Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Minor Coat Protein pIII Domain (N1N2) of Bacteriophage CTXф Confers a Novel Surface Plasmon Resonance Biosensor for Rapid Detection of Vibrio cholerae

  • Shin, Hae Ja (Department of Bio-Pharmaceutical Engineering, College of Bio-Health, Dongseo University) ;
  • Hyeon, Seok Hywan (Department of Molecular Biology, College of Natural Sciences, Pusan National University) ;
  • Cho, Jae Ho (Department of Molecular Biology, College of Natural Sciences, Pusan National University) ;
  • Lim, Woon Ki (Department of Molecular Biology, College of Natural Sciences, Pusan National University)
  • Received : 2021.09.26
  • Accepted : 2021.11.10
  • Published : 2021.12.28
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Abstract

Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 103 to 109 CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.

Keywords

Acknowledgement

This work was supported by a grant from the Basic Science Research Program (2016R1D1A1B01015961) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Republic of Korea.

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