title: A poly-γ-(D)-glutamic acid depolymerase that degrades the protective capsule of Bacillus anthracis.
abstract: A mixed culture of Pseudomonas fluorescens and Pusillimonas noertemanii, obtained by soil enrichment, elaborated an enzyme (EnvD) which rapidly hydrolysed poly-γ-d-glutamic acid (PDGA), the constituent of the anti-phagocytic capsule conferring virulence on Bacillus anthracis. The EnvD gene is carried on the P. noertemanii genome but co-culture is required for the elaboration of PDGA depolymerase activity. EnvD showed strong sequence homology to dienelactone hydrolases from other Gram-negative bacteria, possessed no general protease activity but cleaved γ-links in both d- and l-glutamic acid-containing polymers. The stability at 37°C was markedly superior to that of CapD, a γ-glutamyltranspeptidase with PDGA depolymerase activity. Recombinant EnvD was recovered from inclusion bodies in soluble form from an Escherichia coli expression vector and the enzyme stripped the PDGA capsule from the surface of B. anthracis Pasteur within 5 min. We conclude from this in vitro study that rEnvD shows promise as a potential therapeutic for the treatment of anthrax.
authors.affiliation: University College London School of Pharmacy, 29-39 Brunswick Square, London, WC1N 1AX, UK.
authors.forename: David
authors.initials: D
authors.lastname: Negus
authors.forename: Peter W
authors.initials: PW
authors.lastname: Taylor
mesh_terms: D001408:Bacillus anthracis / Q000737:chemistry*; D016667:Bacterial Capsules / Q000378:metabolism*; D001711:Biotransformation; D004795:Enzyme Stability; D006867:Hydrolases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011099:Polyglutamic Acid / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012988:Soil Microbiology; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D011099:Polyglutamic Acid; D006867:Hydrolases