Bazı Laktik Asit Bakterilerinin Fizyolojik, Biyokimyasal, Plazmit DNA ve Protein Profil Özelliklerinin İncelenmesi

Bu çalısmada, toplam 36 adet laktik asit bakterisi kefir, beyaz peynir, kasar peyniri ve sucuktan izole edilmistir. Izole edilen kültürler biyokimyasal testler ve API 50 CH kiti ile tanımlanmıs ve toplam protein profilleri Sodyum Dodesil Sülfat Poliakrilamid Jel Elektroforezi (SDS-PAGE) ile belirlenmistir. Tanımlama testleri sonucunda 36 adet laktik asit bakterisi, Leuconostoc cremoris (1 sus), Leu. mesenteroides (1 sus), Lactobacillus brevis (1 sus), Lb. casei (2 sus), Lb. lactis (2 sus), Lb. plantarum (2 sus), Lb. helveticus (3 sus), Lactococcus cremoris (3 sus), Lac. lactis (3 sus), Streptococcus durans (1 sus), Strep. thermophilus (2 sus), Pediococcus acidilactici (2 sus), P. pentosaceus (4 sus) ve P. dextrinicus (9 sus) olarak tanımlanmıstır. 36 adet laktik asit bakteri susunun plazmit DNA incelemesinde, 21 adet susta 1-5 arasında degisen sayılarda plazmit DNA’ya rastlanılmıstır. Bu plazmit DNA’ların moleküler agırlıklarının 3.11-28.07 kb arasında oldugu belirlenmistir. 15 adet laktik asit bakterisinin ise plazmit DNA içermedikleri gözlenmistir.

Studying of Physiological, Biochemical, Plasmid DNA and Protein Profiles of Some Lactic Acid Bacteria (Turkish with English Abstract)

In this study, 36 lactic acid bacteria were isolated from kefir, White Pickled cheese, Kashar cheese and sucuk (a Turkish style fermented sausage). The isolated cultures were identified by biochemical tests and API 50 CH test kit. Total proteins of strains were carried on Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). 36 strains were identified as Leuconostoc cremoris(1 strain), Leu. mesenteroides(1), Lactobacillus brevis(1), Lb. casei(2), Lb. lactis(2), Lb. plantarum(2), Lb. helveticus(3), Lactococcuscremoris(3), Lac. lactis(3), Streptococcus durans(1), Strep. thermophilus(2), Pediococcus acidilactici(2), P.pentosaceus(4) and P. dextrinicus(9). Plasmid DNA profiles of lactic acid bacteria were examined and theirmolecular weights were identified. Results indicated that 21 strains carried 1–5 plasmids and their molecularweights ranged between 3.11 and 28.07 kb. Remaining 15 strains did not carry any plasmid DNA.

