title: Evolutionarily distinct bacteriophage endolysins featuring conserved peptidoglycan cleavage sites protect mice from MRSA infection.
abstract: OBJECTIVE
  In the light of increasing drug resistance in Staphylococcus aureus, bacteriophage endolysins [peptidoglycan hydrolases (PGHs)] have been suggested as promising antimicrobial agents. The aim of this study was to determine the antimicrobial activity of nine enzymes representing unique homology groups within a diverse class of staphylococcal PGHs.
  
  
  METHODS
  PGHs were recombinantly expressed, purified and tested for staphylolytic activity in multiple in vitro assays (zymogram, turbidity reduction assay and plate lysis) and against a comprehensive set of strains (S. aureus and CoNS). PGH cut sites in the staphylococcal peptidoglycan were determined by biochemical assays (Park-Johnson and Ghuysen procedures) and MS analysis. The enzymes were tested for their ability to eradicate static S. aureus biofilms and compared for their efficacy against systemic MRSA infection in a mouse model.
  
  
  RESULTS
  Despite similar modular architectures and unexpectedly conserved cleavage sites in the peptidoglycan (conferred by evolutionarily divergent catalytic domains), the enzymes displayed varying degrees of in vitro lytic activity against numerous staphylococcal strains, including cell surface mutants and drug-resistant strains, and proved effective against static biofilms. In a mouse model of systemic MRSA infection, six PGHs provided 100% protection from death, with animals being free of clinical signs at the end of the experiment.
  
