KINETIC STUDIES OF PENICILLIN G HYDROLYSIS WITH IMMOBILIZED PENICILLIN ACYLASE IN AN ELECTRODIALYZER

1996 ◽  
Vol 147 (1) ◽  
pp. 99-117 ◽  
Author(s):  
TING-CHIA HUANG ◽  
DONG-HWANG CHEN ◽  
SUNG-SHYONG WANG
Keyword(s):  
Kinetic Studies ◽  
Penicillin Acylase ◽  
Penicillin G ◽  
Immobilized Penicillin Acylase

scholarly journals Immobilization of penicillin acylase from Escherichia coli on commercial sepabeads EC-EP carrier

2007 ◽  
pp. 173-182 ◽  
Author(s):  
Milena Zuza ◽  
Slavica Siler-Marinkovic ◽  
Zorica Knezevic
Keyword(s):  
Escherichia Coli ◽  
Conformational Stability ◽  
Maximum Yield ◽  
Penicillin Acylase ◽  
Covalent Immobilization ◽  
Penicillin G ◽  
Kinetic Properties ◽  
Enzyme Coupling ◽  
Immobilized Penicillin Acylase ◽  
Coupling Yield

This paper describes the covalent immobilization of penicillin G acylase from Escherichia coli on sepabeads EC-EP, an epoxy-activated polymethacrylic carrier and kinetic properties of the immobilized enzyme. The selected enzyme belongs to a class of biocatalysts whose industrial interest is due to their versatility to mediate hydrolysis of penicillins and semi-synthetic ?-lactam antibiotics synthesis reactions. About 2.7 mg of the pure enzyme was immobilized onto each gram of sepabeads with an enzyme coupling yield of 96.9%. However, it seems that the activity coupling yield is not correlated with the amount of enzyme bound and the maximum yield of 89.4% can be achieved working at low enzyme loading (0.14 mg g-1). Immobilization of the penicillin acylase resulted in slightly different pH activity profile and temperature optima, indicating that the immobilization by this method imparted structural and conformational stability of this enzyme. It appears that both free and immobilized penicillin acylase followed simple Michaelis-Menten kinetics, implying the same reaction mechanism in both systems.

scholarly journals The use of immobilized penicillin acylase for estimation of penicillin G.

1993 ◽  
Vol 40 (1) ◽  
pp. 93-94
Author(s):  
A Marciniak-Rusek ◽  
L Paśś-Dziegielewska
Keyword(s):  
Penicillin Acylase ◽  
Penicillin G ◽  
Immobilized Penicillin Acylase

Optimization of 6-aminopenicillanic acid (6-APA) production by using a new immobilized penicillin acylase

2000 ◽  
Vol 32 (3) ◽  
pp. 173 ◽  
Author(s):  
Jesús Torres-Bacete ◽  
Miguel Arroyo ◽  
Raquel Torres-Guzmán ◽  
Isabel de la Mata ◽  
María Pilar Castillón ◽  
...  
Keyword(s):  
Penicillin Acylase ◽  
Immobilized Penicillin Acylase

Low temperature effect on production of ampicillin and cephalexin in ethylene glycol medium with immobilized penicillin acylase

2006 ◽  
Vol 41 (9) ◽  
pp. 1924-1931 ◽  
Author(s):  
Carolina Aguirre ◽  
Paola Opazo ◽  
Mineri Venegas ◽  
Ruby Riveros ◽  
Andrés Illanes
Keyword(s):  
Low Temperature ◽  
Ethylene Glycol ◽  
Temperature Effect ◽  
Penicillin Acylase ◽  
Immobilized Penicillin Acylase

scholarly journals Inactivation of penicillin acylase from Kluyvera citrophila by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline: a case of time-dependent non-covalent enzyme inhibition

1993 ◽  
Vol 291 (3) ◽  
pp. 907-914 ◽  
Author(s):  
J Martín ◽  
J M Mancheño ◽  
R Arche
Keyword(s):  
Enzyme Assay ◽  
Ph Dependence ◽  
Penicillin Acylase ◽  
Enzyme Inactivation ◽  
Time Dependent ◽  
Penicillin G ◽  
Reversible Binding ◽  
Ph Increase ◽  
Kinetics Of ◽  
Covalent Activation

Penicillin acylase (PA) from Kluyvera citrophila was inhibited by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), a specific carboxy-group-reactive reagent. Enzyme activity progressively decreased to a residual value depending on EEDQ concentration. Neither enzymic nor non-enzymic decomposition of EEDQ is concomitant with PA inactivation. Moreover, enzyme re-activation is achieved by chromatographic removal of EEDQ, pH increase or displacement of the reagent with penicillin G. It was then concluded that PA inactivation is due to an equilibrium reaction. The kinetics of enzyme inactivation was analysed by fitting data to theoretical equations derived in accordance with this mechanism. Corrections for re-activation during the enzyme assay were a necessary introduction. The pH-dependence of the rate constant for EEDQ hydrolysis either alone or in the presence of enzyme was studied by u.v. spectroscopy. It turned out to be coincident with the pH-dependence of the forward and reverse rate constants for the inactivation process. It is suggested that previous protonation of the EEDQ molecule is required for these reactions to occur. The thermodynamic values associated with the overall reaction showed little change. Finally it is proposed that the inactivation of PA by EEDQ proceeds through a two-step reaction. The initial and rapid reversible binding is followed by a slow, time-dependent, non-covalent, reversible inactivating step. The expected behaviour in the case of enzyme modification by covalent activation of carboxy residues is also reviewed.

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