This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Contreras, B. G. Articles by Vandamme, E. J. Search for Related Content PubMed PubMed Citation Articles by Contreras, B. G. Articles by Vandamme, E. J. Agricola Articles by Contreras, B. G. Articles by Vandamme, E. J.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., Jan 1997, 13-20, Vol 63, No. 1
Copyright © 1997, American Society for Microbiology

Isolation, purification, and amino acid sequence of lactobin A, one of the two bacteriocins produced by Lactobacillus amylovorus LMG P-13139

BG Contreras, L De Vuyst, B Devreese, K Busanyova, J Raymaeckers, F Bosman, E Sablon and EJ Vandamme
Laboratorium voor Industriele Microbiologie en Biokatalyse, Universiteit Gent, Belgium.

Lactobacillus amylovorus LMG P-13139, isolated from corn steep liquor, produces two bactericidal peptides with respective estimated molecular masses of 4.5 and 6.0 kDa upon denaturing sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The antimicrobial activity detected in the fermentation supernatant fraction of L. amylovorus LMG P-13139 was heat stable (20 min, 121 degrees C), displayed a narrow inhibitory spectrum, and was sensitive to proteinase K, trypsin, and alpha-chymotrypsin but insensitive to alpha-amylase, lysozyme, catalase, and lipase. The 4.5-kDa bacteriocin was purified and characterized and designated lactobin A. Lactobin A was isolated as a floating pellicle from culture supernatant brought to 35% saturation with ammonium sulfate. Upon this ammonium sulfate treatment, crude lactobin A was incorporated, together with Tween 80 as a major contaminant, in high-molecular-mass complexes sized at approximately 670 kDa by gel filtration chromatography. Contaminating fatty acids were removed from these micelles by a simple one-step methanol- chloroform extraction without loss of activity. Both inhibitory peptides were separated in an isocratic isopropanol gradient on a PepRPC 5/5 reversed-phase column, and both peptides retained activity towards Lactobacillus helveticus ATCC 15009 upon separation. Lactobin A has a molecular mass determined by electrospray mass spectrometry of 4,879 +/- 0.69 Da. Its peptide chain contains 50 unmodified amino acids, of which 26% are glycine residues and 40% are hydrophobic residues (A, V, L, I, and P). It displays the highest structural homology (42% identity and 28% similarity) with the lafX gene product, encoded by the second open reading frame of the lactacin F operon. These data strongly indicate that lactobin A belongs to the class IIb bacteriocins according to the classification of Klaenhammer.

This article has been cited by other articles:

• Line, J. E., Svetoch, E. A., Eruslanov, B. V., Perelygin, V. V., Mitsevich, E. V., Mitsevich, I. P., Levchuk, V. P., Svetoch, O. E., Seal, B. S., Siragusa, G. R., Stern, N. J. (2008). Isolation and Purification of Enterocin E-760 with Broad Antimicrobial Activity against Gram-Positive and Gram-Negative Bacteria. Antimicrob. Agents Chemother. 52: 1094-1100 [Abstract] [Full Text]  
• Zendo, T., Koga, S., Shigeri, Y., Nakayama, J., Sonomoto, K. (2006). Lactococcin Q, a Novel Two-Peptide Bacteriocin Produced by Lactococcus lactis QU 4.. Appl. Environ. Microbiol. 72: 3383-3389 [Abstract] [Full Text]  
• Kawai, Y., Ishii, Y., Arakawa, K., Uemura, K., Saitoh, B., Nishimura, J., Kitazawa, H., Yamazaki, Y., Tateno, Y., Itoh, T., Saito, T. (2004). Structural and Functional Differences in Two Cyclic Bacteriocins with the Same Sequences Produced by Lactobacilli. Appl. Environ. Microbiol. 70: 2906-2911 [Abstract] [Full Text]  
• Uteng, M., Hauge, H. H., Brondz, I., Nissen-Meyer, J., Fimland, G. (2002). Rapid Two-Step Procedure for Large-Scale Purification of Pediocin-Like Bacteriocins and Other Cationic Antimicrobial Peptides from Complex Culture Medium. Appl. Environ. Microbiol. 68: 952-956 [Abstract] [Full Text]  
• Cintas, L. M., Casaus, P., Herranz, C., Håvarstein, L. S., Holo, H., Hernández, P. E., Nes, I. F. (2000). Biochemical and Genetic Evidence that Enterococcus faecium L50 Produces Enterocins L50A and L50B, the sec-Dependent Enterocin P, and a Novel Bacteriocin Secreted without an N-Terminal Extension Termed Enterocin Q. J. Bacteriol. 182: 6806-6814 [Abstract] [Full Text]  
• Anderssen, E. L., Diep, D. B., Nes, I. F., Eijsink, V. G. H., Nissen-Meyer, J. (1998). Antagonistic Activity of Lactobacillus plantarum C11: Two New Two-Peptide Bacteriocins, Plantaricins EF and JK, and the Induction Factor Plantaricin A. Appl. Environ. Microbiol. 64: 2269-2272 [Abstract] [Full Text]