Viruses 2015, 7(6), 3019-3034; https://doi.org/10.3390/v7062758
A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability
1
Animal 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, USA
2
Poultry 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, USA
3
Nevada Proteomics Center, University of Nevada School of Medicine, Manville Medical Building, 1664 N. Virginia Street, Reno, NV 89557-MS0330, USA
†
Current address: Biology Program, Oregon State University Cascades, Bend, OR 97701, USA.
‡
Current address: Western University College of Veterinary Medicine, Pomona, CA 91766, USA.
*
Author to whom correspondence should be addressed.
Academic Editors: Abram Aertsen and Rob Lavigne
Received: 30 April 2015 / Revised: 20 May 2015 / Accepted: 9 June 2015 / Published: 12 June 2015
(This article belongs to the Special Issue 21st Evergreen International Phage Biology Meeting - Emerging Research)
Abstract
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. View Full-TextKeywords:
alternative antimicrobial; bacteriophage; endolysin; food safety; feed additive; peptidoglycan hydrolase; thermostabile; thermostable
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MDPI and ACS Style
Swift, S.M.; Seal, B.S.; Garrish, J.K.; Oakley, B.B.; Hiett, K.; Yeh, H.-Y.; Woolsey, R.; Schegg, K.M.; Line, J.E.; Donovan, D.M. A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability. Viruses 2015, 7, 3019-3034.