Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography
The emetic toxin cereulide is a 1.2 kDa dodecadepsipeptide produced by the food pathogen <i>Bacillus cereus</i>. As cereulide poses a serious health risk to humans, sensitive and specific detection, as well as toxin purification and quantification, methods are of utmost importance. Recen...
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MDPI AG
2021-02-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/13/2/115 |
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| author | Eva Maria Kalbhenn Tobias Bauer Timo D. Stark Mandy Knüpfer Gregor Grass Monika Ehling-Schulz |
| author_facet | Eva Maria Kalbhenn Tobias Bauer Timo D. Stark Mandy Knüpfer Gregor Grass Monika Ehling-Schulz |
| author_sort | Eva Maria Kalbhenn |
| collection | DOAJ |
| description | The emetic toxin cereulide is a 1.2 kDa dodecadepsipeptide produced by the food pathogen <i>Bacillus cereus</i>. As cereulide poses a serious health risk to humans, sensitive and specific detection, as well as toxin purification and quantification, methods are of utmost importance. Recently, a stable isotope dilution assay tandem mass spectrometry (SIDA–MS/MS)-based method has been described, and an method for the quantitation of cereulide in foods was established by the International Organization for Standardization (ISO). However, although this SIDA–MS/MS method is highly accurate, the sophisticated high-end MS equipment required for such measurements limits the method’s suitability for microbiological and molecular research. Thus, we aimed to develop a method for cereulide toxin detection and isolation using equipment commonly available in microbiological and biochemical research laboratories. Reproducible detection and relative quantification of cereulide was achieved, employing reversed phase chromatography (RPC). Chromatographic signals were cross validated by ultraperformance liquid chromatography–mass spectrometry (UPLC–MS/MS). The specificity of the RPC method was tested using a test panel of strains that included non-emetic representatives of the <i>B. cereus</i> group, emetic <i>B. cereus</i> strains, and cereulide-deficient isogenic mutants. In summary, the new method represents a robust, economical, and easily accessible research tool that complements existing diagnostics for the detection and quantification of cereulide. |
| first_indexed | 2024-03-09T05:36:24Z |
| format | Article |
| id | doaj.art-36c8cec888244e628eb969dc3b561c74 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T05:36:24Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-36c8cec888244e628eb969dc3b561c742023-12-03T12:28:01ZengMDPI AGToxins2072-66512021-02-0113211510.3390/toxins13020115Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase ChromatographyEva Maria Kalbhenn0Tobias Bauer1Timo D. Stark2Mandy Knüpfer3Gregor Grass4Monika Ehling-Schulz5Functional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, AustriaFunctional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, AustriaChair of Food Chemistry and Molecular Sensory Science, Technical University of Munich, Lise-Meitner-Straße 34, 85354 Freising, GermanyBundeswehr Institute of Microbiology, Neuherbergstraße 11, 80937 Munich, GermanyBundeswehr Institute of Microbiology, Neuherbergstraße 11, 80937 Munich, GermanyFunctional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine Vienna, 1210 Vienna, AustriaThe emetic toxin cereulide is a 1.2 kDa dodecadepsipeptide produced by the food pathogen <i>Bacillus cereus</i>. As cereulide poses a serious health risk to humans, sensitive and specific detection, as well as toxin purification and quantification, methods are of utmost importance. Recently, a stable isotope dilution assay tandem mass spectrometry (SIDA–MS/MS)-based method has been described, and an method for the quantitation of cereulide in foods was established by the International Organization for Standardization (ISO). However, although this SIDA–MS/MS method is highly accurate, the sophisticated high-end MS equipment required for such measurements limits the method’s suitability for microbiological and molecular research. Thus, we aimed to develop a method for cereulide toxin detection and isolation using equipment commonly available in microbiological and biochemical research laboratories. Reproducible detection and relative quantification of cereulide was achieved, employing reversed phase chromatography (RPC). Chromatographic signals were cross validated by ultraperformance liquid chromatography–mass spectrometry (UPLC–MS/MS). The specificity of the RPC method was tested using a test panel of strains that included non-emetic representatives of the <i>B. cereus</i> group, emetic <i>B. cereus</i> strains, and cereulide-deficient isogenic mutants. In summary, the new method represents a robust, economical, and easily accessible research tool that complements existing diagnostics for the detection and quantification of cereulide.https://www.mdpi.com/2072-6651/13/2/115cereulidereversed phase chromatography (RPC)Äkta™ purepeptide quantificationemetic <i>Bacillus cereus</i>toxin purification |
| spellingShingle | Eva Maria Kalbhenn Tobias Bauer Timo D. Stark Mandy Knüpfer Gregor Grass Monika Ehling-Schulz Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography Toxins cereulide reversed phase chromatography (RPC) Äkta™ pure peptide quantification emetic <i>Bacillus cereus</i> toxin purification |
| title | Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography |
| title_full | Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography |
| title_fullStr | Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography |
| title_full_unstemmed | Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography |
| title_short | Detection and Isolation of Emetic <i>Bacillus cereus</i> Toxin Cereulide by Reversed Phase Chromatography |
| title_sort | detection and isolation of emetic i bacillus cereus i toxin cereulide by reversed phase chromatography |
| topic | cereulide reversed phase chromatography (RPC) Äkta™ pure peptide quantification emetic <i>Bacillus cereus</i> toxin purification |
| url | https://www.mdpi.com/2072-6651/13/2/115 |
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