Studies on improved techniques for immobilizing and stabilizing penicillin amidase associated with E. coli cells
TL;DR: The whole cell immobilization technique has been optimized for different process parameters and the granular catalyst has good mechanical strength, low protein leachability, and high retention of penicillin amidase activity.
About: This article is published in Enzyme and Microbial Technology.The article was published on 1991-08-01. It has received 24 citations till now. The article focuses on the topics: Penicillin amidase activity & Penicillin amidase.
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TL;DR: Experimental findings show that the biosynthesis of penicillin amidase in E. coli show a little variation among batch reactor modes and significant variation on the continuous mode of operation, and analysis shows that the different reactor modes also influence periplasmic localization of the enzyme in the cell.
Abstract: Different operational mode of bioreactors influence the biosynthesis of the enzyme and related products as well as the growth of industrial microorganisms. This communication deals with the effect of mode of operation of various bioreactors with different geometric configurations, viz., batch (includes commercially available batch stirred tank, and custom-designed cylindrical and tapered reactors), batch-fed, continuous flow stirred tank reactors on the biosynthesis of penicillin amidase in Escherichia coli. Experimental findings show that the biosynthesis of penicillin amidase in E. coli show a little variation among batch reactor modes and significant variation on the continuous mode of operation. Further analysis show that the different reactor modes also influence periplasmic localization of the enzyme in the cell.
2 citations
Cites methods from "Studies on improved techniques for ..."
...Estimation of cell mass The cell concentration in the suspension was determined spectrophotometrically by measuring the absorbance at 520 nm (spectrophotometer DIGISPEC 110 D, SICO, India) [ 5 ]....
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Dissertation•
30 Aug 2017
1 citations
01 Oct 2012
TL;DR: The approach is promising to develop a strategy for debittering citrus juices by biotransforming limonin at a faster rate by biOTransforming the bitter principle - Limonin.
Abstract: This study was aimed for optimizing the immobilization parameters for Pseudomonas putida G7 in Ca-alginate beads, in order to establish a debittering strategy for citrus juices, by biotransforming the bitter principle - Limonin. Response Surface Methodology (RSM) with Central Composite Design (CCD) was employed to model the significant parameters for an enhanced response. An enhanced limonin bioconversion and immobilized bead stability was obtained with alginate concentration (2%), cell load (47.2g/l), and a bead diameter (2.1mm); which had significant effects (p <0.001) on limonin biotransformation. The R2 values of 0.9 showed good agreement between experimental and predicted response. Validation experiments under optimized parameters showed good association between experimental (limonin biotransformation and stability response of 65.8% and 0.97 OD respectively) and predicted responses (limonin biotransformation and stability of 65.1% and 0.094 respectively). Thus, the approach is promising to develop a strategy for debittering citrus juices by biotransforming limonin at a faster rate.
1 citations
Dissertation•
01 Dec 2009
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Journal Article•
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.
289,852 citations
Journal Article•
TL;DR: An overview of the state-of-the art in the production of the enzyme and its application in the manufacture of 6-aminopenicillanic acid, the key intermediate in semi-synthetic penicillins production is presented.
155 citations
TL;DR: Penicillin amidase was extracted from Escherichia coli ATCC 9637, grown on phenylacetic, acid and glutamate, and purified by fractional ion with streptomycin sulphate, ammonium sulphate and polyethylene glycol, followed by chromatography on DEAE–cellulose.
Abstract: Penicillin amidase was extracted from Escherichia coli ATCC 9637, grown on phenylacetic, acid and glutamate, and purified by fractional ion with streptomycin sulphate, ammonium sulphate and polyethylene glycol, followed by chromatography on DEAE–cellulose. The purification factor was 100–200 × and the overall yield was about 115%. The enzyme was chemically attached to derivatives of cellulose and the kinetics of these insolubilized penicillin amidase preparations was investigated.
78 citations
TL;DR: Mise au point d'un materiel immobilise a activite enzymatique conservee, utilisable pour la production industrielle de penicilline semi-synthetique sur un support type chitosane est decrite.
Abstract: Mise au point d'un materiel immobilise a activite enzymatique conservee, utilisable pour la production industrielle de penicilline semi-synthetique. L'immobilisation de penicilline acylase sur un support type chitosane est decrite: differentes formes du support sont testees: poudre, particules ou billes de chitosane. Differentes methodes d'immobilisation sont egalement presentees: adsorption-reticulation, liaison covalente. Deux types de reacteur sont utilises suivant le type de support choisi: reacteur agite pour la poudre et les particules de chitosane, reacteur lit fixe pour les billes. L'influence de ces differents types d'immobilisation sur l'activite enzymatique de la penicilline G acylase est mesuree
63 citations
01 Sep 1976
TL;DR: The enzyme activity of the immobilized cell column was stable, and its half-life was 17 days at 40°C and 42 days at 30°C, and from the effluent of the column, 6-APA was easily obtained in a good yield.
Abstract: For continuous production of 6-aminopenicillanic acid (6-APA) the microbial cells ofEscherichia coli ATCC 9637 having high penicillin amidase (penicillin amidohydrolase, E.C. 3. 5. 1. 11) activity were immobilized by entrapment in a polyacrylamide gel lattice. Enzymatic properties of penicillin amidase of the immobilizedE. coli cells were investigated and compared with those of the intact cells. With regard to optimal pH and temperature, no marked difference was observed. The heat stability was somewhat increased by immobilization of the cells. The enzyme activity of the immobilized cell column was stable, and its half-life was 17 days at 40°C and 42 days at 30°C. From the effluent of the column, 6-APA was easily obtained in a good yield.
60 citations