Authors: Jonathan E Schmitz Maria Cristina Ossiprandi Kareem R Rumah Vincent A Fischetti
Publish Date: 2010/11/18
Volume: 89, Issue: 6, Pages: 1783-1795
Abstract
With their ability to lyse Grampositive bacteria phage lytic enzymes or lysins have received a great deal of attention as novel antiinfective agents The number of known genes encoding these peptidoglycan hydrolases has increased markedly in recent years due in large part to advances in DNA sequencing technology As the genomes of more and more bacterial species/strains are sequenced lysinencoding open reading frames ORFs can be readily identified in lysogenized prophage regions In the current study we sought to assess lysin diversity for the medically relevant pathogen Clostridium perfringens The sequenced genomes of nine C perfringens strains were computationally mined for prophage lysins and lysinlike ORFs revealing several dozen proteins of various enzymatic classes Of these lysins a muramidase from strain ATCC 13124 termed PlyCM was chosen for recombinant analysis based on its dissimilarity to previously characterized C perfringens lysins Following expression and purification various biochemical properties of PlyCM were determined in vitro including pH/saltdependence and temperature stability The enzyme exhibited activity at low μg/ml concentrations a typical value for phage lysins It was active against 23 of 24 strains of C perfringens tested with virtually no activity against other clostridial or nonclostridial species Overall PlyCM shows potential for development as an enzybiotic agent demonstrating how expanding genomic databases can serve as rich pools for biotechnologically relevant proteinsThis research was funded by NIH/NIAID grants AI057472 and AI11822 to VAF JES acknowledges the kind support of the NIH MSTP program Weill Cornell/Rockefeller/SloanKettering grant GM 07739 The authors would like to thank Dr Davise Larone of New York Hospital for providing clostridial strains We also acknowledge Prof Ezio Bottarelli of the University of Parma for his valued insights as well as Ms Cinzia Reverberi and Mr Roberto Lurisi for technical assistance Electron micrographs were acquired by Ms Eleana Sphicas of the Rockefeller University Electron Microscopy Core Facility
Keywords: