Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents
ABSTRACT As antibiotic resistance has become a major global threat, the World Health Organization (WHO) has urgently called for alternative strategies for control of bacterial infections. Endolysin, a phage-encoded protein, can degrade bacterial peptidoglycan (PG) and disrupt bacterial growth. Accor...
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| Language: | English |
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American Society for Microbiology
2025-01-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01170-24 |
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| author | Jindanuch Maneekul Amanda Chiaha Rachel Hughes Faith Labry Joshua Saito Matthew Almendares Brenda N. Banda Leslie Lopez Nyeomi McGaskey Melizza Miranda Jenil Rana Brandon R. Zadeh Lee E. Hughes |
| author_facet | Jindanuch Maneekul Amanda Chiaha Rachel Hughes Faith Labry Joshua Saito Matthew Almendares Brenda N. Banda Leslie Lopez Nyeomi McGaskey Melizza Miranda Jenil Rana Brandon R. Zadeh Lee E. Hughes |
| author_sort | Jindanuch Maneekul |
| collection | DOAJ |
| description | ABSTRACT As antibiotic resistance has become a major global threat, the World Health Organization (WHO) has urgently called for alternative strategies for control of bacterial infections. Endolysin, a phage-encoded protein, can degrade bacterial peptidoglycan (PG) and disrupt bacterial growth. According to the WHO, there are only three endolysin products currently in clinical phase development. In this study, we explore novel endolysins from Streptomyces phages as only a few of them have been experimentally characterized. Using several bioinformatics tools, we identified nine different functional domain combinations from 250 Streptomyces phages putative endolysins. LazerLemon gp35 (CHAP; LL35lys), Nabi gp26 (amidase; Nb26lys), and Tribute gp42 (PGRP/amidase; Tb42lys) were selected for experimental studies. We hypothesized that (i) the proteins of interest will have the ability to degrade purified PG, and (ii) the proteins will have potential antimicrobial activity against bacteria from families of importance in antibiotic resistance, such as ESKAPE safe relatives (Enterococcus raffinosus, Staphylococcus epidermidis, Klebsiella aerogenes, Acinetobacter baylyi, Pseudomonas putida, and Escherichia coli). LL35lys, Nb26lys, and Tb42lys exhibit PG-degrading activity on zymography and hydrolysis assay. The enzymes (100 µg/mL) can reduce PG turbidity to 32%–40%. The killing assay suggests that Tb42lys has a broader range (E. coli, P. putida, A. baylyi and K. aerogenes). While Nb26lys better attacks Gram-negative than -positive bacteria, LL35lys can only reduce the growth of the Gram-positive ESKAPE strains but does so effectively with a low MIC90 of 2 µg/mL. A higher concentration (≥300 µg/mL) of Nb26lys is needed to inhibit P. putida and K. aerogenes. From 250 putative endolysins, bioinformatic methods were used to select three putative endolysins for cloning and study: LL35lys, Nb26lys, and Tb42lys. All have shown PG-degrading activity, a critical function of endolysin. With a low MIC, LL35lys shows activity for the Gram-positive ESKAPE strains, while Nb26lys and Tb42lys are active against the Gram negatives. Therefore, endolysins from Streptomyces phages have potential as possible antimicrobial agents against ESKAPE bacteria.IMPORTANCEAs antibiotic resistance has become a major global threat, the World Health Organization (WHO) has urgently called for alternative strategies for control of bacterial infections. Endolysin, a phage-encoded protein, can degrade bacterial peptidoglycan in the bacterial cell wall and disrupt bacterial growth. According to the WHO, there are only three endolysin products currently in clinical phase development. In this study we explored novel endolysins from Streptomyces phages as only a few of them have been experimentally characterized. Using several bioinformatics tools, we identified nine different combinations of functional enzymatic domain types from 250 Streptomyces bacteriophages possible endolysins. From these, three potential endolysins were selected for experimental characterization. All three showed positive results in degrading cell wall material and disrupting bacterial growth, indicating their potential as possible antimicrobial agents. |
| format | Article |
