Abstract
Antimicrobial proteinaceous compounds such as bacteriocins produced from Lactobacillus sp. are widely known. They have potential antimicrobial activities towards closely related bacteria and several pathogens associated with food spoilage and hence can be a potential food bio-preservative agent. Bacteriocin production requires optimized process, complex media and well-controlled physical conditions including pH and temperature. A probiotic strain of L. casei LA-1 isolated from mango pickle was used in the present study. The influence of physical parameters viz. temperature (15 ∼ 45°C), pH (4.0 ∼ 7.0), incubation time (up to 48 h) and inoculum size (0.7 ∼ 2.0 O.D) on bacteriocin production was analyzed. The effect of all the parameters was first investigated using the one-factor-at-a-time method (OFAT) to see the significance of these parameters on bacteriocin production and then further optimized by response surface methodology (RSM). Following OFAT analysis, all factors were found to have a significant effect on bacteriocin production. Bacteriocin production of 2,844 AU/mL was obtained at temperature 37°C, pH 6.7 and inoculum size 1.8 O.D at an incubation time of 20 h and it was produced during the stationary phase of growth. Statistical analysis showed that three variables-pH, temperature and incubation time have significant effects on bacteriocin production. RSM proved to be a powerful tool in the optimization of bacteriocin production by L. casei LA-1 with a two-fold increase, giving a production of 4652.15 AU/mL at pH 7.19, temperature 33.3°C and incubation time of 22.2 h.
Similar content being viewed by others
References
Aguilar, A. (1991) Biotechnology of lactic acid bacteria: An European perspective. Food Biotechnol. 5: 323–330.
Stiles, M. E. (1996) Biopreservation by lactic acid bacteria. Anton van Leeuw 70: 331–345.
Marrug, J. D. (1995) Bacteriocins, their role in developing natural products. Food Biotechnol. 5: 305–312.
Holzapfel, W. H., R. Geisen, and U. Schillinger (1995) Biological preservation of foods with reference to protective cultures, bacteriocins and food-grade enzymes. Int. J. Food Microbiol. 24: 343–362.
Holtzel, A., M. G. Ganzle, G. J. Nicholson, W. P. Hammes, and G.. Jung (2000) The first low-molecular-weight antibiotic from lactic acid bacteria: Reutericyclin, a new tetramic acid Angewandte. Chem. Int. Edi. 39: 2766–2768.
Magnusson, J. and J. Schnürer (2001) Lactobacillus coryniformis subsp Coryniformis strain Si3 produces a broad-spectrum proteinaceous antifungal compound. Appl. Environ. Microbiol. 67: 1–5.
Daeschel, M. A. (1993) Applications and interactions of bacteriocins from lactic acid bacteria in foods and beverages. pp. 63–91. In: D. G. Hoover and L. R. Steenson (eds.). Bacteriocins of lactic acid bacteria. Academic Press Inc., NY.
De Vuyst, L. and E. J. Vandamme (1994) Lactic acid bacteria and bacteriocins: their practical importance. pp. 1–11. In: L. de Vuyst and E. J. Vandamme (eds.). Bacteriocins of lactic acid bacteria: Microbiology, genetics and applications. Blackie Academic and Professional, London, UK.
Ray, B. and M. A. Daeschel (1994) Bacteriocins of starter culture bacteria. pp. 133–165. In: V. M. Dillon and R. G. Board (eds.). Natural antimicrobial systems and food preservation. CAB International, Wallingford, Oxfordshire, UK.
Antonio Gálvez, Hikmate Abriouel, Rosario Lucas López, and Nabil Ben Omar (2007) Bacteriocin-based strategies for food biopreservation. Int. J. Food Microbiol. 120: 51–70.
Dodd, H. M. and M. J. Gasson (1994) Bacteriocins of lactic acid bacteria. pp. 211–255. In: M. J. Gasson and W. M. de Vos (eds.). Genetics and biotechnology of lactic acid bacteria. Blackie Academic and Professional, London, UK.
Piard, J. -C. and M. J. Desmazeaud (1992) Inhibiting factors produced by lactic acid bacteria 2 Antibacterial substances and bacteriocins. Lait. 72: 113–142.
Eijsink, V. G. H., L. Axelsson, D. B. Diep, L. S. Håvarstein, H. Holo, and I. F. Nes (2002) Production of class II bacteriocins by lactic acid bacteria; an example of biological warfare and communication. Anton. van Leeuw. 81: 639–654.
