Purification of a soil bacteria exotoxin using silkworm toxicity to measure specific activity

Kimihito Usui, Shinya Miyazaki, Chikara Kaito, Kazuhisa Sekimizu

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Overnight cultures of 16 of 25 species of soil bacterium injected into silkworm hemolymph killed the silkworms. Injection of culture supernatants of five of these 16 bacterial species into the hemolymph also killed silkworms. Analysis of 16S rRNA indicated that the toxic species were Bacillus species. We purified an exotoxin produced by one of the Bacillus species by measuring the specific activity based on silkworm toxicity. The final step of purification revealed that the silkworm-killing activity was due to a 34-kDa protein. LC/MS/MS analysis revealed that the 34-kDa protein is sphingomyelinase C of Bacillus cereus. We propose that the silkworm infection model is useful for identifying pathogenic bacteria from soil and purifying bacterial toxins.

Original languageEnglish
Pages (from-to)59-62
Number of pages4
JournalMicrobial Pathogenesis
Volume46
Issue number2
DOIs
Publication statusPublished - Feb 1 2009
Externally publishedYes

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Keywords

  • Purification
  • Silkworm
  • Soil bacteria
  • Sphingomyelinase

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

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