A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability
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Authors
Swift, Steven M.
Seal, Bruce S.
Garrish, Johnna K.
Oakley, Brian B.
Hiett, Kelli
Yeh, Hung-Yueh
Woolsey, Rebekah
Schegg, Kathleen M.
Line, John E.
Donovan, David M.
Issue Date
2015
Type
Article
Language
Keywords
alternative antimicrobial , bacteriophage , endolysin , food safety , feed additive , peptidoglycan hydrolase , thermostabile , thermostable
Alternative Title
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 phi GVE2 to the cell-wall binding domain (CWB) from the endolysin of the C. perfringens-specific bacteriophage phi CP26F. 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 degrees 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.
Description
Citation
Swift, S., Seal, B., Garrish, J., Oakley, B., Hiett, K., Yeh, H.-Y., … Donovan, D. (2015). A Thermophilic Phage Endolysin Fusion to a Clostridium perfringens-Specific Cell Wall Binding Domain Creates an Anti-Clostridium Antimicrobial with Improved Thermostability. Viruses, 7(6), 3019�"3034. doi:10.3390/v7062758
Publisher
License
Creative Commons Attribution 4.0 International
Journal
Volume
Issue
PubMed ID
DOI
ISSN
1999-4915