Purification and properties of inducible penicillin beta-lactamase isolated from Pseudomonas maltophilia

Two types of beta-lactamase were found in the cell-free extract from Pseudomonas maltophilia GN12873. One was an inducible penicillin beta-lactamase, and the other was an inducible cephalosporin beta-lactamase. The purified penicillin beta-lactamase gave a single protein band on polyacrylamide gel electrophoresis. The isoelectric point was 6.9, and the approximate molecular weight was 118,000 by gel filtration and 26,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that this enzyme consisted of four subunits. For the hydrolysis of penicillin G, the optimal pH was 8.0 and the optimal temperature was 35 degrees C. The enzyme activity was inhibited by cephamycin derivatives, carpetimycins A and B, iodine, and HgCl2, but not by clavulanic acid. Furthermore, beta-lactamase activity was almost completely inhibited by EDTA but was recovered by the addition of zinc ion. The enzyme showed a unique substrate profile, hydrolyzing N-formimidoyl thienamycin at a significant rate.

[1]  S. Mitsuhashi,et al.  Antimicrobial and beta-lactamase inhibitory activities of carpetimycins A and B, new carbapenem antibiotics , 1982, Antimicrobial Agents and Chemotherapy.

[2]  S. Mitsuhashi,et al.  Beta-lactamase stability and antibacterial activity of cefmenoxime (SCE-1365), a novel cephalosporin , 1981, Antimicrobial Agents and Chemotherapy.

[3]  H. Nakazawa,et al.  Effect of N-formimidoyl thienamycin (MK0787) on beta-lactamases and activity against beta-lactamase-producing strains , 1980, Antimicrobial Agents and Chemotherapy.

[4]  S. Mitsuhashi,et al.  Purification and Properties of a New β-Lactamase from Pseudomonas cepacia , 1980, Antimicrobial Agents and Chemotherapy.

[5]  S. Mitsuhashi,et al.  Purification and properties of cephalosporinase in Escherichia coli , 1980, Antimicrobial Agents and Chemotherapy.

[6]  T. Hashizume,et al.  Antibacterial activities of a new stabilized thienamycin, N-formimidoyl thienamycin, in comparison with other antibiotics , 1980, Antimicrobial Agents and Chemotherapy.

[7]  V. Yu,et al.  Susceptibility of Pseudomonas maltophilia to antimicrobial agents, singly and in combination , 1979, Antimicrobial Agents and Chemotherapy.

[8]  A. J. Markovetz,et al.  Resistance plasmids in Pseudomonas cepacia 4G9 , 1979, Journal of bacteriology.

[9]  R. Fass,et al.  Effect of Divalent Cation Concentrations on the Antibiotic Susceptibilities of Nonfermenters Other than Pseudomonas aeruginosa , 1979, Antimicrobial Agents and Chemotherapy.

[10]  S. Lapage,et al.  Distribution in clinical material and identification of Pseudomonas maltophilia. , 1979, Journal of clinical pathology.

[11]  R. H. Olsen,et al.  Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5 , 1978, Journal of bacteriology.

[12]  J. D. Semel,et al.  Pseudomonas maltophilia pseudosepticemia. , 1978, The American journal of medicine.

[13]  T. Sawai,et al.  Inducible Oxacillin-Hydrolyzing Penicillinase in Aeromonas hydrophila Isolated from Fish , 1976, Antimicrobial Agents and Chemotherapy.

[14]  R. Sykes,et al.  The -lactamases of Gram-negative bacteria and their rle in resistance to -lactam antibiotics , 1976 .

[15]  J. W. Dale,et al.  The dimeric nature of an R-factor mediated β-lactamase , 1976 .

[16]  N. Terakado,et al.  Biochemical Properties of a Penicillin Beta-Lactamase Mediated by R Factor from Bordetella bronchiseptica , 1975, Antimicrobial Agents and Chemotherapy.

[17]  U. K. Laemmli,et al.  Maturation of the head of bacteriophage T4. I. DNA packaging events. , 1973, Journal of molecular biology.

[18]  J. Fischer.  Pseudomonas maltophilia endocarditis after replacement of the mitral valve: a case study. , 1973, The Journal of infectious diseases.

[19]  C. O'callaghan,et al.  Comparison of assay techniques for β-lactamase activity , 1973 .

[20]  M. Swartz,et al.  Nonfermentative gram-negative bacilli of nosocomial interest. , 1970, The American journal of medicine.

[21]  B. Davis.  DISC ELECTROPHORESIS – II METHOD AND APPLICATION TO HUMAN SERUM PROTEINS * , 1964, Annals of the New York Academy of Sciences.

[22]  P. Andrews,et al.  Estimation of the molecular weights of proteins by Sephadex gel-filtration. , 1964, The Biochemical journal.

[23]  Oliver H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[24]  R. Sykes,et al.  The β-Lactamases of Gram-Negative Bacteria and their Possible Physiological Role , 1973 .

[25]  G. Gilardi.  Pseudomonas maltophilia infections in man. , 1969, American journal of clinical pathology.

[26]  E. Abraham,et al.  Zinc as a cofactor for cephalosporinase from Bacillus cereus 569. , 1966, The Biochemical journal.

[27]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .