Journal ArticleDOI
Purification of Penicillin G Amidase using Quaternary Ammonium Salts and effect on the activity of the immobilised enzymes
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TLDR
It was concluded that the quaternary ammonium salts used selectively precipitated the non-enzymatic protein leaving in solution practically all the enzyme resulting in a high yield of purification.Abstract:
This paper describes the purification of Penicillin G Amidase (EC 3.5.1.11) using quaternary ammonium salts with the aim of increasing the activity of immobilised enzymes prepared from the purified solutions. Two different quaternary ammonium salts were tested with different solutions of the enzyme. It was concluded that the quaternary ammonium salts used selectively precipitated the non-enzymatic protein leaving in solution practically all the enzyme resulting in a high yield of purification. Optimal conditions for purification using the two types of quaternary ammonium salts were determined. Immobilisation studies were performed from various purified enzyme solutions, using different amounts of a quaternary ammonium salt. The immobilised enzymes so obtained showed a much higher activity than the immobilised enzyme obtained from non-purified enzyme solutions.read more
Citations
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Journal ArticleDOI
Optimisation of penicillin acylase extraction by AOT/isooctane reversed micellar systems
TL;DR: In this article, Escherichia coli penicillin acylase was successfully extracted from a crude extract obtained by osmotic shock into the reversed micelle phase of the anionic surfactant AOT in isooctane and back-extracted, almost pure, to a final aqueous phase by simple salt concentration change.
Journal ArticleDOI
Integrated L-phenylalanine separation in an E. coli fed-batch process: from laboratory to pilot scale.
TL;DR: Pilot-scale reactive-extraction technology for fully integrated L-phenylalanine separation in Escherichia coli fed-batch fermentations was investigated in order to prevent an inhibition of microbial L-Phe production by-product accumulation.
Journal ArticleDOI
An integrated downstream processing strategy for the recovery and partial purification of penicillin acylase from crude media
TL;DR: An integrated process strategy for the recovery of penicillin acylase was developed, based on precipitation of non-enzymatic proteins directly from Escherichia coli homogenates or crude extracts using Rolquat and adsorption of the enzyme on Amp-Seph under pseudo-affinity conditions.
References
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Journal Article
Protein Measurement with the Folin Phenol Reagent
TL;DR: Procedures are described for measuring protein in solution or after precipitation with acids or other agents, and for the determination of as little as 0.2 gamma of protein.
Journal ArticleDOI
The isolation and kinetics of penicillin amidase from Escherichia coli
TL;DR: The enzyme was shown to be inhibited by excess substrate, benzylpenicillin, and by both of the products of hydrolysis, which was found to be competitive and by 6-aminopenicillanic acid to be non-competitive.
Book ChapterDOI
Penicillin acylase (bacterial).
T A Savidge,M Cole +1 more
TL;DR: The published work on the preparation and properties of the extracellular enzyme produced by the gram-positive organism Bacillus megaterium is presented and the following two methods are recommended: Crystalline enzyme has been obtained by the second method.