Didem ŞAHİNGİL, Hilal İŞLEROĞLU, Zeliha YILDIRIM, Mustafa AKÇELİK, Metin YILDIRIM

Characterization of lactococcin BZ produced by Lactococcus lactis subsp. lactis BZ isolated from boza

A bacterium isolated from boza produced in Turkey was identified, and the physico-chemical and microbiological properties of its inhibitory compound were characterized. The isolate was identified as Lactococcus lactis subsp. lactis, based on morphology, physiology, carbohydrate fermentation, the fatty acid profile, and 16S rDNA gene sequence homology. The antimicrobial compound produced by the microorganism, lactococcin BZ, was sensitive to papain, pepsin, trypsin, and b-mercaptoethanol, but was resistant to catalase, amylase, lipase, organic solvents (methanol, chloroform, etc.), detergents (SDS, urea, Tween-80, Triton X-100), and EDTA. Lactococcin BZ was active against several gram-positive and gram-negative foodborne pathogens and food spoilage bacteria. Lactococcin BZ maintained its activity after high-heat treatment (90 °C for 30 min), at acidic and neutral pHs (2.0-7.0), and after storage at -20 to -80 °C for 3 months in lyophilized form. Lactococcin BZ was produced at the maximum level in MRS broth with an inoculum volume of 0.1%, an initial pH of 7.0, and an incubation temperature of 25 °C. Bacteriocin production began during the logarithmic phase and reached the maximum level during the early stationary phase. Its mode of action against Listeria monocytogenes was bactericidal and its molecular weight was about 5500 Da, as determined using tricine SDS-PAGE. Lactococcus lactis subsp. lactis BZ or its bacteriocin, which has a wide inhibitory spectrum, has the potential for use as a biopreservative in food products.

Anahtar Kelimeler:

Key words: Bacteriocin, boza, Lactococcus lactis subsp. lactis, lactococcin BZ, antimicrobial compound

Characterization of lactococcin BZ produced by Lactococcus lactis subsp. lactis BZ isolated from boza

Keywords:

Key words: Bacteriocin, boza, Lactococcus lactis subsp. lactis, lactococcin BZ, antimicrobial compound,

