Kinetics of inactivation of staphylolytic enzymes: Qualitative and quantitative description

Lyubov Filatova; David Donovan; Steven Swift; Vladimir Pugachev; Georgy Emelianov; Tatiana Chubar; Natalia Klaychko

doi:10.1016/j.biochi.2019.04.005

Kinetics of inactivation of staphylolytic enzymes: Qualitative and quantitative description

Biochimie. 2019 Jul:162:77-87. doi: 10.1016/j.biochi.2019.04.005. Epub 2019 Apr 6.

Authors

Lyubov Filatova¹, David Donovan², Steven Swift², Vladimir Pugachev³, Georgy Emelianov⁴, Tatiana Chubar⁴, Natalia Klaychko⁵

Affiliations

¹ Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia. Electronic address: luboff.filatova@gmail.com.
² Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, NEA, ARS, USDA, Beltsville, MD, USA.
³ Federal Budget Institution of Science, State Research Center of Virology & Bioengineering "Vector", Novosibirsk, Russia.
⁴ Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, Moscow, Russia.
⁵ 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.

PMID: 30965078
DOI: 10.1016/j.biochi.2019.04.005

Abstract

Lysin 2638aR and chimeric Ply187AN-KSH3b fusion protein are capable of lysing antibiotic-resistant strains of Staphylococcus aureus and are promising alternatives to antibiotics. Studies on the stability and structure of lysins 2638aR and Ply187AN-KSH3b are important for assessing the feasibility of their practical use. Both lysins are highly active at physiological pH (7.5) and at low salt content (the concentration of NaCl in the reaction medium is not more than 250 mM). Lysins are inactivated by a monomolecular mechanism and have high stability at 4 °C (storage temperature). The maximum value of the half-inactivation time for lysin 2638aR is 190-200 days (500-1000 mM NaCl, pH 6.0-7.5), for lysin Ply187AN-KSH3b is 320-340 days (10-1000 mM NaCl, pH 6.0). The lysins are pretty stable in human blood serum (the half-inactivation time is 0.5-2 h) at 37 °C. The lysins undergo denaturation in large part due to the destruction of the α-helices at temperatures above 40 °C.

Keywords: Bacteriophage endolysin; Chimeric lysin; Stability; Staphylococcus aureus; Structure.

MeSH terms

Cations / chemistry
Enzyme Stability
Hydrogen-Ion Concentration
Kinetics
N-Acetylmuramoyl-L-alanine Amidase / chemistry*
Recombinant Fusion Proteins / chemistry
Sodium Chloride / chemistry
Staphylococcus Phages / enzymology*
Temperature

Substances

Cations
Recombinant Fusion Proteins
Sodium Chloride
N-Acetylmuramoyl-L-alanine Amidase