Purification and characterisation of acidocin D20079, a bacteriocin produced by Lactobacillus acidophilus DSM 20079.
Bacteriocins are natural antimicrobial agents produced by food fermentative bacteria. Lactobacillus acidophilus DSM 20079 produces a small bacteriocin, with a molecular mass of 6.6 kDa, designated acidocin D20079. This antimicrobial peptide was extremely heat-stable (30 min at 121 degrees C) and was active over a wide pH range. It was found to be sensitive to proteolytic enzymes (trypsin, ficin, pepsin, papain, and proteinase K). Acidocin D20079 has a narrow inhibitory spectrum restricted to the genus Lactobacillus which includes L. sakei NCDO 2714, an organism known to cause anaerobic spoilage of vacuum-packaged meat. Maximum production of acidocin D20079 in MRS broth was detected at pH 6.0, and the peptide was purified by ammonium sulphate precipitation followed by sequential cation exchange and hydrophobic interaction chromatography. Purified acidocin D20079 spontaneously formed spherulite crystals during dialysis. As the N-terminus was found to be blocked for sequencing, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was used to determine a partial sequence, and the molecular mass of the bacteriocin in the formed crystals (6.6 kDa). Estimates of the molecular weight of the partially purified peptide, using tricine-SDS-PAGE, in which bacteriocin activity was confirmed by overlayer techniques were in accordance with this value.
Deraz
Sahar F
SF
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, S-22100 Lund, Sweden.
Karlsson
Eva Nordberg
EN
Hedström
Martin
M
Andersson
Maria M
MM
Mattiasson
Bo
B
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007779:Lactobacillus acidophilus / Q000145:classification* / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013045:Species Specificity
D001430:Bacteriocins; C504214:acidocin D20079, Lactobacillus acidophilus