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ARS Home » Research » Publications at this Location » Publication #199969
Title: Designer Antimicrobials: Peptidoglycan Hydrolase Module Shuffling

Author
item Donovan, David
item DONG, S - UNIVERSITY OF ALABAMA
item Garrett, Wesley
item Foster Frey, Juli
item ROUSSEAU, G - UNIVERSITE LAVAL, QUEBEC
item MOINEAU, S - UNIVERSITE LAVAL, QUEBE
item PRITCHARD, D - UNIVERSITY OF ALABAMA

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: 2/16/2006
Publication Date: 2/16/2006
Citation: Donovan, D.M., Dong, S., Garrett, W.M., Foster Frey, J.A., Rousseau, G.M., Moineau, S., Pritchard, D.G. 2006. Designer Antimicrobials: Peptidoglycan Hydrolase Module Shuffling. American Society for Microbiology.

Interpretive Summary:

Technical Abstract: Bovine mastitis (mammary gland infections) exists on every dairy farm, affects 1/3 of the animals, and costs the US dairy industry about $2 Billion annually. There are multiple mastitis-causing pathogens but streptococci and staphylococci are the most predominant. Broad spectrum antibiotics are often less than 50% effective, with culling often necessary. Transgenic cattle resistant to S. aureus mastitis challenge have been created via mammary gland expression of a peptidoglycan (PG) hydrolase (lysostaphin) that is lytic for S. aureus cell walls (Nat. Biotechnol. 23:445-51). Phage endolysins are a prodigious source of novel PG hydrolases that often contain two discrete enzymatic domains that each target a unique bond in the PG. We have performed deletion analysis and site-directed mutagenesis to show that the streptococcal phage B30 endolysin Acm glycosidase (lysozyme-like) domain, although highly conserved and enzymatically active on cell wall preparations, does not contribute significantly to cell lysis when added externally in turbidity reduction assays. However, the N-terminal B30 CHAP endopeptidase domain is lytic toward multiple streptococci in the absence of the Acm domain (and C-terminal SH3b domain). Fusions of PG hydrolase domains have been shown to maintain their domain-specific hydrolytic activities. Fusing either the highly active B30 CHAP endopeptidase domain or the full length B30 endolysin to full length lysostaphin creates an antimicrobial that is active against both S. aureus and several streptococci. This designer antimicrobial approach demonstrates the facility of constructing highly specific antibacterial proteins that target multiple unrelated pathogens simultaneously.