Purification and Properties of Aryol Acylamidase from Pseudomonas fluorescens ATCC 39004

Open Access
Publisher Website
Google Scholar
Abstract
Aryl acytlamidase 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 52500 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 806 and at a temperature of 45°C. The enzyme shows Michaelis-Menten kinetics; Km vlaues for nitroacetanilide (69μM) and hydroxyacetanilide (601μM) were low, indicating that the enzyme has a very high affinity for both substrates.