title: Bacteriophage phi11 lysin: Physicochemical characterization and comparison with phage phi80α lysin.
abstract: Phage lytic enzymes are promising antimicrobial agents. Lysins of phages phi11 (LysPhi11) and phi80α (LysPhi80α) can lyse (destroy) cells of antibiotic-resistant strains of Staphylococcus aureus. Stability of enzymes is one of the parameters making their practical use possible. The objectives of the study were to investigate the stability of lysins of phages phi11 and phi80α in storage and functioning conditions, to identify optimum storage conditions and causes of inactivation. Stability of the recombinant LysPhi11 and LysPhi80α was studied using turbidimetry. CD-spectroscopy, dynamic light scattering, and electrophoresis were used to identify causes of inactivation. At 37°C, pH 7.5 and concentration of NaCl not higher than 150mM, LysPhi11 molecules contain a high percentage of random coils (43%). However, in spite of this the enzyme has high activity (0.4-0.8OD600nms(-1)mg(-1)). In storage conditions (4°C and 22°C, pH 6.0-9.0, 10-500mM NaCl) LysPhi11 is inactivated by a monomolecular mechanism. The optimum storage conditions for LysPhi11 (4°C, pH 6.0-7.5, 10mM NaCl) were selected under which the time of the enzyme half-inactivation is 120-160 days. LysPhi80α stability is insufficient: at 37°C the enzyme loses half of its activity almost immediately; at 4°C and 22°C the time of half-inactivation of LysPhi80α varies in the range from several hours to 3 days. Despite the common properties in the manifestation of antistaphylococcal activity the kinetic behavior of the enzymes is different. LysPhi11 is a more promising candidate to be used as an antimicrobial agent.
authors.affiliation: Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia. Electronic address: luboff.filatova@gmail.com.
authors.forename: Lyubov Y
authors.initials: LY
authors.lastname: Filatova
authors.affiliation: Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, NEA, ARS, USDA, Beltsville, MD, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Donovan
authors.affiliation: Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, NEA, ARS, USDA, Beltsville, MD, USA.
authors.forename: Juli
authors.initials: J
authors.lastname: Foster-Frey
authors.affiliation: FBSE SRC of Virology & Bioengineering "Vector", Novosibirsk, Russia.
authors.forename: Vladimir G
authors.initials: VG
authors.lastname: Pugachev
authors.affiliation: I.M Sechenov First Moscow State Medical University, Moscow, Russia.
authors.forename: Natalia F
authors.initials: NF
authors.lastname: Dmitrieva
authors.affiliation: Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia.
authors.forename: Tatiana A
authors.initials: TA
authors.lastname: Chubar
authors.affiliation: Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia; Pirogov Russian National Research Medical University (RNRMU), Moscow, Russia.
authors.forename: Natalia L
authors.initials: NL
authors.lastname: Klyachko
authors.affiliation: Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia; Division of Molecular Pharmaceutics, Center for Nanotechnology in Drug Delivery, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, USA.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Kabanov
mesh_terms: D002118:Calcium / Q000378:metabolism; D004356:Drug Storage; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D009994:Osmolar Concentration; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000737:chemistry; D013045:Species Specificity; D013204:Staphylococcus Phages / Q000201:enzymology*; D013211:Staphylococcus aureus / Q000821:virology; D014764:Viral Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D012965:Sodium Chloride; D008274:Magnesium; D002118:Calcium
keywords: Activity; Bacteriolytic enzymes; Phages; Stability; Staphylococcal infections; Structure