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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #316712

Title: Fusion of a thermophilic phage endolysin to a Clostridium perfringens-specific cell wall binding domain creates an anti-clostridium antimicrobial with improved thermostability

item Swift, Steven
item Seal, Bruce
item Garrish, Johnna
item Oakley, Brian
item Yeh, Hung-Yueh
item HEITT, KELLI - US Department Of Agriculture (USDA)
item WOOLSEY, REBEKAH - University Of Nevada
item SCHEGG, KTHLEEN - University Of Nevada
item LINE, J. ERIC - US Department Of Agriculture (USDA)
item Donovan, David

Submitted to: Evergreen International Phage Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 7/15/2015
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical 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 to the cell wall binding domain from the endolysin of a C. perfringens-specific bacteriophage. The resulting protein lysed C. perfringens in liquid and solid cultures, was most active at pH8, 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 thermostabile chimeric endolysin that could potentially be utilized as a feed additive to control the bacterium during poultry production.