Praveen Pal Balgir, Puja Bhatia, Pushpinder Kaur and Vipin Kumar*
Department of Biotechnology, Punjabi University, Patiala-147002, Punjab, India.
*Directorate of Research, SVPUA&T, Meerut, 250110 (UP), INDIA
E-mail of corresponding author: puja17bt@gmail.com
Abstract: The DNA-intercalating agents, ethidium bromide and ascorbic acid (vitamin C), were used to eliminate plasmid DNA from Pediococcus acidilactici MTCC 5101, a lactic acid bacteria. The strain was grown in the presence of 0.03 mM ethidium bromide or 1 mM ascorbate for 18 hours at 42°C, which resulted in the loss of its ability to produce pediocin, a plasmid-specified trait. Agarose gel electrophoresis of plasmid DNA and spot-on-lawn assay showed concomitant loss of an 8.9 kb plasmid pCP289 coding for pediocin from these colonies. Since ascorbic acid is a readily available, shows less effect on cell viability, non-hazardous and more efficient compound in contrast to ethidium bromide, the possibility of its use in determining plasmid-encoded traits in food grade lactic acid bacteria is also proposed.
Key words: Curing, ethidium bromide, ascorbic acid, Pediococcus acidilactici MTCC 5101
REFERENCES
Amabile-Cuevas, C.F.; Pina-Zentella, R. and Wah-Laborde, M.E. (1991). Decreased resistance to antibiotics and plasmid loss in plasmid-carrying strains of Staphylococcus aureus treated with ascorbic acid. Mutat Res, 264: 119-25.
Anderson, D.G. and McKay, L.L. (1983). Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol 46: 549-52.
Beg, A.Z. and Ahmad, I. (2000). Effect of Plumbago zeylanica extract and certain curing agents on multidrug resistant bacteria of clinical origin. World Journal of Microbiology and Biotechno-logy 16: 841-844.
Bhunia, A.K.; Johnson, M.C. and Ray, B. (1988). Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici. J Appl Bacteriol, 65: 261-8
Bhunia, A.K.; Johnson, M.C.; Ray, B. and Belden, E.L. (1990). Antigenic property of pediocin AcH produced by Pediococcus acidilactici H. J Appl Bacteriol, 69: 211-5.
BOUANCHAUD,D.H.;SCAVIZZI, M.R. And CHABBERT, Y.A. (1968). Elimination by ethidium bromide of antibiotic abbert. resistance in enterobacter and staphylococci. J. Gen. Microbiol, 54: 417-425.
Cho, H.Y.; Yousef, A.E. and Yang, S.T. (1996) Continuous production of pediocin by immobilized Pediococcus acidilactici PO2 in a packed-bed bioreactor. Applied Microbiology and Biotechnology, 45: 589-594.
Coventry, M.J.; Muirhead, K. and Hickey, M.W. (1995). Partial characterisation of pediocin PO2 and comparison with nisin for biopreservation of meat products. Int J Food Microbiol, 26: 133-45.
Crameri, R., Davies, J. E., Hutter, R. (1986) Plasmid curing and generation of mutations induced with ethidium bromide in streptomycetes. J Gen Microbiol 132: 819-24.
Daeschel, M.A. and Klaenhammer, T.R. (1985). Association of a 13.6-Megadalton Plasmid in Pediococcus pentosaceus with Bacteriocin Activity. Appl. Environ. Microbiol, 50: 1538-1541
De Man, J.C.; Rogosa, M. andSharpe, M. E. (1960). A medium for the cultivation of Lactobacilli.
J. Appl. Bacl., 23: 130–135.
Gonzalez, C.F. and Kunka, B. S. (1987). Plasmid-Associated Bacteriocin Production and Sucrose Fermentation in Pediococcus acidilactici. Appl. Environ. Microbiol, 53: 2534-2538.
Gupta, R.K. and Batish, V.K. (1992). Protoplast-induced curing of bacteriocin plasmid in Lactococcus lactis subsp. lactis 484. Journal of Applied Microbiology 73: 337-341.
Halami, P.M.; Chandrashekar, A. and Joseph, R. (1999). Characterization of bacteriocinogenic strains of lactic acid bacteria in fowl and fish intestines and mushroom. Food Biotechnology 13: 121-136.
Halliwell,B. and Aruoma, O. I. (1991). DNA damageby oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS Lett 281: 9-19.
