title: Isolation and primary characterization of an amidase from Rhodococcus rhodochrous.
abstract: Amidase (EC 3.5.1.4) was purified to homogeneity from Rhodococcus rhodochrous M8 using isopropanol fractionation and exchange chromatography on Mono Q. The isolated amidase consists of four identical subunits with molecular weight 42+/-2 kD. The activity of the enzyme is maximal at 55-60 degrees C and within the pH range 5-8. The amidase from R. rhodochrous M8 is highly sensitive to such sulfhydryl reagents as Hg2+ and Cu2+. Chelators (EDTA and o-phenanthroline) and serine proteinase inhibitors (PMSF and DIFP) did not inhibit the activity of the enzyme. The enzyme exhibits hydrolytic and acyl transferase activity and does not possess urease activity. Aliphatic amides (acetamide and propionamide) were the best substrates for the amidase from R. rhodochrous M8, whereas bulky aromatic amides were poor substrates of this enzyme. The properties of the isolated enzyme are similar to those found in the corresponding amidase from Arthrobacter sp. J-1 and an amidase with wide substrate specificity from Brevibacterium sp. R312.
authors.affiliation: Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 113545, Russia. lab11@vnigen.msk.su.
authors.forename: E K
authors.initials: EK
authors.lastname: Kotlova
authors.forename: G G
authors.initials: GG
authors.lastname: Chestukhina
authors.forename: O B
authors.initials: OB
authors.lastname: Astaurova
authors.forename: T E
authors.initials: TE
authors.lastname: Leonova
authors.forename: A S
authors.initials: AS
authors.lastname: Yanenko
authors.forename: V G
authors.initials: VG
authors.lastname: Debabov
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008670:Metals / Q000494:pharmacology; D012240:Rhodococcus / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000596:Amino Acids; D004791:Enzyme Inhibitors; D008670:Metals; D000581:Amidohydrolases; C019665:amidase