Penicillin amidase

About: Penicillin amidase is a research topic. Over the lifetime, 576 publications have been published within this topic receiving 15563 citations. The topic is also known as: penicillin amidohydrolase & ampicillin acylase.

Separation and purification of penicillin acylase from Escherichia coli using AOT reverse micelles

Abstract: The extraction of penicillin acylase by reverse micellar solutions of a surfactant was studied. A 50 mM solution of dioctyl sodium sulphosuccinate in isooctane extracted 46% of the enzyme activity in a crude periplasmic extract of induced cells of E. coli ATCC 9637. The increase in the specific activity of the final enzyme preparation, after stripping of the organic phase at pH 7.5, in the presence of 1 M KCl, was 8 - fold.

Modified fluorimetric assay for estimating ampicilloate concentrations and its use for detecting β-lactamase and penicillin acylase activity in bacteria

Abstract: Sodium ampicilloate concentrations were estimated fluorimetrically by heating solutions with ascorbic acid, EDTA and a modified Lowry A reagent which was prepared by including copper sulfate and potassium sodium tartrate in 0.5 mol dm - 3 acetate buffer at pH 4. A concentration range of 0.5–50 µmol dm - 3 was used for the estimations. The reaction was used to estimate β-lactamase activity on ampicillin but the substrate also showed some fluorescence and a calculation was required to determine the amount of ampicilloate formed when both substances were present in the one reaction mixture. The β-lactamase was inhibited by treatment with trichloroacetic acid so the procedure could be used to assay the enzyme activity after a fixed time. 6-Aminopenicillanic acid did not fluoresce on treatment with the modified reagent and organisms which contained penicillin acylase lowered the amount of ampicillin which could be converted to ampicilloate. When penicillin acylase and β-lactamase co-existed in the one organism, the respective activities were determined by use of the copper–ascorbate–EDTA fluorescence assay for ampicilloate coupled with a fluorescamine assay for 6-aminopenicillanic acid determinations. On prolonged incubation, some organisms containing penicillin acylases lowered the amount of ampicilloate which formed a fluorescent product. This effect was attributed to deacylation of ampicilloate by the penicillin acylases.

A homology model of penicillin acylase from Alcaligenes faecalis and in silico evaluation of its selectivity.

Abstract: A three-dimensional model of the relatively unknown penicillin acylase from Alcaligenes faecalis (PA-AF) was built up by means of homology modeling based on three different crystal structures of penicillin acylase from various sources. An in silico selectivity study was performed to compare this homology model to the structure of the Escherichia coli enzyme (PA-EC) in order to find any selectivity differences between the two enzymes. The program GRID was applied in combination with the principal component analysis technique to identify the regions of the active sites where the PAs potentially engage different interactions with ligands. These differences were further analyzed and confirmed by molecular docking simulations. The PA-AF homology model provided the structural basis for the explanation of the different enantioselectivities of the enzymes previously demonstrated experimentally and reported in the literature. Different substrate selectivities were also predicted for PA-AF compared to PA-EC. Since no crystallographic data are available for PA-AF to date, the three-dimensional homology model represents a useful and efficient tool for fully exploiting this attractive and efficient biocatalyst, particularly in enantioselective acylations of amines.