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首页> 外文期刊>Biochemistry >Heterologous Production of Halorhodospira halophila Holo-Photoactive Yellow Protein through Tandem Expression of the Postulated Biosynthetic Genes
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Heterologous Production of Halorhodospira halophila Holo-Photoactive Yellow Protein through Tandem Expression of the Postulated Biosynthetic Genes

机译:Halorhodospira Halophila Holo-Photo活性黄蛋白的异源生产通过串联生物合成基因的表达

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摘要

The photoactive yellow protein (PYP) is a bacterial photoreactor which is the structural prototype for the PAS domain superfamily of regulators and receptors. PYP is known to have a unique p-hydroxycinnamic acid chromophore, covalently attached to a cysteine. To date, it has not been shown how holo-PYP is formed in vivo. Two genes, nearby pyp, were postulated to encode the biosnthetic enzymes, but only one was previously isolated and shown to have the requisite activity. By using a dual plasmid system, one expressing the PYP from Halorhodospira halophila and the other expressing a two-gene operon, consisting of tyrosine ammonia lyase and p-hydroxycinnamic acid ligase, we are able to present evidence that a functionally active holo-PYP can be synthesized in Escherichia coli. Plasmids containing only one of the two enzymes failed to produce holoprotein. Thus, the two genes have been shown to be both necessary and sufficient for production of holoprotein, although the activating group remains unknown. This expression system not only holds great potential for mutagenesis studies but also opens new possibilities in the search for (a) signaling partner(s) of the PYP.
机译:光活性黄蛋白(PYP)是细菌光反应器,其是调节剂和受体的PAS结构域的结构原型。已知本谱具有独特的羟基氨基酸发色团,共价连接到半胱氨酸。迄今为止,尚未显示Holo-Pyp如何在体内形成。假设两个基因,附近的PYP,以编码生物染色酶,但先前只分离出一个并显示有必要的活性。通过使用双重质粒系统,从Halorhodospira Halophila表达PYP,另一个表达双基因操纵子,由酪氨酸氨裂解酶和对羟基氨基酸连接酶组成,我们能够呈现功能性活跃的Holo-pyp在大肠杆菌中合成。含有两种酶中的一种的质粒未能产生全蛋白酶。因此,两种基因已被证明是必要的并且足以用于生产全蛋白质,尽管活化组仍然未知。此表达系统不仅具有诱变研究的巨大潜力,而且还可以在搜索PYP的(a)信令伙伴的搜索中开启新的可能性。

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  • 来源
    《Biochemistry》 |2003年第4期|共6页
  • 作者单位

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

    Department of Protein Biochemistry and Protein Engineering University of Gent Ledegankstraat 35 9000 Gent Belgium and Department of Biochemistrty and Molecular Biophysics University of Arzona Tucson Arizona 85721;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
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