Characterization of four prophage endolysins specific for clostridium perfringens

US10544406B2 · kind B2 · utility

Assignees

• The United States of America, as represented by the Secretary of Agriculture · US
• WESTERN UNIVERSITY OF HEALTH SCIENCES · US
• UNIVERSITY OF MARYLAND, COLLEGE PARK · US

Inventors

• David M. Donovan · Baltimore, US
• Jerel Waters · Randallstown, US
• Dayana T. Rowley · Beltsville, US
• Steven M. Swift · Sumner, US
• Brian B. Oakley · Claremont, US

Key dates

• Filing date: Jan 5, 2018
• Grant date: Jan 28, 2020
• Expiry date: Jun 9, 2038

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y302/01
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Clostridium perfringens can cause food poisoning and is a major agent in necrotic enteritis. As laws banning the use of antibiotics in animal feed become more common, the need for alternatives to antibiotics becomes greater. Peptidoglycan hydrolases that target the cell wall of specific bacteria are one such alternative. Genes for four endolysins, PlyCP10, PlyCP18, PlyCP33, and PlyCP41, were found within clusters of phage associated genes, likely prophages from strains Cp10, Cp18, Cp33, and Cp41. PlyCP18 and PlyCP33 harbor L-alanine amidase catalytic domains, and PlyCP10 and PlyCP41 have glycosyl hydrolase catalytic domains as predicted by BlastP and PFAM searches. All four genes were synthesized with E. coli codon optimization, expressed in E. coli expression vectors with a 6×His tag for nickel column purification, and the recombinant proteins purified. The four endolysins were capable of lysing the 66 C. perfringens strains tested but not the other bacteria tested.