| id | doaj-art-cffdcf42e6714b028c8b77ea7fbc6558 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-cffdcf42e6714b028c8b77ea7fbc65582025-01-07T14:05:18ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-01-0113110.1128/spectrum.01170-24Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agentsJindanuch Maneekul0Amanda Chiaha1Rachel Hughes2Faith Labry3Joshua Saito4Matthew Almendares5Brenda N. Banda6Leslie Lopez7Nyeomi McGaskey8Melizza Miranda9Jenil Rana10Brandon R. Zadeh11Lee E. Hughes12Department of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USADepartment of Biological Sciences, University of North Texas, Denton, Texas, USAABSTRACT As antibiotic resistance has become a major global threat, the World Health Organization (WHO) has urgently called for alternative strategies for control of bacterial infections. Endolysin, a phage-encoded protein, can degrade bacterial peptidoglycan (PG) and disrupt bacterial growth. According to the WHO, there are only three endolysin products currently in clinical phase development. In this study, we explore novel endolysins from Streptomyces phages as only a few of them have been experimentally characterized. Using several bioinformatics tools, we identified nine different functional domain combinations from 250 Streptomyces phages putative endolysins. LazerLemon gp35 (CHAP; LL35lys), Nabi gp26 (amidase; Nb26lys), and Tribute gp42 (PGRP/amidase; Tb42lys) were selected for experimental studies. We hypothesized that (i) the proteins of interest will have the ability to degrade purified PG, and (ii) the proteins will have potential antimicrobial activity against bacteria from families of importance in antibiotic resistance, such as ESKAPE safe relatives (Enterococcus raffinosus, Staphylococcus epidermidis, Klebsiella aerogenes, Acinetobacter baylyi, Pseudomonas putida, and Escherichia coli). LL35lys, Nb26lys, and Tb42lys exhibit PG-degrading activity on zymography and hydrolysis assay. The enzymes (100 µg/mL) can reduce PG turbidity to 32%–40%. The killing assay suggests that Tb42lys has a broader range (E. coli, P. putida, A. baylyi and K. aerogenes). While Nb26lys better attacks Gram-negative than -positive bacteria, LL35lys can only reduce the growth of the Gram-positive ESKAPE strains but does so effectively with a low MIC90 of 2 µg/mL. A higher concentration (≥300 µg/mL) of Nb26lys is needed to inhibit P. putida and K. aerogenes. From 250 putative endolysins, bioinformatic methods were used to select three putative endolysins for cloning and study: LL35lys, Nb26lys, and Tb42lys. All have shown PG-degrading activity, a critical function of endolysin. With a low MIC, LL35lys shows activity for the Gram-positive ESKAPE strains, while Nb26lys and Tb42lys are active against the Gram negatives. Therefore, endolysins from Streptomyces phages have potential as possible antimicrobial agents against ESKAPE bacteria.IMPORTANCEAs antibiotic resistance has become a major global threat, the World Health Organization (WHO) has urgently called for alternative strategies for control of bacterial infections. Endolysin, a phage-encoded protein, can degrade bacterial peptidoglycan in the bacterial cell wall and disrupt bacterial growth. According to the WHO, there are only three endolysin products currently in clinical phase development. In this study we explored novel endolysins from Streptomyces phages as only a few of them have been experimentally characterized. Using several bioinformatics tools, we identified nine different combinations of functional enzymatic domain types from 250 Streptomyces bacteriophages possible endolysins. From these, three potential endolysins were selected for experimental characterization. All three showed positive results in degrading cell wall material and disrupting bacterial growth, indicating their potential as possible antimicrobial agents.https://journals.asm.org/doi/10.1128/spectrum.01170-24endolysinStreptomyces phagebioinformaticsantimicrobial activity |
| spellingShingle | Jindanuch Maneekul Amanda Chiaha Rachel Hughes Faith Labry Joshua Saito Matthew Almendares Brenda N. Banda Leslie Lopez Nyeomi McGaskey Melizza Miranda Jenil Rana Brandon R. Zadeh Lee E. Hughes Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents Microbiology Spectrum endolysin Streptomyces phage bioinformatics antimicrobial activity |
| title | Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents |
| title_full | Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents |
| title_fullStr | Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents |
| title_full_unstemmed | Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents |
| title_short | Investigating novel Streptomyces bacteriophage endolysins as potential antimicrobial agents |
| title_sort | investigating novel streptomyces bacteriophage endolysins as potential antimicrobial agents |
| topic | endolysin Streptomyces phage bioinformatics antimicrobial activity |
| url | https://journals.asm.org/doi/10.1128/spectrum.01170-24 |
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