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首页> 外文期刊>Biochemistry >Structural and Kinetic Properties of the Aldehyde Dehydrogenase NahF, a Broad Substrate Specificity Enzyme for Aldehyde Oxidation
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Structural and Kinetic Properties of the Aldehyde Dehydrogenase NahF, a Broad Substrate Specificity Enzyme for Aldehyde Oxidation

机译：醛脱氢酶NahF，醛氧化的广泛底物特异性酶的结构和动力学性质。

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摘要

The salicylaldehyde dehydrogenase (NahF) catalyzes the oxidation of salicylaldehyde to salicylate using NAD(+) as a cofactor, the last reaction of the upper degradation pathway of naphthalene in Pseudomonas putida G7. The naphthalene is an abundant and toxic compound in oil and has been used as a Model for bioremediation studies. The steady-state kinetic parameters for oxidation of aliphatic or aromatic aldehydes catalyzed by 6xHis-NahF are presented. The 6xHis-NahF catalyzes the oxidation of aromatic aldehydes with large k(cat)/K-m values close to 10(6) M-1 s(-1). The active site of NahF is highly hydrophobic, and the enzyme shows higher specificity for less polar substrates than for polar substrates, e.g., acetaldehyde. The enzyme shows alpha/beta folding with three well-defined domains: the oligomerization domain, which is responsible for the interlacement between the two monomers; the Rossmann-like fold domain, essential for nucleotide binding; and the catalytic domain. A salicylaldehyde molecule was Observed in a deep pocket in the crystal structure of NahF where the catalytic C284 and E250 are present. Moreover, the residues G150, R157, W96, F99, F274, F279, and Y446 were thought to be important for catalysis and specificity for aromatic aldehydes. Understanding the molecular features responsible for NahF activity allows for comparisons with other aldehyde dehydrogenases and, together with structural information, provides the information needed for future mutational studies aimed to, enhance its stability and specificity and further its use in biotechnological processes.
机译：水杨醛脱氢酶（NahF）使用NAD（+）作为辅因子催化水杨醛氧化为水杨酸酯，这是恶臭假单胞菌G7中萘上级降解途径的最后反应。萘是石油中一种丰富而有毒的化合物，已被用作生物修复研究的模型。提出了由6xHis-NahF催化的脂族或芳族醛氧化的稳态动力学参数。 6xHis-NahF催化k（cat）/ K-m值接近10（6）M-1 s（-1）的芳香醛的氧化。 NahF的活性位点是高度疏水的，并且该酶对极性较小的底物显示出比对极性底物例如乙醛更高的特异性。该酶显示具有三个明确定义的域的α/β折叠：寡聚域，负责两个单体之间的交错; Rossmann样折叠域，对核苷酸结合必不可少;和催化领域。在NahF晶体结构的一个深口袋中观察到了水杨醛分子，其中存在催化C284和E250。此外，残基G150，R157，W96，F99，F274，F279和Y446被认为对芳香醛的催化和特异性很重要。了解负责NahF活性的分子特征可以与其他醛脱氢酶进行比较，并与结构信息一起，为未来的突变研究提供了所需的信息，旨在增强其稳定性和特异性，并进一步用于生物技术过程。

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