Cloning and sequencing of a [NiFe] hydrogenase operon from Desulfovibrio vulgaris Miyazaki F.
Cloning and sequencing of a [NiFe] hydrogenase operon from Desulfovibrio vulgaris Miyazaki F.
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来自普通脱硫弧菌 Miyazaki F 的 [NiFe] 氢化酶操纵子的克隆和测序。
DOI：
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发表时间：
1990
期刊：
影响因子：
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通讯作者：
G. Voordouw
中科院分区：
文献类型：
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作者：
H. M. Deckers;F. R. Wilson;G. Voordouw
利用普通脱硫弧菌（Desulfovibrio vulgaris Miyazaki F） [NiFe]氢化酶编码基因探针，克隆了宫崎脱硫弧菌（Desulfovibrio vulgaris Hildenborough）氢化酶操纵子。克隆DNA的核酸序列表明，这种氢化酶是一种双亚基酶：小亚基（267个残基，分子质量= 28763 Da）的基因先于大亚基（566个残基，分子质量= 62495 Da）的基因，与其他[NiFeSe]和[NiFeSe]氢化酶操纵子一样。Miyazaki氢化酶的大小亚基氨基酸序列与Desulfovibrio gigas的[NiFe]氢化酶具有80%的同源性。14个半胱氨酸残基，10个在小亚基上，4个在大亚基上，被认为在[NiFe]氢化酶中协调铁硫簇和活性位点镍，在宫崎氢化酶中被发现是保守的。该基因序列的亚基分子质量和氨基酸组成与Yagi及其同事分离的质周膜结合氢化酶的数据非常相似，这表明该氢化酶属于[NiFe]氢化酶的一般类别，尽管其镍含量低且光谱性质明显异常。
A hydrogenase operon was cloned from chromosomal DNA isolated from Desulfovibrio vulgaris Miyazaki F with the use of probes derived from the genes encoding [NiFe] hydrogenase from Desulfovibrio vulgaris Hildenborough. The nucleic acid sequence of the cloned DNA indicates this hydrogenase to be a two-subunit enzyme: the gene for the small subunit (267 residues; molecular mass = 28763 Da) precedes that for the large subunit (566 residues; molecular mass = 62495 Da), as in other [NiFe] and [NiFeSe] hydrogenase operons. The amino acid sequences of the small and large subunits of the Miyazaki hydrogenase share 80% homology with those of the [NiFe] hydrogenase from Desulfovibrio gigas. Fourteen cysteine residues, ten in the small and four in the large subunit, which are thought to co-ordinate the iron-sulphur clusters and the active-site nickel in [NiFe] hydrogenases, are found to be conserved in the Miyazaki hydrogenase. The subunit molecular masses and amino acid composition derived from the gene sequence are very similar to the data reported for the periplasmic, membrane-bound hydrogenase isolated by Yagi and coworkers, suggesting that this hydrogenase belongs to the general class of [NiFe] hydrogenases, despite its low nickel content and apparently anomalous spectral properties.