Rousset M, Dermoun Z, Hatchikian C E, Bélaich J P
Laboratoire de Chimie Bacterienne, CNRS, Marseille, France.
Gene. 1990 Sep 28;94(1):95-101. doi: 10.1016/0378-1119(90)90473-5.
The genetic locus encoding the periplasmic [NiFe]hydrogenase (Hyd) from Desulfovibrio fructosovorans was cloned and sequenced. The genes of this two-subunit enzyme have an operon organization in which the 0.94-kb gene encoding the small subunit precedes the 1.69-kb gene encoding the large subunit. A Shine-Dalgarno sequence is centered at -9 bp from the ATG of both subunits. The possible presence of another open reading frame downstream from the large-subunit-encoding gene is considered. The N-terminal sequence of the large 61-kDa subunit deduced from the nucleotide sequence is in perfect agreement with the results of the amino acid (aa) sequence determined by Edman degradation. A 50-aa leader peptide precedes the small 28-kDa subunit. The aa sequence of the enzyme shows nearly 65% homology with the [NiFe]Hyd aa sequence of Desulfovibrio gigas. Comparisons with a large range of Hyds from various bacterial species indicate the presence of highly conserved Cys residues, the implications of which are discussed from the point of view of nickel atom and cluster accommodation.
编码来自果糖脱硫弧菌周质[NiFe]氢化酶（Hyd）的基因座被克隆并测序。这种双亚基酶的基因具有操纵子结构，其中编码小亚基的0.94 kb基因位于编码大亚基的1.69 kb基因之前。一个Shine-Dalgarno序列位于两个亚基ATG上游9 bp处。考虑了在编码大亚基的基因下游可能存在的另一个开放阅读框。从核苷酸序列推导的61 kDa大亚基的N端序列与通过埃德曼降解确定的氨基酸（aa）序列结果完全一致。一个50个氨基酸的前导肽位于28 kDa小亚基之前。该酶的氨基酸序列与巨大脱硫弧菌的[NiFe]Hyd氨基酸序列显示出近65%的同源性。与来自各种细菌物种的大量氢化酶的比较表明存在高度保守的半胱氨酸残基，并从镍原子和簇容纳的角度讨论了其意义。
