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首页> 外文期刊>International Journal of Molecular Sciences >Catalytic Mechanism of Short Ethoxy Chain Nonylphenol Dehydrogenase Belonging to a Polyethylene Glycol Dehydrogenase Group in the GMC Oxidoreductase Family
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Catalytic Mechanism of Short Ethoxy Chain Nonylphenol Dehydrogenase Belonging to a Polyethylene Glycol Dehydrogenase Group in the GMC Oxidoreductase Family

机译：GMC氧化还原酶家族中属于聚乙二醇脱氢酶基团的短乙氧基壬基酚脱氢酶的催化机理

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摘要

Ethoxy (EO) chain nonylphenol dehydrogenase (NPEO-DH) from Ensifer sp. AS08 and EO chain octylphenol dehydrogenase from Pseudomonas putida share common molecular characteristics with polyethylene glycol (PEG) dehydrogenases (PEG-DH) and comprise a PEG-DH subgroup in the family of glucose-methanol-choline (GMC) oxidoreductases that includes glucose/alcohol oxidase and glucose/choline dehydrogenase. Three-dimensional (3D) molecular modeling suggested that differences in the size, secondary structure and hydropathy in the active site caused differences in their substrate specificities toward EO chain alkylphenols and free PEGs. Based on 3D molecular modeling, site-directed mutagenesis was utilized to introduce mutations into potential catalytic residues of NPEO-DH. From steady state and rapid kinetic characterization of wild type and mutant NPEO-DHs, we can conclude that His465 and Asn507 are directly involved in the catalysis. Asn507 mediates the transfer of proton from a substrate to FAD and His465 transfers the same proton from the reduced flavin to an electron acceptor.
机译：来自Ensifer sp。的乙氧基（EO）链壬基酚脱氢酶（NPEO-DH）。恶臭假单胞菌的AS08和EO链辛基苯酚脱氢酶与聚乙二醇（PEG）脱氢酶（PEG-DH）具有共同的分子特征，并且在葡萄糖-甲醇-胆碱（GMC）氧化还原酶家族中包括一个PEG-DH亚组，其中包括葡萄糖/醇氧化酶和葡萄糖/胆碱脱氢酶。三维（3D）分子建模表明，活性位点的大小，二级结构和亲水性差异导致它们对EO链烷基酚和游离PEG的底物特异性不同。基于3D分子建模，利用定点诱变将突变引入NPEO-DH的潜在催化残基中。从野生型和突变型NPEO-DHs的稳态和快速动力学特征，我们可以得出结论His465和Asn507直接参与催化。 Asn507介导质子从底物到FAD的转移，而His465将相同的质子从还原的黄素转移到电子受体。

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