrial applications.
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This work was supported by Grant No. R01-2003-000-10283-0 from the Basic Research Program, Korea Science and Engineering Foundation. T.D.K. is supported by a Korean Research Foundation Grant funded by the Korean Government (KRF-2009-0089832) and by a Research Grant from Ajou University.
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Yoon, S., Lee, W., Kim, M. et al. Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii . Bioprocess Biosyst Eng 35, 173–181 (2012). https://doi.org/10.1007/s00449-011-0640-x
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DOI: https://doi.org/10.1007/s00449-011-0640-x