Specific Isolation of Clostridium botulinum Group I Cells by Phage Lysin Cell Wall Binding Domain with the Aid of S-Layer Disruption

• Published in: International journal of molecular sciences Vol. 23; no. 15; p. 8391
• Main Authors: Zhang, Zhen, Douillard, François P., Korkeala, Hannu, Lindström, Miia
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01-08-2022; MDPI
• Subjects: Acids; Bacteria; Binding; Botulinum toxin; Botulism; cell wall binding domain; Cell walls; Clostridium botulinum; diagnostics; Disruption; Domains; Flow cytometry; Glycine; Gram-positive bacteria; Host specificity; Lysins; Magnetic separation; phage lysin; Phages; Proteins; Risk assessment; S-layer; Strains (organisms); Toxins
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23158391

Clostridium botulinum is a notorious pathogen that raises health and food safety concerns by producing the potent botulinum neurotoxin and causing botulism, a potentially fatal neuroparalytic disease in humans and animals. Efficient methods for the identification and isolation of C. botulinum are warranted for laboratory diagnostics of botulism and for food safety risk assessment. The cell wall binding domains (CBD) of phage lysins are recognized by their high specificity and affinity to distinct types of bacteria, which makes them promising for the development of diagnostic tools. We previously identified CBO1751, which is the first antibotulinal phage lysin showing a lytic activity against C. botulinum Group I. In this work, we assessed the host specificity of the CBD of CBO1751 and tested its feasibility as a probe for the specific isolation of C. botulinum Group I strains. We show that the CBO1751 CBD specifically binds to C. botulinum Group I sensu lato (including C. sporogenes) strains. We also demonstrate that some C. botulinum Group I strains possess an S-layer, the disruption of which by an acid glycine treatment is required for efficient binding of the CBO1751 CBD to the cells of these strains. We further developed CBO1751 CBD-based methods using flow cytometry and magnetic separation to specifically isolate viable cells of C. botulinum Group I. These methods present potential for applications in diagnostics and risk assessment in order to control the botulism hazard.