Effect of immobilization on the penicillin binding and reactivity properties of DD-peptidase from Streptomyces R61.
An affinity gel matrix containing an enzyme (DD-peptidase) with specific beta-lactam binding properties was characterized with respect to its binding and reactivity behavior with penicillin. The data show that immobilization of DDP by reaction with the enzymes susceptible amino groups resulted in changes in catalytic activity on a tripeptide substrate, penicillin binding efficiency and pH stability of drug binding. Properties unaffected by immobilization were the drug-enzyme complex stability, binding reaction mechanism, drug selectivity and method of complex desorption. The affinity of DDP for penicillin-G was investigated by surface plasmon resonance. These characteristics were compared with those of the soluble enzyme. Conditions for elution of the bound drug were determined and a method for immobilizing Streptomyces DDP by which its binding site structure is sustained was also evaluated.
Eng
G Y
GY
US Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, PA 19038, USA.
Maxwell
R J
RJ
D002250:Carbon Radioisotopes; D002268:Carboxypeptidases / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D010400:Penicillin G / Q000378:metabolism*; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; D012995:Solubility; D013302:Streptomyces / Q000201:enzymology*; D017186:Titrimetry
D002250:Carbon Radioisotopes; D004800:Enzymes, Immobilized; D002268:Carboxypeptidases; D046929:Serine-Type D-Ala-D-Ala Carboxypeptidase; D010400:Penicillin G