Imlay, J.A. and Linn, S. (1988). DNA damage and oxygen radical toxicity. Science 240: 1302-9 Jager, K., Harlander, S. (1992) Characterization of a bacteriocin from Pediococcus acidilactici PC and comparison of bacteriocin-producing strains using molecular typing procedures. Applied microbiology and biotechnology, 37: 631-637.
Jager, K. and Harlander, S. (1992). Characterization of a bacteriocin from Pediococcus acidilactici PC and comparison of bacteriocin-producing strains using molecular typing procedures. Applied microbiology and biotechnology, 37: 631-637.
Jamal, H.; Chua, K.H.; Frederick, D.; Mahmood Ameen, A. and Salmah, I. (2005). Plasmid DNA Analysis of Pasteurella Multocida Serotype B Isolated from Haemorrhagic Septicaemia Outbreaks in Malaysia. Malaysian Journal of Microbiology, 1: 35-39.
Kaur, B. and Balgir, P.P. (2007). Pediocin CP2 Gene Localisation to plasmid pCP289 of Pediococcus acidilactici MTCC 5101. Internet J. Microbiol. 3
Kim, C.H.; Ji, G.E. and Ahn, C. (2000). Purification and molecular characterization of a bacteriocin from Pediococcus sp. KCA 1303-10 isolated from fermented flat fish. Food Sci. Biotechnol, 9: 270-276.
Marugg, J.D.; Gonzalez, C.F.; Kunka, B.S.; Ledeboer, A.M.; Pucci, M.J.; Toonen, M.Y.; Walker, S.A.; Zoetmulder, L.C. and Vandenbergh,P.A.(1992).Cloning,expression, and nucleotide sequence of genes involved in production of pediocin PA-1, a bacteriocin from Pediococcus acidilactici PAC1.0. Appl. Environ. Microbiol, 58: 2360-7.
Miller, K. W., Ray, P., Steinmetz, T., Hanekamp, T., Ray, B. (2005). Gene organization and sequences of pediocin AcH/PA-1 production operons in Pediococcus and Lactobacillus plasmids. Lett. Appl. Microbiol, 40: 56-62.
Morgan, A.R.; Cone, R.L. and Elgert, T.M. (1976). The mechanism of DNA strand breakage by vitamin C and superoxide and the protective roles of catalase and superoxide dismutase. Nucleic Acids Res., 3: 1139-49.
Motlagh, A.; Bukhtiyarova, M. and Ray, B. (1994). Complete nucleotide sequence of pSMB 74, a plasmid encoding the production of pediocin AcH in Pediococcus acidilactici. Letters in applied microbiology, 18: 305-312.
Motlagh, A.M.; Bhunia, A.K.; Szostek, F.; Hansen, T.R.; Johnson, M.C. and Ray, B. (1992). Nucleotide and amino acid sequence of pap-gene (pediocin AcH production) in Pediococcus acidilactici H. Lett Appl Microbiol, 15: 45-8.
Osmanagaoglu, O.; Beyatli, Y.; Gunduz, U. and Sacilik, S.C. (2000). Analysis of the genetic determinant for production of the pediocin P of Pediococcus pentosaceus Pep1. J. Basic Microbiol, 40: 233-41.
Piva, A. and Headon, D.R. (1994). Pediocin A, a bacteriocinproduced by Pediococcus pentosaceus FBB61. Microbiology 140 (Pt 4): 697-702.
Pucci, M.J.; Vedamuthu, E.R.; Kunka, B.S. and Vandenbergh, P.A. (1988). Inhibition of Listeria monocytogenes by using bacteriocin PA-1 produced by Pediococcus acidilactici PAC 1.0. Appl. Environ. Microbiol, 54: 2349-53.
Ramesh, A.; Halami, P.M. and Chandrashekar, A. (2000). Ascorbic acid-induced loss of a pediocin-encoding plasmid in Pediococcus acidilactici CFR K7. World Journal of Microbiology and Biotechnology, 16: 695-697.
Ray, S.K.; Johnson, M.C. and Ray, B. (1989). Bacteriocin plasmids of Pediococcus acidilactici. Journal of industrial microbiology, 4: 163-171.
Schved, F.; Lalazar, A.; Henis, Y. and Juven, B.J. (1993). Purification, partial characterization and plasmid-linkage of pediocin SJ-1, a bacteriocin produced by Pediococcus acidilactici. J. Appl. Bacteriol, 74: 67-77.Shamberger, R.J. (1984). Genetic toxicology of ascorbic acid. Mutat Res, 133: 135-59.