Identifying Phage Lytic Enzymes: Past, Present, and Future

Schmitz, Jonathan E.; Schuch, Raymond; Fischetti, Vincent A.

doi:10.1002/9780470570548.ch10

Cited by 1 publication

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“…Most lysins characterized to date were cloned through traditional phage genomic techniques (Schmitz et al 2010). Here, a phage is isolated through environmental sampling or prophage induction, and its genomic DNA is extracted following laboratory culture.…”

Section: Introductionmentioning

confidence: 99%

Lytic enzyme discovery through multigenomic sequence analysis in Clostridium perfringens

Schmitz

Ossiprandi

Rumah

et al. 2010

Appl Microbiol Biotechnol

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With their ability to lyse Gram-positive bacteria, phage lytic enzymes (or lysins) have received a great deal of attention as novel anti-infective 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, lysin-encoding 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 lysin-like 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/salt-dependence 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 proteins.

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Section: Introductionmentioning

confidence: 99%