Journal
MOLECULAR MICROBIOLOGY
Volume 91, Issue 6, Pages 1136-1147Publisher
WILEY
DOI: 10.1111/mmi.12523
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Funding
- Medical Research Council
- Maplethorpe Trust
- National Institute for Health Research University College London Hospitals Biomedical Research Centre
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A mixed culture of Pseudomonas fluorescens and Pusillimonas noertemanii, obtained by soil enrichment, elaborated an enzyme (EnvD) which rapidly hydrolysed poly--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 -links in both d- and l-glutamic acid-containing polymers. The stability at 37 degrees C was markedly superior to that of CapD, a -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.anthracisPasteur within 5min. We conclude from this in vitro study that rEnvD shows promise as a potential therapeutic for the treatment of anthrax.
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