Characterization, production, and purification of leucocin H, a two-peptide bacteriocin from Leuconostoc MF215B.
Leuconostoc MF215B was found to produce a two-peptide bacteriocin referred to as leucocin H. The two peptides were termed leucocin Halpha and leucocin Hbeta. When acting together, they inhibit, among others, Listeria monocytogenes, Bacillus cereus, and Clostridium perfringens. Production of leucocin H in growth medium takes place at temperatures down to 6 degrees C and at pH below 7. The highest activity of leucocin H in growth medium was demonstrated in the late exponential growth phase. The bacteriocin was purified by precipitation with ammonium sulfate, ion-exchange (SP Sepharose) and reverse phase chromatography. Upon purification, specific activity increased 10(5)-fold, and the final specific activity was 2 x 10(7) BU/OD280. Amino acid composition analyses of leucocin Halpha and leucocin Hbeta indicated that both peptides consisted of around 40 amino acid residues. Their N-termini were blocked for Edman degradation, and the methionin residues of leucocin Hbeta did not respond to Cyanogen Bromide (CNBr) cleavage. Absorbance at 280 nm indicated the presence of tryptophan residues and tryptophan-fracturing opened for partial sequencing by Edman degradation. From leucocin Halpha, the sequence of 20 amino acids was obtained; from leucocin Hbeta the sequence of 28 amino acid residues was obtained. No sequence homology to other known bacteriocins could be demonstrated. It also appeared that the two peptides themselves shared little or no sequence homology. The presence of soy oil did not affect the activity of leucocin H in agar.
Blom
H
H
MATFORSK, Norwegian Food Research Institute, Oslovn. 1, N-1430 As, Norway.
Katla
T
T
Holck
A
A
Sletten
K
K
Axelsson
L
L
Holo
H
H
D000595:Amino Acid Sequence; D001409:Bacillus cereus / Q000187:drug effects; D001430:Bacteriocins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D003016:Clostridium perfringens / Q000187:drug effects; D007934:Leuconostoc / Q000378:metabolism*; D008089:Listeria monocytogenes / Q000187:drug effects; D008969:Molecular Sequence Data
D001430:Bacteriocins; C120438:leucocin H