Hydrolysis of penicillin G by combination of immobilized penicillin acylase and electrodialysis

1992 ◽  
Vol 39 (2) ◽  
pp. 171-175 ◽  
Author(s):  
Fumihiro Ishimura ◽  
Ken-Ichi Suga
Keyword(s):  
Penicillin Acylase ◽  
Penicillin G ◽  
Immobilized Penicillin Acylase ◽  
Hydrolysis Of

scholarly journals Immobilization of alginate-PAC on Sepabeads EC-HA support

2011 ◽  
Vol 65 (4) ◽  
pp. 431-437 ◽  
Author(s):  
Milena Zuza ◽  
Nenad Milosavic ◽  
Zorica Knezevic-Jugovic
Keyword(s):  
Phenylacetic Acid ◽  
Immobilized Enzyme ◽  
Penicillin Acylase ◽  
Covalent Immobilization ◽  
Enzyme Inactivation ◽  
Penicillin G ◽  
Ph Profile ◽  
Covalently Linked ◽  
Hydrolysis Of ◽  
Derivatives Of

Penicillin acylase (PAC) is an important industrial enzyme for the production of many ?-lactam antibiotics. It is capable of catalyzing the hydrolysis of penicillin G (Pen G) to generate phenylacetic acid (PAA) and 6-aminopenicillanic acid (6-APA). In this paper, in order to prevent enzyme inactivation, an attempt of coupling enzyme modification and immobilization was presented. Chemical modification was promoted to introduce carbohydrate moiety into the PAC molecule, capable of being covalently linked to an amino support. This seems to provide a possibility to couple the enzyme without risking a reaction at the active site which might cause a loss of activity. PAC molecules were modified by cross-linking with polyaldehyde derivatives of alginate in order to add them new and useful functions. Immobilization of alginate-PAC on Sepabeads EC-HA was used as a model system in order to demonstrate the potential of this strategy. Optimal conditions for covalent immobilization of alginate-PAC from Escherichia coli on support Sepabeads EC-HA, were investigated. The immobilized enzyme was then characterized by evaluating the potential effects of immobilization on its thermal stability, temperature and pH profile in comparison with native non-modified PAC and modified non-immobilized PAC. The maximum amount of the alginate-PAC coupled on the dry support of 99 mg/g was satisfactory. Deactivation rate constants at 50 ?C for free PAC, alginate-PAC and alginate-PAC immobilized on Sepabeads EC-HA were 2,32; 50,65 and 1,68 h-1, respectively. Alginate-PAC and alginate-PAC immobilized on Sepabeads EC-HA had the same pH and temperature optimum as the native non-modified PAC.

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

scholarly journals The kinetics of acylation and deacylation of penicillin acylase from Escherichia coli ATCC 11105: evidence for lowered pKa values of groups near the catalytic centre

1999 ◽  
Vol 338 (1) ◽  
pp. 235-239 ◽  
Author(s):  
Manuel MORILLAS ◽  
Martin L. GOBLE ◽  
Richard VIRDEN
Keyword(s):  
Rate Constant ◽  
Conformational Transition ◽  
Ph Dependence ◽  
Penicillin Acylase ◽  
Penicillin G ◽  
Pka Values ◽  
Energy Compensation ◽  
Guanidinium Group ◽  
Kinetics Of ◽  
Hydrolysis Of

Penicillin G acylase catalysed the hydrolysis of 4-nitrophenyl acetate with a kcat of 0.8 s-1 and a Km of 10 µM at pH 7.5 and 20 °C. Results from stopped-flow experiments fitted a dissociation constant of 0.16 mM for the Michaelis complex, formation of an acetyl enzyme with a rate constant of 32 s-1 and a subsequent deacylation step with a rate constant of 0.81 s-1. Non-linear Van't Hoff and Arrhenius plots for these parameters, measured at pH 7.5, may be partly explained by a conformational transition affecting catalytic groups, but a linear Arrhenius plot for the ratio of the rate constant for acylation relative to KS was consistent with energy-compensation between the binding of the substrate and catalysis of the formation of the transition state. At 20 °C, the pH-dependence of kcat was similar to that of kcat/Km, indicating that formation of the acyl-enzyme did not affect the pKa values (6.5 and 9.0) of an acidic and basic group in the active enzyme. The heats of ionization deduced from values of pKa for kcat, which measures the rate of deacylation, are consistent with α-amino and guanidinium groups whose pKa values are decreased in a non-polar environment. It is proposed that, for catalytic activity, the α-amino group of the catalytic SerB1 and the guanidinium group of ArgB263 are required in neutral and protonated states respectively.

scholarly journals THE HYDROLYSIS OF PENICILLIN G TO 6-AMINO PENICILLANIC ACID BY ENTRAPPED PENICILLIN ACYLASE

1973 ◽  
Vol 26 (4) ◽  
pp. 228-232 ◽  
Author(s):  
W. MARCONI ◽  
F. CECERE ◽  
F. MORISI ◽  
G. DELLA PENNA ◽  
B. RAPPUOLI
Keyword(s):  
Penicillin Acylase ◽  
Penicillin G ◽  
Penicillanic Acid ◽  
Hydrolysis Of

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
Sign in / Sign up

Export Citation Format

Share Document