Abstract
A comparative study of amino acid sequence and physicochemical properties indicates the affiliation of an amidase from Rhodococcus rhodochrous M8 (EC 3.5.1.4) to the nitrilase/cyanide hydratase family. Cluster analysis and multiple alignments show that Cys166 is an active site nucleophile. The enzyme has been shown to be a typical aliphatic amidase, being the most active toward short-chain linear amides. Small polar molecules such as hydroxylamine and O-methyl hydroxylamine can serve as effective external nucleophiles in acyl transfer reactions. The kinetics of the industrially important amidase-catalyzed acrylamide hydrolysis has been studied over a wide range of substrate concentrations; inhibition during enzymatic hydrolysis by the substrate and product (acrylic acid) has been observed; an adequate kinetic scheme has been evaluated and the corresponding kinetic parameters have been determined.
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Translated from Biokhimiya, Vol. 70, No. 11, 2005, pp. 1556–1565.
Original Russian Text Copyright © 2005 by Pertsovich, Guranda, Podchernyaev, Yanenko, Svedas.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM05-060, June 12, 2005.
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Pertsovich, S.I., Guranda, D.T., Podchernyaev, D.A. et al. Aliphatic Amidase from Rhodococcus rhodochrous M8 Is Related to the Nitrilase/Cyanide Hydratase Family. Biochemistry (Moscow) 70, 1280–1287 (2005). https://doi.org/10.1007/s10541-005-0260-7
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DOI: https://doi.org/10.1007/s10541-005-0260-7