巨大脱硫弧菌[NiFe]氢化酶在果糖脱硫弧菌MR400中的异源表达。
Heterologous expression of the Desulfovibrio gigas [NiFe] hydrogenase in Desulfovibrio fructosovorans MR400.
作者信息
Rousset M, Magro V, Forget N, Guigliarelli B, Belaich J P, Hatchikian E C
机构信息
Unité de Bioénergétique et Ingénierie des Protéines, IBSM, CNRS, 13402 Marseille Cedex 20, France.
出版信息
J Bacteriol. 1998 Sep;180(18):4982-6. doi: 10.1128/JB.180.18.4982-4986.1998.
Abstract
The ability of Desulfovibrio fructosovorans MR400 DeltahynABC to express the heterologous cloned [NiFe] hydrogenase of Desulfovibrio gigas was investigated. The [NiFe] hydrogenase operon from D. gigas, hynABCD, was cloned, sequenced, and introduced into D. fructosovorans MR400. A portion of the recombinant heterologous [NiFe] hydrogenase was totally matured, exhibiting catalytic and spectroscopic properties identical to those of the native D. gigas protein. A chimeric operon containing hynAB from D. gigas and hynC from D. fructosovorans placed under the control of the D. fructosovorans hynAp promoter was constructed and expressed in D. fructosovorans MR400. Under these conditions, the same level of activity was obtained as with the D. gigas hydrogenase operon.
摘要
研究了果糖脱硫弧菌MR400 DeltahynABC表达巨大脱硫弧菌异源克隆的[NiFe]氢化酶的能力。克隆、测序了来自巨大脱硫弧菌的[NiFe]氢化酶操纵子hynABCD，并将其导入果糖脱硫弧菌MR400。重组异源[NiFe]氢化酶的一部分完全成熟，表现出与天然巨大脱硫弧菌蛋白相同的催化和光谱性质。构建了一个嵌合操纵子，其包含来自巨大脱硫弧菌的hynAB和来自果糖脱硫弧菌的hynC，并置于果糖脱硫弧菌hynAp启动子的控制下，在果糖脱硫弧菌MR400中表达。在这些条件下，获得了与巨大脱硫弧菌氢化酶操纵子相同水平的活性。
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