Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.5.1.4 (deaminase)
 5,113 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

D-(-)-Phenylglycyl-beta-lactamide amidohydrolase was isolated from Xanthomonas sp., purified, and characterized. A characteristic feature of the enzyme is its high specificity for substrates containing an alpha-aminophenylacetic group in the acyl moiety. Cephalexin and D-C-(-)-phenylglycine methyl ester (MEPG), being nonspecific penicillin acylase (EC 3.5.1.11) substrates, have the highest values of bimolecular constant kcat/Km (2.8 x 10(5) and 2.0 x 10(5) M-1 x s-1, respectively) in the case of amidohydrolase. On the contrary, benzylpenicillin is not hydrolyzed by D-(-)-phenylglycyl-beta-lactamide amidohydrolase. The other peculiarity of the enzyme is its affinity for the charged forms of substrates. Using the amidohydrolase, it was found that the values of delta Go'pH7.0 for hydrolysis of the amide bond in cephalexin and ampicillin are -3.3 and -2.3 kJ mol-1, respectively. They are less by a minimum of 2.7 kJ mol-1 than those for other beta-lactam antibiotics. Detailed thermodynamic and kinetic studies of the synthesis of cephalexin from MEPG and 7-aminodeacetoxycephalosporanic acid (7-ADCA) catalyzed by D-(-)-phenylglycyl-beta-lactamide amidohydrolase were undertaken. A kinetic scheme is proposed which describes well the experimental curves. The value of conversion of "nucleus" was found to be 76% when the synthesis was carried out from a 31.5 mM solution of 7-ADCA and an 88.5 mM solution of MEPG at pH 6.2 (optimum conditions). A 75% conversion of 7-aminocephalosporanic acid (7-ACA) was achieved in the synthesis of cephaloglycine catalyzed by D-(-)-phenylglycyl-beta-lactamide amidohydrolase.
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PMID:Synthesis of beta-lactam antibiotics containing alpha-aminophenylacetyl group in the acyl moiety catalyzed by D-(-)-phenylglycyl-beta-lactamide amidohydrolase. 776 56

A new hydrophobic and catalytic membrane was prepared by immobilizing Penicillin G acylase (PGA, EC.3.5.1.11) from E. coli on a nylon membrane, chemically grafted with butylmethacrylate (BMA). Hexamethylenediamine (HMDA) and glutaraldehyde (Glu) were used as a spacer and coupling agent, respectively. PGA was used for the enzymatic synthesis of cephalexin, using D(-)-phenylglycine methyl ester (PGME) and 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) as substrates. Several factors affecting this reaction, such as pH, temperature, and concentrations of substrates were investigated. The results indicated good enzyme-binding efficiency of the pre-treated membrane, and an increased stability of the immobilized PGA towards pH and temperature. Calculation of the activation energies showed that cephalexin production by the immobilized biocatalyst was limited by diffusion, resulting in a decrease of enzyme activity and substrate affinity. Temperature gradients were employed as a way to reduce the effects of diffusion limitation. Cephalexin was found to linearly increase with the applied temperature gradient. A temperature difference of about 3 degrees C across the catalytic membrane resulted into a cephalexin synthesis increase of 100% with a 50% reduction of the production times. The advantage of using non-isothermal bioreactors in biotechnological processes, including pharmaceutical applications, is also discussed.
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PMID:Advantages of using non-isothermal bioreactors for the enzymatic synthesis of antibiotics: the penicillin G acylase as enzyme model. 1211 22

The effect of methanol on the kinetically controlled synthesis of cephalexin by free and immobilized penicillin G acylase (PGA) was investigated. Catalytic and hydrophobic membranes were obtained by chemical grafting, activation, and PGA immobilization on hydrophobic nylon supports. Butyl methacrylate (BMA) was used as graft monomer. Increasing concentrations of methanol were found to cause a greater deleterious effect on the activity of free than on that of the immobilized enzyme. Methanol, however, improved the kinetic stability of cephalexin synthesized by free PGA, resulting in higher maximum yields. By contrast, immobilized PGA reached 100% yields even in the absence of the cosolvent. Cephalexin synthesis by the catalytic membrane was also performed in a non-isothermal bioreactor. Under these conditions, a 94% increase of the synthetic activity and complete conversion of the limiting substrate to cephalexin were obtained. The addition of methanol reduced the non-isothermal activity increase. The physical cause responsible for the non-isothermal behavior of the hydrophobic catalytic membrane was identified in the process of thermodialysis.
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PMID:Enzyme reaction engineering: effect of methanol on the synthesis of antibiotics catalyzed by immobilized penicillin G acylase under isothermal and non-isothermal conditions. 1236 48