Combining a Nitrogenase Scaffold and a Synthetic Compound into an Artificial Enzyme.
Combining a Nitrogenase Scaffold and a Synthetic Compound into an Artificial Enzyme.
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DOI：
10.1002/anie.201507646
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发表时间：
2015-11-16
期刊：
影响因子：
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通讯作者：
Ribbe MW
中科院分区：
文献类型：
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作者：
Tanifuji K;Lee CC;Ohki Y;Tatsumi K;Hu Y;Ribbe MW
固氮酶在其辅因子中心（[（Cit）MoFe 7S 9 C]n−;命名为M簇）催化底物还原。在这里，我们报告了将合成模型复合物（[Fe 6S 9（SEt）2]4-;指定为Fe 6 RHH）插入到固氮酶的催化组分（指定为NifDK）中后，形成了一种人工的、模拟固氮酶的酶。将两个Fe 6 RHH簇插入NifDK中，使得所得蛋白质（命名为NifDKFe）处于与插入天然M簇时的构象相似的构象。NifDKFe可以与固氮酶的还原酶组分一起在ATP依赖的反应中还原C2 H2。在Eu（II）DTPA存在下，它也可以作为一种酶，在C2 H2还原中表现出很强的活性，同时证明了以ATP非依赖性方式将CN−还原为C1-C3烃的能力。这项工作的成功结果为通过这种方法继续寻找新型酶活性提供了概念证明和基本原则。
Nitrogenase catalyzes substrate reduction at its cofactor center ([(Cit)MoFe7S9C]n−; designated M-cluster). Here, we report the formation of an artificial, nitrogenase-mimicking enzyme upon insertion of a synthetic model complex ([Fe6S9(SEt)2]4−; designated Fe6RHH) into the catalytic component of nitrogenase (designated NifDK). Two Fe6RHH clusters were inserted into NifDK, rendering the resultant protein (designated NifDKFe) in a similar conformation to that upon insertion of native M-clusters. NifDKFe could work together with the reductase component of nitrogenase to reduce C2H2 in an ATP-dependent reaction. It could also act as an enzyme on its own in the presence of Eu(II) DTPA, displaying a strong activity in C2H2 reduction while demonstrating an ability to reduce CN− to C1-C3 hydrocarbons in an ATP-independent manner. The successful outcome of this work provides the proof of concept and underlying principles for continued search of novel enzymatic activities via this approach.