Project Type

Poster

Publication Date

Spring 4-26-2025

Department or Program

Biological Sciences

College

College of Arts & Sciences

Faculty Mentor #1

Dr. Swati Agrawal

Abstract

Bacteriophages present a promising alternative for combating antibiotic-resistant bacteria without disrupting the normal microbiome or harming human cells. The antibacterial properties of lytic bacteriophages are largely attributed to endolysins, enzymes active in the lytic cycle. Endolysins recognize bacterial cell walls and cleave the peptidoglycan matrix, leading to cell wall disruption. Their specificity and effectiveness against bacterial hosts are determined by two functional domains, including the cell wall binding domain. Our research focuses on characterizing Bacillus phage endolysins. We analyzed the protein sequences of endolysins from six Bacillus phages isolated at the University of Mary Washington. Although these phages were isolated using non-pathogenic Bacillus thuringiensis, they have been shown to infect B. cereus and B. anthracis, both of which are significant human pathogens. We confirmed that the phages Atlee, Jabberwock, and Frank share identical endolysin proteins, while Jackrabbit, Cletus, and Hari share a distinct type. To further investigate their lytic properties, we had a private company clone three types of endolysin proteins into a protein expression vector. Preliminary expression results have shown positive expression for all three types of endolysin proteins via western blot. Furthermore, subsequent spot plating has shown effective antimicrobial activities, and we currently aim to compare the lytic activities of the two endolysin proteins to better understand their effectiveness and specificity in targeting bacterial cell walls. In the future, we plan to assess the influence of environmental factors, such as pH and temperature, on their activity and stability, providing insights into their potential applications in combating antibiotic-resistant bacteria.

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