Purification and properties of a beta-lactamase from Alcaligenes dentrificans subsp. xylosoxydans.
A penicillin beta-lactamase was purified from a strain of Alcaligenes dentrificans subsp. xylosoxydans resistant to beta-lactam antibiotics. The purified enzyme preparation gave a single protein band on polyacrylamide gel electrophoresis, and its molecular weight was 18,000 from sodium dodecylsulphate-acrylamide gel electrophoresis and gel filtration. Its isoelectric point was 9.8, the optimal pH was 8.5 and the optimal temperature was 35 degrees C. The enzyme hydrolyzed penicillin G and ampicillin more rapidly than cephalosporins. Relative rates, with penicillin G as 100, were: ampicillin, 102; carbenicillin, 15; cloxacillin, less than 1; piperacillin, 9; cephaloridine, 41; cefoperazone, 36; cefpiramide, 36 and cefmenoxime, 14. Clavulanic acid, sulbactam, imipenem, and cephamycins had low affinities for the enzyme. The enzyme activity was inhibited by iodine, Hg2+, clavulanic acid and sulbactam.
D000421:Alcaligenes / Q000201:enzymology*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003470:Culture Media; D004790:Enzyme Induction; D006868:Hydrolysis; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010405:Penicillinase / Q000302:isolation & purification; D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases / Q000302:isolation & purification / Q000378:metabolism*
D003470:Culture Media; D065093:beta-Lactamase Inhibitors; D010405:Penicillinase; D001618:beta-Lactamases