Holo, H. and I. F. Nes (2000) Class II antimicrobial peptides from lactic acid bacteria. Biopol. 55: 50–61.
Jack, R. W., J. R. Tagg, and B. Ray (1995) Bacteriocins of Grampositive bacteria. Microbiol. Rev. 59: 171–200.
Klaenhammer, T. R. (1993) Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol. Rev. 12: 39–86.
Leal-Sánchez, M. V., R. Jiménez-Díaz, A. Maldonado-Barragán, A. Garrido-Fernández, and J. L. Ruiz-Barba (2002) Optimization of bacteriocin production by batch fermentation of Lactobacillus plantarum LPCO10. Appl. Environ. Microbiol. 68: 4465–4471.
Cássia Regina Nespolo and Adriano Brandelli (2010) Production of bacteriocin-like substances by lactic acid bacteria isolated from regional ovine cheese. Braz. J. Microbiol. 41: 1009–1018
Beatriz Robredo and Carmen Torres (2000) Bacteriocin production by Lactobacillus salivarius of animal origin. J. Clin. Microbiol. 38: 3908–3909.
Yang, R. and B. Ray (1994) Factors influencing production of bacteriocins by lactic acid bacteria. Food Microbiol. 11: 281–291.
Carolissen-Mackay, V., G. Arendse, and J. W. Hastings (1997) Purification of bacteriocins of lactic acid bacteria: Problems and pointers. Int. J. Food Microbiol. 34: 1–16.
Daba, H., C. Lacroix, J. Huang, and R. E. Simard (1993) Influence of growth conditions on production and activity of mesenterocin 5 by a strain of Leuconostoc mesenteroides. Appl. Microbiol. Biotechnol. 39: 166–173.
De Vuyst, L. and E. J. Vandamme (1991) Microbial manipulation of nisin biosynthesis and fermentation. pp. 397–409. In: G. Jung and H. -G. Sahl (eds.). Nisin and novel lantibiotics. ESCOM Science Publishers, Leiden, The Netherlands.
Kaiser, A. L. and T. J. Montville (1993) The influence of pH and growth rate on production of the bacteriocin, bavaricin MN, in batch and continuous fermentations. J. Appl. Bacteriol. 75: 536–540.
Leroy, F. and L. De Vuyst (1999) Temperature and pH conditions that prevail during fermentation of sausages are optimal for production of the antilisterial bacteriocin sakacin K. Appl. Environ. Microbiol. 6: 974–981.
Møortvedt-Abildgaard, C. I., J. Nissen-Meyer, B. Jelle, B. Grenov, M. Skaugen, and I. F. Nes (1995) Production and pH-dependent bactericidal activity of lactocin S, a lantibiotic from Lactobacillus sake L45. Appl. Environ. Microbiol. 61: 175–179.
Parente, E. and A. Ricciardi (1994) Influence of pH on the production of enterocin 1146 during batch fermentation. Let. Appl. Microbiol. 19: 12–15.
Adinarayana, K., P. Ellaiah, B. Srinivasulu, R. B. Devi, and G. Adinarayana (2003) Response surface methodological approach to optimize the nutritional parameters for neomycin production by Streptomyces marinensis under solid-state fermentation. Proc. Biochem. 38: 1565–1572.
Oh, S., S. Rheem, J. Sim, S. Kim, and Y. Baek (1995) Optimizing conditions for the growth of Lactobacillus casei YIT 9018 in tryptone-glucose medium by using response surface methodology. Appl. Environ. Microbiol. 61: 3809–3814.
Li, C., J. Bai, Z. Cai, and F. Ouyang (2001) Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology. J. biotechnol. 93: 27–34.
Kumar, M., M. Ghosh, and A. Ganguli (2011) Mitogenic response and probiotic characteristics of lactic acid bacteria isolated from indigenously pickled vegetables and fermented beverages. World J. Microbiol. Technol. (In press) DOI 10.1007/s11274-011-0866-4.
Motta, A. S. and A. Brandelli (2002) Characterization of an antibacterial peptide produced by Brevibacterium linens. J. Appl. Microbiol. 92: 63–71.
Kimura, H., T. Sashihara, H. Matsusaki, K. Sonomoto, and A. Ishizaki (1998) Novel bacteriocin of Pediococcus sp ISK-1 isolated from wellaged bed of fermented rice. Bran. Ann. NY Acad. Sci. 864: 345–348.
