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A Cobalt-Containing Eukaryotic Nitrile Hydratase, Salette Martinez, Xinhang Yang, Brian Bennett, Richard C. Holz
A Cobalt-Containing Eukaryotic Nitrile Hydratase, Salette Martinez, Xinhang Yang, Brian Bennett, Richard C. Holz
Physics Faculty Research and Publications
Nitrile hydratase (NHase), an industrially important enzyme that catalyzes the hydration of nitriles to their corresponding amides, has only been characterized from prokaryotic microbes. The putative NHase from the eukaryotic unicellular choanoflagellate organism Monosiga brevicollis (MbNHase) was heterologously expressed in Escherichia coli. The resulting enzyme expressed as a single polypeptide with fused α- and β-subunits linked by a seventeen-histidine region. Size-exclusion chromatography indicated that MbNHase exists primarily as an (αβ)2 homodimer in solution, analogous to the α2β2 homotetramer architecture observed for prokaryotic NHases. The NHase enzyme contained its full complement of Co(III) …