A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability
Clostridium perfringens is the third leading cause of human foodborne bacterial disease and is the presumptive etiologic agent of necrotic enteritis among chickens. Treatment of poultry with antibiotics is becoming less acceptable. Endolysin enzymes are potential replacements for antibiotics. Man...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2015-06-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1999-4915/7/6/2758 |
| _version_ | 1818991769918898176 |
|---|---|
| author | Steven M. Swift Bruce S. Seal Johnna K. Garrish Brian B. Oakley Kelli Hiett Hung-Yueh Yeh Rebekah Woolsey Kathleen M. Schegg John Eric Line David M. Donovan |
| author_facet | Steven M. Swift Bruce S. Seal Johnna K. Garrish Brian B. Oakley Kelli Hiett Hung-Yueh Yeh Rebekah Woolsey Kathleen M. Schegg John Eric Line David M. Donovan |
| author_sort | Steven M. Swift |
| collection | DOAJ |
| description | Clostridium perfringens is the third leading cause of human foodborne bacterial disease and is the presumptive etiologic agent of necrotic enteritis among chickens. Treatment of poultry with antibiotics is becoming less acceptable. Endolysin enzymes are potential replacements for antibiotics. Many enzymes are added to animal feed during production and are subjected to high-heat stress during feed processing. To produce a thermostabile endolysin for treating poultry, an E. coli codon-optimized gene was synthesized that fused the N-acetylmuramoyl-L-alanine amidase domain from the endolysin of the thermophilic bacteriophage ɸGVE2 to the cell-wall binding domain (CWB) from the endolysin of the C. perfringens-specific bacteriophage ɸCP26F. The resulting protein, PlyGVE2CpCWB, lysed C. perfringens in liquid and solid cultures. PlyGVE2CpCWB was most active at pH 8, had peak activity at 10 mM NaCl, 40% activity at 150 mM NaCl and was still 16% active at 600 mM NaCl. The protein was able to withstand temperatures up to 50° C and still lyse C. perfringens. Herein, we report the construction and characterization of a thermostable chimeric endolysin that could potentially be utilized as a feed additive to control the bacterium during poultry production. |
| first_indexed | 2024-12-20T20:15:32Z |
| format | Article |
| id | doaj.art-dd191b64d19d46e1aa70523afbeebc84 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-20T20:15:32Z |
| publishDate | 2015-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-dd191b64d19d46e1aa70523afbeebc842022-12-21T19:27:44ZengMDPI AGViruses1999-49152015-06-01763019303410.3390/v7062758v7062758A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved ThermostabilitySteven M. Swift0Bruce S. Seal1Johnna K. Garrish2Brian B. Oakley3Kelli Hiett4Hung-Yueh Yeh5Rebekah Woolsey6Kathleen M. Schegg7John Eric Line8David M. Donovan9Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 10300 Baltimore Avenue, Bldg. 230, BARC-East, Beltsville, MD 20705, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USANevada Proteomics Center, University of Nevada School of Medicine, Manville Medical Building, 1664 N. Virginia Street, Reno, NV 89557-MS0330, USANevada Proteomics Center, University of Nevada School of Medicine, Manville Medical Building, 1664 N. Virginia Street, Reno, NV 89557-MS0330, USAPoultry Microbiological Safety and Processing Research Unit, Richard B. Russell Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 950 College Station Road, Athens, GA 30605, USAAnimal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, 10300 Baltimore Avenue, Bldg. 230, BARC-East, Beltsville, MD 20705, USAClostridium perfringens is the third leading cause of human foodborne bacterial disease and is the presumptive etiologic agent of necrotic enteritis among chickens. Treatment of poultry with antibiotics is becoming less acceptable. Endolysin enzymes are potential replacements for antibiotics. Many enzymes are added to animal feed during production and are subjected to high-heat stress during feed processing. To produce a thermostabile endolysin for treating poultry, an E. coli codon-optimized gene was synthesized that fused the N-acetylmuramoyl-L-alanine amidase domain from the endolysin of the thermophilic bacteriophage ɸGVE2 to the cell-wall binding domain (CWB) from the endolysin of the C. perfringens-specific bacteriophage ɸCP26F. The resulting protein, PlyGVE2CpCWB, lysed C. perfringens in liquid and solid cultures. PlyGVE2CpCWB was most active at pH 8, had peak activity at 10 mM NaCl, 40% activity at 150 mM NaCl and was still 16% active at 600 mM NaCl. The protein was able to withstand temperatures up to 50° C and still lyse C. perfringens. Herein, we report the construction and characterization of a thermostable chimeric endolysin that could potentially be utilized as a feed additive to control the bacterium during poultry production.http://www.mdpi.com/1999-4915/7/6/2758alternative antimicrobialbacteriophageendolysinfood safetyfeed additivepeptidoglycan hydrolasethermostabilethermostable |
| spellingShingle | Steven M. Swift Bruce S. Seal Johnna K. Garrish Brian B. Oakley Kelli Hiett Hung-Yueh Yeh Rebekah Woolsey Kathleen M. Schegg John Eric Line David M. Donovan A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability Viruses alternative antimicrobial bacteriophage endolysin food safety feed additive peptidoglycan hydrolase thermostabile thermostable |
| title | A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability |
| title_full | A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability |
| title_fullStr | A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability |
| title_full_unstemmed | A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability |
| title_short | A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability |
| title_sort | thermophilic phage endolysin fusion to a clostridium perfringens specific cell wall binding domain creates an anti clostridium antimicrobial with improved thermostability |
| topic | alternative antimicrobial bacteriophage endolysin food safety feed additive peptidoglycan hydrolase thermostabile thermostable |
| url | http://www.mdpi.com/1999-4915/7/6/2758 |
| work_keys_str_mv | AT stevenmswift athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT brucesseal athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT johnnakgarrish athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT brianboakley athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT kellihiett athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT hungyuehyeh athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT rebekahwoolsey athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT kathleenmschegg athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT johnericline athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT davidmdonovan athermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT stevenmswift thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT brucesseal thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT johnnakgarrish thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT brianboakley thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT kellihiett thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT hungyuehyeh thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT rebekahwoolsey thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT kathleenmschegg thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT johnericline thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability AT davidmdonovan thermophilicphageendolysinfusiontoaclostridiumperfringensspecificcellwallbindingdomaincreatesananticlostridiumantimicrobialwithimprovedthermostability |