PDF

___

Magnusson J, Schnürer J. Lactobacillus coryniformis subsp. coryniformis proteinaceous antifungal compound. Appl Environ Microbiol 67: 1-5, 2001.
De Vuyst L, Leroy F. Bacteriocins from lactic acid bacteria: production, purification, and food applications. J Mol Microbiol Biotechnol 13: 194-199, 2007.
Nes I, Johnsborg O. Exploration of antimicrobial potential in LAB by genomics. Curr Opin Biotechnol 15: 1-5, 2004.
Cotter PD, Hill C, Ross RP. Bacteriocins: developing innate immunity for food. Nat Rev Microbiol 3: 777-788, 2005.
Ivanova I, Kabadjova P, Pantev A et al. Detection, purification and partial characterization of a novel bacteriocin substance produced by Lactococcus lactis subsp. lactis B14 isolated from Boza Bulgarian traditional cereal beverage. Bio Fundamen Appl 41: 47-53, 2000.
Todorov SD, Dicks LMT. Screening for bacteriocin-producing lactic acid bacteria from Boza, a traditional cereal beverage from Bulgaria. Process Biochem 41: 11-19, 2006.
Todorov SD, Danova ST, Van Reenen CA et al. Characterization of bacteriocin HV219, produced by Lactococcus lactis subsp. lactis HV219 isolated from human vaginal secretions. J Basic Microbiol 46: 226-238, 2006.
Cleveland J, Montville TJ, Nes IF et al. Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 71: 1- 20, 2001.
Chen H, Hoover DG. Bacteriocins and their food applications. Compr Rev Food Sci Food Safety 2: 82-100, 2003.
Deegan LH, Cotter PD, Hill C et al. Bacteriocins: biological tools for bio-preservation and shelf-life extension. Int Dairy J 16: 1058-1071, 2006.
Gálvez A, Abriouel H, López RL et al. Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol 120: 51-70, 2007.
Hurst A. Nisin. Adv Appl Microbiol 27: 85-123, 1981.
Piard JC, Muriana PM, Desmazeaud MJ et al. Purification and partial characterization of lacticin 481, a lanthionine-containing bacteriocin produced by Lactococcus lactis subsp. lactis CNRZ 481. Appl Environ Microbiol 58: 279-284, 1992.
Ryan MP, Rea MC, Hill C et al. An application in cheddar cheese manufacture for a strain of Lactococcus lactis producing a novel broad-spectrum bacteriocin, lacticin 3147. Appl Environ Microbiol 62: 612-619, 1996.
Yildirim Z, Johnson MG. Detection and characterization of a bacteriocin produced by Lactococcus lactis subsp. cremoris R isolated from radish. Lett Appl Microbiol 26: 297-304, 1998.
Choi HJ, Cheigh CI, Kim SB et al. Production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi. J Appl Microbiol 88: 563-571, 2000.
Ferchichi M, Frere J, Mabrouk K et al. Lactocin MMFII, a novel class IIa bacteriocin produced by Lactococcus lactis MMFII, isolated from Tunisian dairy product. FEMS Microbiol Lett 205: 49-55, 2001.
Oscariz JC, Pisabarro AG. Classification and mode of action of membrane-active bacteriocins produced by Gram positive bacteria. Int Microbiol 4: 13-19, 2001.
Garneau S, Martin NI, Vederas JC. Two-peptide bacteriocins produced by lactic acid bacteria. Biochimie 84: 577-592, 2002.
Onda T, Yanagida F, Tsuji M et al. Production and purification of a bacteriocin peptide produced by Lactococcus spp. strain GM005, isolated from Miso-Paste. Int J Food Microbiol 87: 153- 159, 2003.
Millette M, Dupont C, Archambault D et al. Partial characterization of bacteriocins produced by human Lactococcus lactis and Pediococccus acidilactici isolates. J Appl Microbiol 102: 274-282, 2007.
Tükel Ç, Avsaroglu MD, Şimşek Ö et al. Isolation and partial characterization of a novel bacteriocin produced by Lactococcus lactis ssp. lactis MC38. J Food Safety 27: 17-29, 2007.
Kabadjova P, Gotcheva I, Ivanova I et al. Investigation of bacteriocin activity of lactic acid bacteria isolated from Boza. Biotechnol Biotechnol Eq 14: 56-59, 2000.
Todorov SD, Dicks LMT. Characterization of mesentericin ST99, a bacteriocin produced by Leuconostoc mesenteroides subsp. dextranicum ST99 isolated from Boza. J Ind Microbiol Biotechnol 31: 323-329, 2004.
Todorov SD, Dicks LMT, Effect of growth medium on bacteriocin production by Lactobacillus plantarum ST194BZ, a strain isolated from Boza. Food Technol Biotechnol 43: 165- 173, 2005.
Todorov SD, Dicks LMT. Pediocin ST18, an anti-listerial bacteriocin produced by Pediococcus pentosaceus ST18 isolated from Boza, a traditional cereal beverage from Bulgaria. Process Biochem 40: 365-370, 2005.
Todorov SD, Nyati H, Meincken M et al. Partial characterization of bacteriocin AMA-K, produced by Lactobacillus plantarum AMA-K isolated from naturally fermented milk from Zimbabwe. Food Control 18: 656-664, 2007.
von Mollendorff JW, Todorov SD, Dicks LMT. Comparison of bacteriocins produced by lactic-acid bacteria isolated from Boza, a cereal-based fermented beverage from the Balkan Peninsula. Curr Microbiol 53: 209-216, 2006.
Mayr-Harting A, Hedges AJ, Berkley RW. Methods for studying bacteriocins. In: Noris JR, Ribbons NW eds. Methods in Microbiology. Academic Press; 1972: pp. 315-442.
