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A novel and highly specific phage endolysin cell wall binding domain for detection of Bacillus cereus

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Abstract

Rapid, specific and sensitive detection of pathogenic bacteria is crucial for public health and safety. Bacillus cereus is harmful as it causes foodborne illness and a number of systemic and local infections. We report a novel phage endolysin cell wall-binding domain (CBD) for B. cereus and the development of a highly specific and sensitive surface plasmon resonance (SPR)-based B. cereus detection method using the CBD. The newly discovered CBD from endolysin of PBC1, a B. cereus-specific bacteriophage, provides high specificity and binding capacity to B. cereus. By using the CBD-modified SPR chips, B. cereus can be detected at the range of 105–108 CFU/ml. More importantly, the detection limit can be improved to 102 CFU/ml by using a subtractive inhibition assay based on the pre-incubation of B. cereus and CBDs, removal of CBD-bound B. cereus, and SPR detection of the unbound CBDs. The present study suggests that the small and genetically engineered CBDs can be promising biological probes for B. cereus. We anticipate that the CBD-based SPR-sensing methods will be useful for the sensitive, selective, and rapid detection of B. cereus.

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Acknowledgments

This research was supported by the Public Welfare & Safety research program (NRF-2012M3A2A1051684, NRF-2012M3A2A1051682) through the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning (MSIP), Global Frontier Project (H-GUARD_2013M3A6B2078950, H-GUARD_2014M3A6B2060489) through the Center for BioNano Health-Guard funded by the MSIP, and KRIBB initiative Research Program.

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    Authors and Affiliations

    1. Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Center for Food and Bioconvergence, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-742, Korea

      Minsuk Kong & Sangryeol Ryu

    2. BioNanotechnology Research Center, KRIBB, Daejeon, 305-806, Korea

      Jieun Sim, Taejoon Kang, Hoang Hiep Nguyen & Bong Hyun Chung

    3. BioNano Health Guard Research Center, KRIBB, Daejeon, 305-806, Korea

      Jieun Sim, Taejoon Kang & Bong Hyun Chung

    4. Major of Nanobiotechnology, Korea University of Science and Technology, Daejeon, 305-806, Korea

      Jieun Sim, Taejoon Kang, Hoang Hiep Nguyen & Bong Hyun Chung

    5. MicoBioMed Ltd, Daejeon, 305-806, Korea

      Hyun Kyu Park

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    1. Minsuk Kong
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    Correspondence to Bong Hyun Chung or Sangryeol Ryu.

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    M. Kong and J. Sim contributed equally to this work.

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    Kong, M., Sim, J., Kang, T. et al. A novel and highly specific phage endolysin cell wall binding domain for detection of Bacillus cereus . Eur Biophys J 44, 437–446 (2015). https://doi.org/10.1007/s00249-015-1044-7

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