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The lack of activity will not be because of a diminished ability to bind divalent cations Because the steel articles from the purified mutants was just like the wild sort enzyme. The catalytic properties of such mutants are absolutely in line with the proposed part of the residue in the chemical mechanism as the main basic base for activation from the hydrolytic water molecule and the next protonation of the leaving group amine.
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A tetrahedral intermediate is shaped and afterwards collapses with cleavage on the C-N bond right after proton transfer for the leaving team amine by Asp-273. The lack of a solvent isotope influence by D2O as well as the absence of any improvements on the kinetic constants with increases in solvent viscosity suggest that Web product formation is not really limited to any important extent by proton transfer actions or the release of items. N
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