Production and characterization of a phage endolysin with putative anti-microbial activity against Cutibacterium acnes
Faculty Mentor
Luis Matos
Document Type
Oral Presentation
Start Date
10-5-2023 2:05 PM
End Date
10-5-2023 2:25 PM
Location
PUB 317
Department
Biology
Abstract
Acne vulgaris (acne) is the eighth most common skin disorder worldwide. Because of the heterogeneous pathology of acne and the increasing antibiotic resistance of the causal agent (Cutibacterium acnes), novel therapeutics need to be developed to treat acne. One option is endolysins, highly conserved enzymes from bacteriophages that disrupt the bacterial cell wall. Endolysins retain bactericidal and bacteriostatic activity when applied to bacteria in vitro and are safe for topical application. We hypothesize that the endolysin (ENDL) from the P100.1 C. acnes bacteriophage will exhibit bacteriostatic activity against C. acnes in vitro. In this study, the ENDL gene was cloned into and expressed by Escherichia coli as a fusion protein to green fluorescent protein (GFP). The region responsible for binding to the bacterial cell wall (CBD) was expressed separately (by E. coli) as a fusion protein to GFP to visualize its binding activity to C. acnes. Expression of the recombinant proteins was successful, given that cultures induced for protein expression were fluorescent. Different lysis methods were tested, including bead beating, bead vortexing, and protein extraction buffer, to optimize yields of the fusion proteins by quantifying relative fluorescence output in the cell lysate. Next, we will compare the growth rates of C. acnes treated with ENDL-GFP to non-treatment controls to evaluate bactericidal effects. Finally, the off-target bactericidal effects of ENDL will be determined. This study aims to characterize the P100.1 endolysin’s structure and activity to evaluate its potential as a novel anti-bacterial acne therapy.
Recommended Citation
Collins, Kathryn, "Production and characterization of a phage endolysin with putative anti-microbial activity against Cutibacterium acnes" (2023). 2023 Symposium. 11.
https://dc.ewu.edu/srcw_2023/res_2023/os1_2023/11
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Production and characterization of a phage endolysin with putative anti-microbial activity against Cutibacterium acnes
PUB 317
Acne vulgaris (acne) is the eighth most common skin disorder worldwide. Because of the heterogeneous pathology of acne and the increasing antibiotic resistance of the causal agent (Cutibacterium acnes), novel therapeutics need to be developed to treat acne. One option is endolysins, highly conserved enzymes from bacteriophages that disrupt the bacterial cell wall. Endolysins retain bactericidal and bacteriostatic activity when applied to bacteria in vitro and are safe for topical application. We hypothesize that the endolysin (ENDL) from the P100.1 C. acnes bacteriophage will exhibit bacteriostatic activity against C. acnes in vitro. In this study, the ENDL gene was cloned into and expressed by Escherichia coli as a fusion protein to green fluorescent protein (GFP). The region responsible for binding to the bacterial cell wall (CBD) was expressed separately (by E. coli) as a fusion protein to GFP to visualize its binding activity to C. acnes. Expression of the recombinant proteins was successful, given that cultures induced for protein expression were fluorescent. Different lysis methods were tested, including bead beating, bead vortexing, and protein extraction buffer, to optimize yields of the fusion proteins by quantifying relative fluorescence output in the cell lysate. Next, we will compare the growth rates of C. acnes treated with ENDL-GFP to non-treatment controls to evaluate bactericidal effects. Finally, the off-target bactericidal effects of ENDL will be determined. This study aims to characterize the P100.1 endolysin’s structure and activity to evaluate its potential as a novel anti-bacterial acne therapy.
