Abstract
Clostridioides difficile is a spore-forming enteric pathogen causing life-threatening diarrhoea and colitis. Microbial disruption caused by antibiotics has been linked with susceptibility to, and transmission and relapse of, C. difficile infection. Therefore, there is an urgent need for novel therapeutics that are effective in preventing C. difficile growth, spore germination, and outgrowth. In recent years bacteriophage-derived endolysins and their derivatives show promise as a novel class of antibacterial agents. In this study, we recombinantly expressed and characterized a cell wall hydrolase (CWH) lysin from C. difficile phage, phiMMP01. The full-length CWH displayed lytic activity against selected C. difficile strains. However, removing the N-terminal cell wall binding domain, creating CWH351—656, resulted in increased and/or an expanded lytic spectrum of activity. C. difficile specificity was retained versus commensal clostridia and other bacterial species. As expected, the putative cell wall binding domain, CWH1—350, was completely inactive. We also observe the effect of CWH351—656 on preventing C. difficile spore outgrowth. Our results suggest that CWH351—656 has therapeutic potential as an antimicrobial agent against C. difficile infection.
Highlights
Accepted: 24 May 2021Clostridioides difficile is a Gram-positive anaerobic spore-former that is the causative agent of toxin-mediated colitis in humans [1]
We found a cell wall hydrolase (CWH) lysin encoded by the C. difficile phage phiMMP01 that is unique in domain architecture compared to other characterized
BLAST searches of the protein data bank (PDB) showed some identity to putative cell wall hydrolases encoded by a Tn916-like element in C. difficile 630 (4HPE), NlpC/P60 family protein of Bacillus cereus (3H41) and Trichomonas vaginalis (6BIQ), invasion protein (3PBI), and peptidoglycan endopeptidase (4Q4G) of Mycobacterium tuberculosis
Summary
Clostridioides difficile (formerly known as Clostridium difficile) is a Gram-positive anaerobic spore-former that is the causative agent of toxin-mediated colitis in humans [1]. C. difficile infection (CDI) has become a worldwide health threat that has a high incidence in both hospital and community settings, with increased morbidity and mortality [2]. It has recently been reported that there are nearly 462,100 CDI cases annually in the United States, leading to at least 12,800 fatalities [3,4]. The estimated annual cost of healthcare associated with C. difficile infection in the USA is $5 billion and €3 billion in Europe [5,6]. Disruptions to the gut microbiome, most often because of protracted antibiotic therapy, allows C. difficile spore germination, growth, and toxin production in the colon [7]
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