Purification and Characterization of Enterocin LR/6, a Bacteriocin from Enterococcus faecium LR/6

• Published in: Applied biochemistry and biotechnology Vol. 160; no. 1; pp. 40 - 49
• Main Authors: Kumar, Manoj, Tiwari, Santosh Kumar, Srivastava, Sheela
• Format: Journal Article
• Language: English
• Published: New York New York : Humana Press Inc 2010; Humana Press Inc; Springer; Springer Nature B.V
• Subjects: Amino Acid Sequence; Ammonium; ammonium sulfate; Anti-Infective Agents - chemistry; Anti-Infective Agents - isolation & purification; Anti-Infective Agents - metabolism; Anti-Infective Agents - pharmacology; antibacterial properties; Antibiotics; Bacteria; Bacteriocins - chemistry; Bacteriocins - isolation & purification; Bacteriocins - metabolism; Bacteriocins - pharmacology; Biochemistry; Biological and medical sciences; Biotechnology; Cell Survival - drug effects; chemical precipitation; Chemistry; Chemistry and Materials Science; enterocins; Enterococcus faecium; Enterococcus faecium - chemistry; Enzymes; Filtrate; Fundamental and applied biological sciences. Psychology; high performance liquid chromatography; High temperature; Hot Temperature; Hydrogen-Ion Concentration; Hydrolysis; Ionization; Kinetics; Liquid chromatography; Low temperature; Mass spectrometry; Micrococcus luteus; Micrococcus luteus - cytology; Micrococcus luteus - drug effects; Mode of action; Molecular Weight; Organic Chemicals - pharmacology; Organic solvents; Peptide Hydrolases - metabolism; Protein Stability - drug effects; Proteins; purification; Solvents - pharmacology; storage temperature; Sulfates; Surface-Active Agents - pharmacology; Enterocin LR/6; Characterization; Purification; Mode of action; Streptococcaceae; Enterococcus faecium; Bacteriocin; Bacteria; Micrococcales; Mechanism of action; Entenococcus faecium
• ISSN: 0273-2289; 1559-0291
• DOI: 10.1007/s12010-009-8586-z

Enterocin LR/6, a bacteriocin obtained from the culture filtrate of Enterococcus faecium strain LR/6, has been purified to homogeneity using ammonium sulfate precipitation, cation-exchange chromatography, gel-filtration, and checked on reverse-phase high-performance liquid chromatography. It is active at high temperatures (boiling as well as autoclaving) and over a wide range of pH (2.0-8.0). Also, it is sensitive to a number of proteolytic enzymes but is stable in the presence of surfactants and organic solvents. The protein could be stored at least up to 1 year at low temperatures (4 °C and −20 °C) without any loss of activity. The N-terminal sequence of enterocin LR/6 showed no homology with known enterocins or other bacteriocins present in the database, suggesting it to be a novel enterocin. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed its mass to be ∼6.1 kDa. It showed a bactericidal mode of action against indicator strain, Micrococcus luteus.