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

Kimihito Usui; Shinya Miyazaki; Chikara Kaito; Kazuhisa Sekimizu

doi:10.1016/j.micpath.2008.10.010

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 journal › Article › peer-review

17 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 language	English
Pages (from-to)	59-62
Number of pages	4
Journal	Microbial Pathogenesis
Volume	46
Issue number	2
DOIs	https://doi.org/10.1016/j.micpath.2008.10.010
Publication status	Published - Feb 2009
Externally published	Yes

Keywords

Purification
Silkworm
Soil bacteria
Sphingomyelinase

ASJC Scopus subject areas

Microbiology
Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.micpath.2008.10.010

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title = "Purification of a soil bacteria exotoxin using silkworm toxicity to measure specific activity",

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.",

keywords = "Purification, Silkworm, Soil bacteria, Sphingomyelinase",

author = "Kimihito Usui and Shinya Miyazaki and Chikara Kaito and Kazuhisa Sekimizu",

note = "Funding Information: This work was supported by Grants-in-Aid for Scientific Research (19041022, 20790057, 20390021). This study was supported in part by the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO) and Genome Pharmaceuticals Institute.",

year = "2009",

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doi = "10.1016/j.micpath.2008.10.010",

language = "English",

volume = "46",

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journal = "Microbial Pathogenesis",

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AU - Usui, Kimihito

AU - Miyazaki, Shinya

AU - Kaito, Chikara

AU - Sekimizu, Kazuhisa

N1 - Funding Information: This work was supported by Grants-in-Aid for Scientific Research (19041022, 20790057, 20390021). This study was supported in part by the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO) and Genome Pharmaceuticals Institute.

PY - 2009/2

Y1 - 2009/2

N2 - 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.

AB - 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.

KW - Purification

KW - Silkworm

KW - Soil bacteria

KW - Sphingomyelinase

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SN - 0882-4010

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

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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