Beasley SS, Saris PEJ. Nisin-producing Lactococcus lactis strains from human milk. Appl Environ Microbiol 70: 5051-5053, 2004.
Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain- terminating inhibitors. Proc Natl Acad Sci USA 74: 5463-5467, 1977.
Endo A, Okada S. Monitoring the lactic acid bacterial diversity during Shochu fermentation by PCR-denaturing gradient gel electrophoresis. J Biosci Bioeng 3: 216-221, 2005.
Bhunia A, Johnson MC, Ray B. Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici. J Appl Bacteriol 65: 261-268, 1988.
De Kwaadsteniet M, Todorov SD, Knoetze H et al. Characterization of a 3944 Dalton bacteriocin, produced by Enterococcus mundtii ST15, with activity against Gram-positive and Gram-negative bacteria. Int J Food Microbiol 105: 433-444, 2005.
Moreno FMR, Leisner JJ, Tee LK et al. Microbial analysis of Malaysian tempeh, and characterization of two bacteriocins produced by isolates of Enterococcus faecium. J Appl Microbiol 92: 147-157, 2002.
Bhunia AK, Johnson MG. A modified method to directly detect in SDS-PAGE the bacteriocin of Pediococcus acidilactici. Lett Appl Microbiol 15: 5-7, 1992.
Schagger H, Von Jagov G. Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368-379, 1987.
Mundt JO. Lactic acid streptococci. In: Sneath PHA et al. eds. Bergey’s Manual of Systematic Bacteriology. Vol 2, Baltimore: The Williams & Wilkins Co; 1986: pp. 1065-1066.
Schleifer KH, Kilpper-Balz R. Molecular and chemotaxonomic approaches to the classification of streptococci, enterococci and lactococci: a review. Syst Appl Microbiol 10: 1-19, 1987.
Samyn B, Martinez-Bueno M, Devreese B et al. The cyclic structure of the enterococcal peptide antibiotic AS-48. FEBS Lett 352: 87-90, 1994.
Ivanova I, Miteva V, Stefanova T et al. Characterization of a bacteriocin produced by Streptococcus thermophilus 81. Int J Food Microbiol 42: 147-158, 1998.
Ko SH, Ahn C. Bacteriocin production by Lactococcus lactis KCA2386 isolated from white kimachi. Food Sci Biotechnol 9: 263-269, 2000.
Lee NK, Paik HD. Partial characterization of lacticin NK24, a newly identified bacteriocin of Lactococcus lactis NK24 isolated from Jeot-Gal. Food Microbiol 18: 17-24, 2001.
Messi P, Bondi M, Sabia C et al. Detection and preliminary characterization of a bacteriocin (plantaricin 35d) produced by a Lactobacillus plantarum strain. Int J Food Microbiol 64: 193- 198, 2001.
Klaenhammer TR. Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol 12: 39-86, 1993.
Ahn J, Lee H, Joo Y et al. Purification and characterization of a bacteriocin produced by Lactococcus lactis subsp. lactis H-559 isolated from Kimchi. J Biosci Bioeng 88: 153-159, 1999.
Villani F, Aponte M, Blaiotta G et al. Detection and characterization of a bacteriocin, garviecin L1-5, produced by Lactococcus garvieae isolated from raw cow’s milk. J Appl Microbiol 90: 430-439, 2001.
Ponce AG, Moreira MR, del Vale CE et al. Preliminary characterization of bacteriocin-like substances from lactic acid bacteria isolated from organic leafy vegetables. LWT- Food Sci Technol 41: 432-441, 2008.
Kemperman R, Kuipers A, Karsens H et al. Identification and characterization of two novel clostridial bacteriocins, circularin a and closticin 574. J Appl Environ Microbiol 69: 1589-1597, 2003.
Drider D, Fimland G, Hechard Y et al. The continuing story of class IIa bacteriocins. Microbiol Mol Biol Rev 70: 564-582, 2006.
Yang R, Johnson MC, Ray B. Novel method to extract large amounts of bacteriocins from lactic acid bacteria. Appl Environ Microbiol 58: 3355-3359, 1992.
Ghrairi T, Frere J, Berjeaud JM et al. Lactococcin MMT24, a novel two-peptide bacteriocin produced by Lactococcus lactis isolated from rigouta cheese. Int J Food Microbiol 105: 389-398, 2005.
De Vuyst L, Callewaert R, Crabbe K. Primary metabolite kinetics of bacteriocins biosynthesis by Lactobacillus amylovorus and evidence for stimulation of bacteriocins production under unfavourable growth conditions. Microbiol 142: 817-827, 1996.
Aasen IM, Moreto T, Katla T et al. Influence of complex nutrients, temperature and pH on bacteriocins production by Lactobacillus sakei CCUG 42687. Appl Microbiol Biotechnol 53: 159-166, 2000.
Cheigh CI, Choi Kim SB, Pyun YR. Production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from Kimchi. J Appl Microbiol 88: 563-571, 2000.
Cintas LM, Casaus P, Herraz C et al. 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, 2000.
Svetoslav D, Todorov SD, Danova ST et al. Characterization of bacteriocin HV219, produced by Lactococcus lactis subsp. lactis HV219 isolated from human vaginal secretions. J Basic Microbiol 46: 226-238, 2006.
Moreno I, Lerayer ALS, Baldini VLS et al. Characterization of bacteriocins produced by Lactococcus lactis strains. Brazilian J Microbiol 31: 183-191, 2000.
Park SH, Itoh K, Kikuchi ENH et al. Identification and characteristics of nisin Z-producing Lactococcus lactis subsp. lactis isolated from Kimchi. Curr Microbiol 46: 385-388, 2003.