title: Aliphatic amidase from Rhodococcus rhodochrous M8 is related to the nitrilase/cyanide hydratase family.
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.
authors.affiliation: Faculty of Bioengineering and Bioinformatics, Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119992, Russia.
authors.forename: S I
authors.initials: SI
authors.lastname: Pertsovich
authors.forename: D T
authors.initials: DT
authors.lastname: Guranda
authors.forename: D A
authors.initials: DA
authors.lastname: Podchernyaev
authors.forename: A S
authors.initials: AS
authors.lastname: Yanenko
authors.forename: V K
authors.initials: VK
authors.lastname: Svedas
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000619:Aminohydrolases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006836:Hydro-Lyases / Q000378:metabolism*; D008969:Molecular Sequence Data; D012240:Rhodococcus / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
chemical_list: D000581:Amidohydrolases; C019665:amidase; D000619:Aminohydrolases; C021867:nitrilase; D006836:Hydro-Lyases; C076848:cyanide hydratase