Bioengineering of bacteriophage-derived endolysin against Clostridioides difficile
1
Issued Date
2022
Copyright Date
2022
Resource Type
Language
eng
File Type
application/pdf
Access Rights
open access
Rights
ผลงานนี้เป็นลิขสิทธิ์ของมหาวิทยาลัยมหิดล ขอสงวนไว้สำหรับเพื่อการศึกษาเท่านั้น ต้องอ้างอิงแหล่งที่มา ห้ามดัดแปลงเนื้อหา และห้ามนำไปใช้เพื่อการค้า
Rights Holder(s)
Mahidol University
Suggested Citation
Wichuda Phothichaisri (2022). Bioengineering of bacteriophage-derived endolysin against Clostridioides difficile. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/114205
Title
Bioengineering of bacteriophage-derived endolysin against Clostridioides difficile
Author(s)
Abstract
Clostridioides difficile is a significant cause of antibiotic-associateddiarrhea and pseudomembranous colitis. The extensive use of antibiotics drives the emergence of new antibiotic-resistant C. difficile strains with high virulence and low susceptibility to current antibiotic treatment. Therefore, the development of alternativetherapy is required. Phage endolysin is a peptidoglycan hydrolase enzyme potentially an antibacterial agent for bacterial infection treatment. To apply endolysin fortherapeutic purposes, the fundamental role of the enzyme is needed. In this study, an identical endolysin was identified from two C. difficile phages, ΦHN16-1 and ΦHN50, designated as CD16/50. The sequence analysis revealed a modular architecture of an N-terminal enzymatically active domain (EAD) and a C-terminal cell-wall binding domain(CBD). In vitro characterization confirmed that the EAD possessed bacteriolytic activitywhile the CBD bound bacterial cell-wall polysaccharide. The EAD alone exhibited lyticactivity faster than the full-length, suggesting a CBD-independent activity. The studyalso showed that the CBD formed a homodimer essential for interaction with cell-wallpolymer. Interestingly, the hidden Markov model analysis suggested that the CBD islikely derived from the CWB2 motif of C. difficile cell-wall proteins but exhibits ahigher binding affinity to bacterial cell-wall polysaccharides. Finally, endolysindiffusion and sequential cytolytic assays suggested that CBD is required for theendolysin to be trapped into post-lytic bacterial remnants, implying its physiologicalroles in limiting enzyme diffusion, preserving neighboring host cells, and allowing thephage progeny to initiate new rounds of infection. Collectively, this study showed the role of CBD in endolysin regulation, which may provide an insight into designing potentendolysins against C. difficile.
Degree Name
Doctor of Philosophy
Degree Level
Doctoral degree
Degree Department
Faculty of Science
Degree Discipline
Biochemistry
Degree Grantor(s)
Mahidol University