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  • 1. Hofvendahl K, Hahn-Hägerdal B. 2000 Factors affecting the fermentative lactic acid production from renewable resources. Enzyme Microb Technol, 26: 87–107.
  • 2. Tunail N, Köşker. 1989. Süt Mikrobiyolojisi. Ankara Üniversitesi Zıraat Fakültesi Yayınları No:1116 Ankara, 18–19.
  • 3. Zhu WM, Liu M, Wu DQ. 2000. Isolation and characterization of a new bacteriocin from Lactobacillus gasseri KT7. J Appl Microbiol, 88: 877–886.
  • 4. Woodford N (ed), Johnson AP. 1998. Plasmid Analysis. Humana Press, New Jersey, USA, 356 p.
  • 5. Turner PC, McLennon AG, Bates AD, White MRH. 2000. Instant Notes Molecular Biology. School of Biological Sciences, University of Liverpool, 103.
  • 6 Martinez-Bueno M, Valdivia E, Galvez A, Maqueda M. 2000. pS86 a new theta-replicating plasmid from Enterococcus faecalis. Curr Microbiol, 41: 257–261.
  • 7. Axelsson LT. 1993. Lactic acid bacteria: classification and physiology. In: Salminen S., von Wright, A., editors. Lactic acid bacteria. New York: Marcel Dekker, Inc., 1-72.
  • 8. Qhobela M, Leach JE, Claflin LE, Pearson DL.1991. Characterisation of starins of Xanthomonas campestris pv. holcicola by PAGE of membrane proteins and by REA and RFLP analysis of genomic DNA. Plant Disease, 75: 32–36.
  • 9. Harrigan W, McCance M. 1990. Laboratory Methods in Food and Dairy Microbiology (8th ed). London, UK: Academic Pres.
  • 10. Wood BJB, Holzapfel WH. 1995. The Genera of Lactic Acid Bacteria, vol. 2, Blackie Academic and Professional, Glasgow.
  • 11. Tan ZY, Xu XD, Wang ET, Gao TL, Romero EM, Chen WX. 1997. Phylogenetic and genetic relationships of Mesorhizobium tianshanense and related Rhizobia. Int J Syst Bacteriol, 47: 874–879.
  • 12. Laemmli UK. 1970. Cleavage of structural proteins during the assambly of the head of bacteriophage T4. Nature, 227: 680–685.
  • 13. Wolff K, Peters-van Rijn J. 1993. Rapid detection of genetic variability in chrysanthemum (Dendranthema grandiflora Tzvelev) using random primers. Heredity, 71: 335–341.
  • 14. O’sullivian DJ, Klaenhammer TR. 1993. Rapid miniprep isolation of high-quality plasmid DNA from Lactococcus and Lactobacillus spp. Appl Environ Microbiol 59: 2730–2733.
  • 15. Macrina FL, Tobian JA, Jones KR, Evans RP, Clewell DB. A cloning vector able to replicate in Escherichia coli and Streptococcus sangius. Gene, 19: 345–353.
  • 16. Sambrook J (ed), Fidsch EF, Maniatis T. 1989. Molecular Cloning a Laboratory Manual. Cold Spring Harbor Laboratory Pres, UK, 643p.
  • 17. Piodux M, Marshall VM, Zanoni P, Brooker B. 1990. Lactobacilli isolated from sugary kefir grains capable of polysaccharide production and minicell formation. J Appl Bacteriol 69: 311–320.
  • 18. Yuksekdag ZN, Beyatli Y, Aslim B. 2004. Determination of some characteristics coccoid forms of lactic acid bacteria isolated from Turkish kefirs with natural probiotic. Lebensm-Wiss Technol, 37: 663–667.
  • 19. Angulo L, Lopez E, Lema C. 1993. Micro flora present in kefir grain of the galician region (northwest of Spain). J Dairy Research, 60: 263–267.
  • 20. Marshall VM, Colc WM, Farrow, JA. 1984. A note on the heterofermentative Lactobacillus isolated from kefir grains. J Appl Bacteriol, 56: 503–505.
  • 21. Simova ED, Beskova DM, Angelov A, Hristozova TS, Frengova G, Sposov Z. Lactic acid bacteria and yeasts in kefir grains and kefir made from them. J Industrial Microbiol Biotechnol, 28: 1–6.
  • 22. Beskova DM, Simova ED, Simov ZI, Frengova GI, Spasov ZN. 2002. Pure cultures for making kefir. Food Microbiol, 19: 537–544.
  • 23. Sagdic O, Simsek B, Kucukoner E. 2003. Microbiological and physiochemical characteristics of Van herby cheese, a traditional Turkish dairy product. Milchwissenschaft, 58: 382–385.
  • 24. Papamanoli E, Tzanetakis N, Litopoulou-Tzanetaki E, Katzekidou P. 2003. Characterization of lactic acid bacteria isolated from a Grek dry-fermented sausage in respect of their technological and probiotic properties. Meat Sci, 65: 859–867.
  • 25. Flint SH, Brooks JD, Bremer PJ. 1997. The influence of cell surface properties of thermophilic streptococci on attachment to stainless steel. J Appl Microbiol 83: 508–517.
  • 26. Ercolini D, Moschetti G, Blaitta G, Coppola S. 2001. Behavior of variable V3 region from 16S rDNA of lactic acid bacteria in denaturing gradient gel electrophoresis. Curr Microbiol, 42: 199–202.
  • 27. Ozkaya FD, Xanthopoulos V, Tunail N, Tzanetaki EL.2001. Technologically important properties of lactic acid bacteria isolated from Beyaz cheese made from raw ewes’ milk. J Appl Microbiol 91: 861–870.
  • 28. Andrighetto C, Knijff E, Lombardi A, Torriani S, Vancanneyt M, Kersters K, Swings J, Dellaglio F.2001. Phenotypic and genetic diversity of enterococci isolated from Italian cheeses. J Dairy Res, 68: 303–316.
  • 29. Sánchez I, Seseňa S, Palop L. 2003. Identification lactic acid bacteria from spontaneous fermentation of Almagro eggplants SDS-PAGE whole cell protein fingerprinting. Int J Food Microbiol. 82: 181–189.
  • 30. Leisner JJ, Pot B, Christensen H, Rusul G, Olsen JE, Wee BW, Muhammad K, Ghali HM. 1999. Identification of lactic acid bacteria from ohili bo, a Malaysian food ingredient. Appl Environ Microbiol, 65 (2): 599–605.
  • 31. Hertel C, Lutwik W, Pot B, Kersters K, Schleıfer KH. 1993. Differentiation of lactobacilli occurring in fermented milk products by using oligonucleotide probes and electrophoretic protein profiles. System Appl Microbiol 16: 463–467.
  • 32. De Angelis M, Corsetti A, Tosti N, Rossi J, Corbo MR, Gobbetti M. Characterization of non-starter lactic acid bacteria from Italian ewe cheeses based on phenotypic, genotypic and cell wall protein analyses. Appl Environ Microbiol, 67: 2011–2020.
  • 33. Arda M. 2000. Temel Mikrobiyoloji. İkinci baskı. Medisan Yayınları, Ankara, 490–548.
  • 34. Nakamura S, Tagawa Y, Miyamoto T, Kataoka K. 1992. Plasmid profiles of lactose-, citrate- and proteinase-negative mutants of Lactococcus lactis subsp. lactis biovar diacetylactis strain N-7. Milchwissenschaft, 47: 358–361.
  • 35. McKay LL. 1983. Functional properties of plasmids in lactic streptococci. Anton. Van Leeuw, 49: 259–274.
  • 36. van Belkum MJ, Hayema B, Geis A, Kok J, Venema G. 1989. Cloning of two bacteriocin genes from a lactococcal bacreiocin plasmid. Appl Environ Microbiol, 55: 1187–1191.
  • 37. Hastings JW, Stiles ME. 1991. Antibiosis of Leuconostoc gelidum isolated from meat. J Appl Bacteriol 70: 127–134.
  • 38. Kim WJ, Ray B, Johnson MC. 1992. Plasmid transfers by conjuction and electroporation in Pediococcus acidilactici. J Appl Bacteriol, 72: 201–207.
  • 39. Leblanc DJ, Crow VL, Lee LN. 1980. In: Plasmid and transposon environmental effects and maintenaces mecahanisms. Stutard W (chief ed), Academic Press, New York, USA, pp. 31.