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首页> 外文期刊>Bioprocess and Biosystems Engineering >Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii
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Structural and functional characterization of S-adenosylmethionine (SAM) synthetase from Pichia ciferrii

机译：毕赤酵母中S-腺苷甲硫氨酸（SAM）合成酶的结构和功能表征

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摘要

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.
机译：S-腺苷甲硫氨酸合成酶（SAM-s）催化S-腺苷甲硫氨酸（SAM）的合成，这对于甲基化，转录，增殖和次级代谢产物的产生至关重要。在这里，使用RNA CapFishing选择性克隆了来自毕赤酵母的SAM-s，并快速扩增了cDNA末端（RACE）。推测的SAM-s全长cDNA编码383个氨基酸的蛋白质（42.6 kDa），该蛋白质具有高度保守的金属结合位点，磷酸结合位点和功能重要的基序。相应的酶在巴斯德毕赤酵母的异源宿主中过表达，然后纯化为同质形式。进行了酶动力学，免疫印迹，圆二色性（CD），高效液相色谱（HPLC）和分子建模来表征来自拟南芥（P. ciferrii）的SAM-s。 SAM-s的结构和功能研究将为工业应用提供重要的见识。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2012年第2期|p.173-181|共9页
  • 作者

    Sangyoung Yoon; Wonkyu Lee; Minsoo Kim; T. Doohun Kim; Yeonwoo Ryu;

  • 作者单位

    Department of Molecular Science and Technology,Graduate School of Interdisciplinary Program,Ajou University, Suwon 443-749, Korea;

    Department of Molecular Science and Technology,Graduate School of Interdisciplinary Program,Ajou University, Suwon 443-749, Korea;

    Jeonbuk Branch Institute Bioindustry Research Center,Korea Research Institute of Bioscience and Biotechnology,Jeonbuk 580-185, Korea;

    Department of Molecular Science and Technology,Graduate School of Interdisciplinary Program,Ajou University, Suwon 443-749, Korea;

    Department of Molecular Science and Technology,Graduate School of Interdisciplinary Program,Ajou University, Suwon 443-749, Korea;

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  • 关键词

    s-adenosylmethionine; SAM synthetase; pichia ciferrii; RNA capfishing; rapid amplification of cdna ends;

    机译：s-腺苷甲硫氨酸;SAM合成酶;毕赤酵母RNA捕获;cdna末端快速扩增;

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