【摘要】 光合细菌 C.vinosum可溶性氢酶由 52 k Da和 2 1.5k Da两个亚基组成 .该酶大亚基 N-末端氨基酸序列为 :SRTITIEPVTRXEGHAR,小亚基 N末端氨基酸序列为 :STQPKITVATXL DG.根椐大、小亚基 N-末端氨基酸序列设计两套引物 ,Primer1:5′g AT(g/ C) g T g AT(g/ C) g T(g/ A) Cg3′和 Primer2 :5′A g C AC(C/ g) CAg CC(C/ g) AA(g/ A) ATC3′,通过 PCR扩增获得 0 .6 kb的扩增片段 .克隆并对该片段序列进行分析 ,结果表明 ,该片段包含小亚基基因的全序列 .可溶性氢酶小亚基由 179个氨基酸残基组成 ,分子量为 2 1.8k Da,含有 Ni Fe-氢酶保守的序列框 :Cxx C......Gx Cxxx G......GCPP更多还原

【Abstract】 The soluble hydrogenase from C.vinosum was consist of two subunits (52 kDa, 21.5 kDa). According to the N terminal sequences of the large subunit: N’ SRTITIEPVTRXEGHAR and the small subunit: N’ STQPKITVATXLDG, Primer 1: 5’ gAT (g/C)gT gAT (g/C)gT (g/A)Cg 3’ and Primer 2: 5’ A gC AC(C/g) CAg CC(C/g) AA(g/A) ATC 3’ were synthesized and applied for PCR to amplify the fragment encoding the small subunit. The sequence analysis results showed that the soluble hydrogenase contained the reserved motif: CxxC......GxCxxxG......GCPP, which existed in all NiFe hydrogenases. C.vinosum soluble hydrogenase is a new NiFe hydrogenase which contained only one [4Fe 4S].更多还原