Purification and properties of aryl acylamidase from Pseudomonas fluorescens ATCC 39004.
Aryl acylamidase has been purified from a strain of Pseudomonas fluorescens ATCC 39004, selected from soil on the basis of its ability to utilise acylanilide compounds as a sole source of carbon. The enzyme was purified to homogeneity by a combination of ion-exchange, hydrophobic and gel-permeation chromatography. A relative molecular mass of about 52 500 was estimated by gel filtration. The native enzyme was shown to be a monomeric protein by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. The enzyme was maximally active at a pH of 8.6 and at a temperature of 45 degrees C. The enzyme shows Michaelis-Menten kinetics; Km values for nitroacetanilide (69 microM) and hydroxyacetanilide (6.1 microM) were low, indicating that the enzyme has a very high affinity for both substrates.
D000581:Amidohydrolases / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002621:Chemistry; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D011551:Pseudomonas fluorescens / Q000201:enzymology*; D013696:Temperature
D008670:Metals; D000581:Amidohydrolases; C019732:aryl acylamidase