Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth
Abstract
1. Introduction
2. Results and Discussion
2.1. In Silico Analysis of C. difficile and Phages Infecting C. difficile in Search of Lytic Enzymes
2.2. Homology Modelling
2.3. Expression and Lytic Activity of Endolysins
2.4. Characterization of the Catalytic Domains (CWH351—656) of Cell Wall Hydrolase (CWH)
2.5. Host Specificity of Full-Length CWH and CWH351—656
2.6. Optical Microscopy of Lytic Activity
2.7. Effect of CWH351—656 on C. difficile Spore Outgrowth
3. Materials and Methods
3.1. Bacterial Strains and Growth Conditions
3.2. Cloning, Expression, and Purification of CWH and Its Subdomains
3.3. Structural Modelling of CWH351—656
3.4. Lytic Activity Assays
3.5. Optical Microscopy of Lytic Activity
3.6. C. difficile Spore Preparation
3.7. Effect of CWH351—656 on C. difficile Spore Germination and Outgrowth
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Clostridioides Strains | Lysin Activity # | |
---|---|---|
CWH | CWH351—656 | |
Clostridioides difficile 43255 | +++ | +++ |
Clostridioides difficile APC 1401 | ++ | +++ |
Clostridioides difficile 630 | ++ | +++ |
Clostridioides difficile APC 1412 | +++ | +++ |
Clostridioides difficile 28196 | +++ | +++ |
Non-Clostridioides Strains | ||
Bacillus cereus | + | ++ |
Clostridium perfringens | – | – |
Clostridium scindens | + | ++ |
Clostridium symbiosum | – | – |
Enterococcus faecalis | – | + |
Enterococcus faecium | – | – |
Lactobacillus paracasei | – | – |
Lactococcus lactis | – | – |
Listeria innocua | – | + |
Listeria monocytogenes | – | – |
Staphylococcus aureus | – | – |
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Mondal, S.I.; Akter, A.; Draper, L.A.; Ross, R.P.; Hill, C. Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth. Int. J. Mol. Sci. 2021, 22, 5690. https://doi.org/10.3390/ijms22115690
Mondal SI, Akter A, Draper LA, Ross RP, Hill C. Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth. International Journal of Molecular Sciences. 2021; 22(11):5690. https://doi.org/10.3390/ijms22115690
Chicago/Turabian StyleMondal, Shakhinur Islam, Arzuba Akter, Lorraine A. Draper, R. Paul Ross, and Colin Hill. 2021. "Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth" International Journal of Molecular Sciences 22, no. 11: 5690. https://doi.org/10.3390/ijms22115690
APA StyleMondal, S. I., Akter, A., Draper, L. A., Ross, R. P., & Hill, C. (2021). Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth. International Journal of Molecular Sciences, 22(11), 5690. https://doi.org/10.3390/ijms22115690