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Examination Of The Catalytic Role Of The Axial Cystine Ligand In The Co-Type Nitrile Hydratase From Pseudonocardia Thermophila Jcm 3095, Irene Anyango Ogutu, Martin St. Maurice, Brian Bennett, Richard C. Holz
Examination Of The Catalytic Role Of The Axial Cystine Ligand In The Co-Type Nitrile Hydratase From Pseudonocardia Thermophila Jcm 3095, Irene Anyango Ogutu, Martin St. Maurice, Brian Bennett, Richard C. Holz
Physics Faculty Research and Publications
The strictly conserved αSer162 residue in the Co-type nitrile hydratase from Pseudonocardia thermophila JCM 3095 (PtNHase), which forms a hydrogen bond to the axial αCys108-S atom, was mutated into an Ala residue. The αSer162Ala yielded two different protein species: one was the apoform (αSerA) that exhibited no observable activity, and the second (αSerB) contained its full complement of cobalt ions and was active with a kcat value of 63 ± 3 s−1 towards acrylonitrile at pH 7.5. The X-ray crystal structure of was determined at 1.85 Å resolution and contained no detectable …