Myers, R. and R. C. Montgomery (2002) Response surface methodology: PROCESS and product optimization using designed experiments. Wiley, NY.
Kim, M. H., Y. J. Kong, H. Baek, and H. Hyun (2006) Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp GO5. J. Biotechnol. 121: 54–61.
Matsusaki, H., N. Endo, K. Sonomoto, and A. Ishizaki (1996) Lantibiotic nisin Z fermentative production by Lactococcus lactis IO-1: Relationship between production of the lantibiotic and lactate and cell growth. Appl. Microbiol. Biotechnol. 45: 36–41.
Cheigh, C. I., H. J. Choi, H. Park, S. B. Kim, M. C. Kook, T. S. Kim, J. K. Hwang, and Y. R. Pyun (2002) Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi. J. Biotechnol. 95: 225–235.
Parente, E., A. Ricciardi, and G. Addario (1994) Influence of pH on growth and bacteriocin production by Lactococcus lactis subsp. Lactis 140NWC during batch fermentation. Appl. Microbiol. Biotechnol. 41: 388–394.
De Vuyst, L., R. Callewaert, and K. Crabbé (1996) Primary metabolite kinetics of bacteriocin biosynthesis by Lactobacillus amylovorus and evidence for stimulation of bacteriocin production under unfavourable growth conditions. Microbiol. 142: 817–827.
Vignolo, G. M., M. N. Kairuz, A. A. P. Ruiz-Holgado, and G. Oliver (1995) Influence of growth conditions on the production of lactocin 705, a bacteriocin produced by Lactobacillus casei CRL 705. J. Appl. Bacteriol. 78: 5–10.
Krier, F., A. M. Revol-Junelles, and P. Germain (1998) Influence of temperature and pH on production of two bacteriocins by Leuconostoc mesenteroides subsp. mesenteroides FR52 during batch fermentation. Appl. Microbiol. Biotechnol. 50: 359–363.
Pilet, M. F., X. Dousset, R. Barre, G. Novel, M. Desmazeaud, and J. C. Piard (1995) Evidence for two bacteriocins produced by Carnobacterium piscicola and Carnobacterium divergens isolated from fishand active against Listeria monocytogenes. J. Food Prot. 58: 256–262.
Stoffels, G., I. Nes, and A. Guomundsdottir (1992) Isolation and properties of a bacteriocin Carnobacterium piscicola isolated from fish. J. Appl. Bacteriol. 73: 309–316.
Biswas, S. R., P. Ray, M. C. Johnson, and B. Ray (1991) Influence of growth conditions on the production of a Bacteriocin, Pediocin AcH, by Pediococcus acidilactici H. Appl. Environ. Microbiol. 57: 1265–1267.
Hurst, A. (1981) Nisin. In: D. Perlman and A. Laskin (eds.). Advances in Applied Microbiology. Academic Press, NY.
Olson, E. R. (1993) Influence of pH on bacterial gene expression. Mol. Microbiol. 8: 5–14.
Cladera-Olivera, F., G. R. Caron, and A. Brandelli (2004) Bacteriocin production by Bacillus licheniformis P40 in cheese whey using response surface methodology. Biochem. Eng. J. 21: 53–58.
Mataragas, M., J. Metaxopoulos, M. Galiotou, and E. H. Drosinos (2003) Influence of pH and temperature on growth and bacteriocin production by Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442. Meat Sci. 64: 265–271.
Cabo, M. L., M. A. Murado, M. Gonzalez, and L. Pastoriza (2001) Effect of aeration and pH gradient on nisin production A mathematical model. Enz. Microbe Technol. 29: 264–273.
De Vuyst, L. (1995) Nutritional factors affecting nisin production by Lactococcus lactis subsp lactis NIZO 22186 in a synthetic medium. J. Appl. Bacteriol. 78: 28–33.
Motta, A. S. and A. Brandelli (2003) Influence of growth conditions on bacteriocin production by Brevibacterium linens. Appl. Microbiol. Biotechnol. 62: 163–167.
Kim, W. S., R. J. Hall, and N. W. Dunn (1997) The effect of nisin concentration and nutrient depletion on nisin production of Lactococcus lactis. Appl. Microbiol. Biotechnol. 50: 429–433.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kumar, M., Jain, A.K., Ghosh, M. et al. Statistical optimization of physical parameters for enhanced bacteriocin production by L. casei . Biotechnol Bioproc E 17, 606–616 (2012). https://doi.org/10.1007/s12257-011-0631-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-011-0631-4