Membrane-bound and extracellular β-lactamase production with developmental regulation in Streptomyces griseus NRRL B-2682

• Published in: Microbiology (Society for General Microbiology) Vol. 144; no. 8; pp. 2169 - 2177
• Main Authors: DEAK, E, SZABO, I, KALMACZHELYI, A, GAL, Z, BARABAS, G, PENYIGE, A
• Format: Journal Article
• Language: English
• Published: Reading Society for General Microbiology 01-08-1998
• Subjects: Acetylmuramyl-Alanyl-Isoglutamine - pharmacology; Amino Acid Sequence; Anti-Bacterial Agents - pharmacology; Bacteriology; beta-Lactamases - biosynthesis; beta-Lactamases - isolation & purification; Biological and medical sciences; Extracellular Space - enzymology; Extracellular Space - microbiology; Fundamental and applied biological sciences. Psychology; Membrane Proteins - metabolism; Metabolism. Enzymes; Microbiology; Molecular Sequence Data; Mutation; N-Acetylmuramoyl-L-alanine Amidase - metabolism; Penicillin G - pharmacology; Peptide Fragments - pharmacology; Peptides; Peptidoglycan - pharmacology; Protease Inhibitors - pharmacology; Spores, Bacterial - drug effects; Spores, Bacterial - enzymology; Streptomyces griseus; Streptomyces griseus - drug effects; Streptomyces griseus - enzymology; Streptomyces griseus - physiology; Streptomycetaceae; Enzyme; Biosynthesis; β-Lactamase; Extracellular; Actinomycetales; Characterization; Membrane enzyme; Regulation(control); Sporulation; Bacteria; Hydrolases; Actinomycetes; Streptomyces griseus
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/00221287-144-8-2169

A new type of beta-lactamase has been isolated and characterized in Streptomyces griseus NRRL B-2682. The enzyme has membrane-bound and extracellular forms. Biochemical characterization of some of the properties of the enzyme showed that it belongs to the class A group of penicillinases. Comparison of the membrane-bound and extracellular forms of the beta-lactamases suggests that they seem to be differently processed forms of the same enzyme. The N-terminal amino acid sequence of the extracellular form of the beta-lactamase showed a high degree of similarity to a D-aminopeptidase of another Streptomyces griseus strain. Secretion of the beta-lactamase was affected by the differentiation state of the strain since in spontaneous non-sporulating mutants only the membrane-bound form was present. In accordance with this when sporulation of the wild-type strain was inhibited it failed to secrete extracellular beta-lactamase. Addition of globomycin to the non-sporulating cells liberated the enzyme from the membrane, indicating that the protein is processed normally by signal peptidase II and a glyceride-thioether group, together with a fatty acid amide-linkage, is responsible for the attachment of the enzyme to the cellular membrane. Under sporulation-repressed conditions addition of peptidoglycan fragments and analogues or inhibition of cell wall biosynthesis by penicillin-G induced beta-lactamase secretion and also restored sporulation both in solid and submerged cultures. These results confirm that beta-lactamase secretion is tightly coupled to the sporulation process in S. griseus.