Extraction, purification and characterization of a peptidoglycan hydrolase from vegetative cells of Clostridium perfringens type A.

Abstract

A peptidoglycan hydrolase (PGH), which causes the germination and lysis of the coat-stripped spores of Clostridium perfringens, was extracted from perfringens vegetative cells. The enzyme was partially purified and showed an MW of 125, 000 and a pI of 7.6. It was sensitive to a variety of monovalent and divalent cations and was inhibited by sulfhydryl-active agents. The sites of PGH action, the cell wall and the spore-cortex, were determined by the release of free amino groups from cell wall fragments and the release of soluble fluorescent products from fluorescamine-labelled cortical fragments of the C. perfringens spores. The enzyme appears distinct from the extracellular initiation protein produced by C. perfringens. PGH may be an autolysin with corticolytic activity.