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: Several novel techniques are illustrated in an attempt to convey the most representative and powerful tools that are currently being developed for both immediate and long term applications in Agriculture, Health Food formulation and production, pharmaceuticals and Medicine.
Abstract: Selected applications of novel techniques in Agricultural Biotechnology, Health Food formulations and Medical Biotechnology are being reviewed with the aim of unraveling future developments and policy changes that are likely to open new niches for Biotechnology and prevent the shrinking or closing the existing ones. Amongst the selected novel techniques with applications to both Agricultural and Medical Biotechnology are: immobilized bacterial cells and enzymes, microencapsulation and liposome production, genetic manipulation of microorganisms, development of novel vaccines from plants, epigenomics of mammalian cells and organisms, as well as biocomputational tools for molecular modeling related to disease and Bioinformatics. Both fundamental and applied aspects of the emerging new techniques are being discussed in relation to their anticipated impact on future biotechnology applications together with policy changes that are needed for continued success in both Agricultural and Medical Biotechnology. Several novel techniques are illustrated in an attempt to convey the most representative and powerful tools that are currently being developed for both immediate and long term applications in Agriculture, Health Food formulation and production, pharmaceuticals and Medicine. The research aspects are naturally emphasized in our review as they are key to further developments in Medical and Agricultural Biotechnology.
11 citations
Cites background from "Studies on improved techniques for ..."
...Covalent binding and cross-linking offer better strength than the previous techniques, however, there is an encountered toxicity in the reagents (Chibata 1979; Kolot 1981; Babu and Panda 1991) that are used to produce immobilization....
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TL;DR: Direct activity determination by a flow-through microcalorimetry in the enzyme thermistor system was employed for a fast comparison of (poly)acrylamide gel-entrapped penicillin G acylase preparations and the validity of the results was corroborated by spectrophotometric measurements.
Abstract: Direct activity determination by a flow-through microcalorimetry in the enzyme thermistor system was employed for a fast comparison of (poly)acrylamide gel-entrapped penicillin G acylase preparations. Composition of the pre-polymerization cocktail and both the storage and operational stabilities of optimal gel-entrapped enzyme preparations isolated from the Escherichia coli industrial strain were optimized by this method. The validity of the results was corroborated by spectrophotometric measurements.
10 citations
Dissertation•
01 Jan 2004
TL;DR: In this article, a biocapteur conductimetrique utilisant des cellules algales Chlorella vulgaris immobilisees is proposed, which consiste a optimiser l’immobilisation des cellule algale sur les electrodes ainsi que les protocoles de mesures des activites phosphatases alcalines and cholinesterases.
Abstract: La preservation des ecosystemes aquatiques necessite des outils d’alarme precoce comme les biocapteurs pour une surveillance in situ et en continu. De tels outils peuvent fournir des informations quant a la nature des polluants a condition d’etre concus pour repondre de maniere specifique. Ce travail propose un biocapteur conductimetrique utilisant des cellules algales Chlorella vulgaris immobilisees. Ces microalgues possedent des enzymes membranaires, notamment certaines phosphatase alcalines et cholinesterases dont l’activite peut alors etre facilement mesuree sur cellule entiere sans extraction prealable. Le principal objectif de ce travail a ete la mise au point de la detection de ces activites avec les biocapteurs afin de detecter, dans le milieu aquatique, la presence de composes toxiques inhibiteurs des activites enzymatiques considerees. La premiere etape a donc consiste a optimiser l’immobilisation des cellules algales sur les electrodes ainsi que les protocoles de mesures des activites phosphatases alcalines et cholinesterases. Des essais de toxicite ont ensuite ete realises afin d’etudier les effets de metaux lourds ainsi que de pesticides organophosphores et carbamates sur les activites phosphatase alcaline et cholinesterase. Ces biocapteurs permettent ainsi la detection de Cd2+ et Zn2+ avec une limite de detection proche de 10ppb. Les premieres experimentations realisees avec le methyl-paraoxon ont montre, pour leur part, que celui-ci inhibe significativement l’activite cholinesterase a la difference du carbofuran et du methyl-parathion.
8 citations
TL;DR: Agar was the suitable matrix to immobilize Bacillus sp.
Abstract: Aims: To investigate the effect of cell immobilization on amylase production by the moderately halophilic bacterium, Bacillus sp strain TSCVKK and to compare the properties of the amylase produced under immobilized conditions with the enzyme produced by the free cells Study Design: Cell immobilization Place and Duration of Study: Department of Chemistry, Biochemistry Lab, Indian Institute of Technology (IIT Madras), Chennai, Tamil Nadu, between Jan 2009 and March 2009 Methodology: Bacillus sp strain TSCVKK was immobilized in alginate, agar, polyacrylamide and gelatin Production of amylase was determined using 3, 5dinitrosalicylic acid (DNS) Effect of NaCl, pH, temperature on the activity of amylase was determined and compared with the amylase produced by the free cells Results: Maximum production of 832 mU/ml was achieved with an initial cell load of 12% (w/v; wet weight) of 24 h grown cells immobilized in 2% agar of 4 mm block size using GSL-2 medium containing 10% NaCl and 15% dextrin at pH 80 at 30oC after 36 h of growth Amylase production was lower when the cells were immobilized in alginate (211 mU/ml) or with the free cells of same biomass concentration as used for immobilization Research Article British Microbiology Research Journal, 2(3): 146-157, 2012 147 (333 mU/ml) Amylase was not produced when gelatin or polyacrylamide was used as the immobilization matrix The immobilized cells in 2% agar could be used up to 5 cycles without much reduction in amylase production Amylase produced through cell immobilization retained all the properties that were shown by amylase produced under submerged fermentation Conclusion: Agar was the suitable matrix to immobilize Bacillus sp strain TSCVKK for amylase production Amylase produced under immobilization conditions retained its temperature, salt and pH requirements Immobilized cells were used for 5 cycles without much decrease in production
8 citations
Dissertation•
01 Jan 2010
TL;DR: In this article, the authors propose le developpement de biocapteurs for the detection of certain families de polluants, i.e., metaux lourds and phosphates.
Abstract: Le controle de la qualite des ecosystemes aquatiques necessite des outils de detection en continu et in situ comme les biocapteurs. Ce travail propose le developpement de biocapteurs pour la detection de certaines familles de polluants. Ces outils sont bases sur la mesure de deux activites enzymatiques de cellules algales representatives des eaux douces et salines. La premiere etape a consiste au choix des activites en fonction des polluants etudies en bioessais. Ainsi nous avons montre que l'activite phosphatase alcaline (APA) est sensible a la presence de metaux lourds et de phosphates et que l'activite esterase (AE) est sensible a la presence de pesticides. Ces resultats nous ont permis de developper differents biocapteurs. Pour la detection des metaux lourds, deux biocapteurs conductimetriques ont ete developpes. Le premier, base sur la mesure de l'APA de la microalgue d'eau douce Chlorella vulgaris (Cv) immobilisee par des monocouches autoassemblees, a permis d'ameliorer la repetabilite et la reproductibilite des mesures. Le deuxieme, base sur la meme mesure, a permis de s'affranchir des effets du phosphate sur l'APA de Cv. Pour la detection du phosphate, un biocapteur conductimetrique a cellules algales a ete developpe, la limite de detection est de 0. 4µM d'ions phosphate. Enfin pour la detection des pesticides en milieu marin, un biocapteur conductimetrique base sur la mesure de l'AE de deux algues marines a ete developpe. Les resultats ont montre qu'il est sensible a la presence du diuron et du glyphosate. Les biocapteurs developpes lors de cette etude, et apres essais sur des echantillons naturels, servirons aux gestionnaires pour une prise de decision.
8 citations
References
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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