  
  CONCLUSIONS
  Our results corroborate the high potential of PGHs for treatment of S. aureus infections and reveal unique antimicrobial and biochemical properties of the different enzymes, suggesting a high diversity of potential applications despite highly conserved peptidoglycan target sites.
authors: Animal Biosciences and Biotechnology Laboratory, ANRI, NEA, ARS, USDA, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland mathias.schmelcher@hest.ethz.ch.
authors: Mathias
authors: M
authors: Schmelcher
authors: Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland Institute for Bioscience and Biotechnology Research, University of Maryland, 9600 Gudelsky Drive, Rockville, MD 20850, USA.
authors: Yang
authors: Y
authors: Shen
authors: Institute for Bioscience and Biotechnology Research, University of Maryland, 9600 Gudelsky Drive, Rockville, MD 20850, USA Department of Veterinary Medicine, University of Maryland, 8075 Greenmead Drive, College Park, MD 20742, USA.
authors: Daniel C
authors: DC
authors: Nelson
authors: Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland.
authors: Marcel R
authors: MR
authors: Eugster
authors: Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland.
authors: Fritz
authors: F
authors: Eichenseher
authors: Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland.
authors: Daniela C
authors: DC
authors: Hanke
authors: Institute of Food, Nutrition and Health, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland.
authors: Martin J
authors: MJ
authors: Loessner
authors: Department of Biochemistry and Molecular Genetics, MCLM 552, University of Alabama at Birmingham, 1530 3rd Ave., Birmingham, AL 35294-0005, USA.
authors: Shengli
authors: S
authors: Dong
authors: Department of Biochemistry and Molecular Genetics, MCLM 552, University of Alabama at Birmingham, 1530 3rd Ave., Birmingham, AL 35294-0005, USA.
authors: David G
authors: DG
authors: Pritchard
authors: Channing Laboratory, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors: Jean C
authors: JC
authors: Lee
authors: Animal Biosciences and Biotechnology Laboratory, ANRI, NEA, ARS, USDA, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA.
authors: Stephen C
authors: SC
authors: Becker
authors: Animal Biosciences and Biotechnology Laboratory, ANRI, NEA, ARS, USDA, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA.
authors: Juli
authors: J
authors: Foster-Frey
authors: Animal Biosciences and Biotechnology Laboratory, ANRI, NEA, ARS, USDA, 10300 Baltimore Ave., Beltsville, MD 20705-2350, USA.
authors: David M
authors: DM
authors: Donovan
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D016470:Bacteremia / Q000188:drug therapy / Q000382:microbiology; D001435:Bacteriophages / Q000201:enzymology*; D001691:Biological Therapy / Q000379:methods*; D002473:Cell Wall / Q000187:drug effects; D004195:Disease Models, Animal; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D005260:Female; D006868:Hydrolysis; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects*; D008807:Mice, Inbred BALB C; D008826:Microbial Sensitivity Tests; D010457:Peptidoglycan / Q000187:drug effects; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D013203:Staphylococcal Infections / Q000188:drug therapy* / Q000382:microbiology; D016019:Survival Analysis; D016896:Treatment Outcome
chemical_list: D000900:Anti-Bacterial Agents; D010457:Peptidoglycan; D011994:Recombinant Proteins; D010450:Endopeptidases; C001143:endolysin
keywords: antibiotic resistance; antimicrobial; biofilm; peptidoglycan hydrolase
references: J Dairy Sci. 2000 Apr;83(4):855-62
references: 10791803
references: Appl Environ Microbiol. 2006 Apr;72(4):2988-96
references: 16598006
references: Antimicrob Agents Chemother. 2000 Oct;44(10):2845-7
references: 10991869
references: Nat Biotechnol. 2001 Jan;19(1):66-70
references: 11135555
references: Antimicrob Agents Chemother. 2001 Apr;45(4):999-1007
references: 11257008
references: Infect Immun. 2002 Jan;70(1):204-10
references: 11748184
references: Mol Microbiol. 2002 Mar;43(6):1367-78
references: 11952892
references: J Mammary Gland Biol Neoplasia. 2002 Apr;7(2):135-46
references: 12463736
references: Nat Med. 2004 Mar;10(3):243-5
references: 14758355
references: Microbiology. 2004 Jul;150(Pt 7):2079-87
references: 15256551
references: J Bacteriol. 2006 Sep;188(17):6286-97
references: 16923896
references: Appl Environ Microbiol. 2007 Jan;73(1):347-52
references: 17085695
references: JAMA. 2007 Oct 17;298(15):1763-71
references: 17940231
references: Int J Med Microbiol. 2008 Jul;298(5-6):505-13
references: 18221914
references: FEMS Microbiol Lett. 2008 Oct;287(1):22-33
references: 18673393
references: J Biotechnol. 2009 Jan 1;139(1):118-23
references: 18940209
references: Crit Care Med. 2009 Feb;37(2):642-9
references: 19114881
references: FEMS Microbiol Lett. 2009 May;294(1):52-60
references: 19493008
references: Appl Environ Microbiol. 2009 Jul;75(13):4483-90
references: 19411410
references: Gene. 2009 Aug 15;443(1-2):32-41
references: 19422893
references: Antimicrob Agents Chemother. 2010 Apr;54(4):1603-12
references: 20086153
references: Appl Microbiol Biotechnol. 2010 May;86(5):1439-49
references: 20013118
references: Int J Med Microbiol. 2010 Aug;300(6):357-62
references: 20452280
references: Virology. 2010 Nov 25;407(2):381-90
references: 20869739
references: Proc Natl Acad Sci U S A. 2010 Nov 2;107(44):18991-6
references: 20944066
references: Curr Opin Biotechnol. 2011 Apr;22(2):172-9
references: 21109420
references: Bioeng Bugs. 2010 Nov-Dec;1(6):404-7
references: 21468207
references: Antimicrob Agents Chemother. 2011 Sep;55(9):4416-9
references: 21746950
references: Mol Microbiol. 2011 Sep;81(6):1419-32
references: 21790805
references: Adv Virus Res. 2012;83:299-365
references: 22748813
references: Future Microbiol. 2012 Oct;7(10):1147-71
references: 23030422
references: J Biotechnol. 2012 Dec 31;162(2-3):289-98
references: 23026556
references: Appl Microbiol Biotechnol. 2013 Apr;97(8):3449-56
references: 22777279
references: Proc Natl Acad Sci U S A. 1964 Mar;51:414-21
references: 14171453
references: J Biol Chem. 1949 Nov;181(1):149-51
references: 15390401
references: Biochim Biophys Acta. 1965 Feb 15;97:242-50
references: 14292833
references: Antimicrob Agents Chemother. 1974 Aug;6(2):156-65
references: 15828186
references: Vet Microbiol. 2005 May 20;107(3-4):295-9
references: 15863290
references: Infect Immun. 2005 Jun;73(6):3502-11
references: 15908379
references: Gene. 2005 May 23;351:97-108
references: 15848115
references: Emerg Infect Dis. 2005 Jun;11(6):928-30
references: 15963289
references: Curr Opin Microbiol. 2005 Aug;8(4):480-7
references: 15979390
references: Antimicrob Agents Chemother. 2005 Aug;49(8):3256-63
references: 16048934
references: Science. 1994 Apr 15;264(5157):388-93
references: 8153626
references: FEMS Microbiol Lett. 1998 May 15;162(2):265-74
references: 9627962
references: Trends Biochem Sci. 1999 Apr;24(4):132-3
references: 10322416
references: Appl Environ Microbiol. 2000 Jul;66(7):2951-8
references: 10877791
references: J Clin Microbiol. 1985 Dec;22(6):996-1006
references: 3905855
references: J Bacteriol. 1990 Oct;172(10):5783-8
references: 1976618
references: Infect Immun. 2005 Oct;73(10):6868-76
references: 16177366
references: J Bacteriol. 2005 Oct;187(20):7161-4
references: 16199588
references: Antimicrob Agents Chemother. 2005 Nov;49(11):4789-92
references: 16251333
references: Clin Infect Dis. 2006 Feb 1;42(3):389-91
references: 16392087
references: Bacteriol Rev. 1972 Dec;36(4):407-77
references: 4568761
references: Antimicrob Agents Chemother. 1982 Apr;21(4):531-5
references: 7081976
references: J Bacteriol. 2006 Apr;188(7):2463-72
references: 16547033
references: Exp Biol Med (Maywood). 2006 Apr;231(4):366-77
references: 16565432
references: Protein Expr Purif. 2006 Apr;46(2):483-8
references: 16290005
references: J Biol Chem. 1999 May 28;274(22):15847-56
references: 10336488