Journal of Food Science and Technology

, Volume 47, Issue 3, pp 258–265 | Cite as

Simplification and optimization of deMan Rogosa Sharpe (MRS) medium for enhanced production of bacteriocin by Weissella paramesenteroides DFR-8

  • Ajay Pal
  • K. V. Ramana
  • A. S. Bawa
Original Article


Complex growth medium such as deMan Rogosa Sharpe (MRS) medium, commonly used for cultivation of fastidious lactic acid bacteria (LAB) interfere in bacteriocin purification. Sometimes all the ingredients of a defined medium are not required by all LAB strains for bacteriocin production. In the present study, composition of the MRS medium for the production of bacteriocin by Weissella paramesenteroides DFR-8, an isolate from cucumber (Cucumis sativus), was simplified and optimized with a step-wise strategy. In the first step, production profile, effect of incubation temperature, various C and N sources were investigated. In the second step, central composite rotatable design was employed to decide the optimal concentration of 3 key components (glucose, tryptone and pH) and the experimental results were fitted with a second order polynomial regression equation. According to the set criteria, the predicted bacteriocin titer from a medium containing 7.99% glucose, 9% tryptone, pH 7.5 (91.9% desirability) was 540 AU/ml and the observed bacteriocin titer was 538 AU/ml that indicated the validity of the developed model. Using optimized medium, bacteriocin titer of 674.5 AU/ml could be achieved after 72 h of fermentation that is nearly 2.5 fold higher than that obtained from unmodified MRS medium.


Bacteriocin Biopreservation Optimization Weissella paramesenteroides 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adinarayana K, Ellaiah P, Srinivasulu B, Devi RB, Adinarayana G (2003) Response surface methodological approach to optimize the nutritional parameters for neomycin production by Streptomyces marinensis under solid-state fermentation. Process Biochem 38:1565–1572CrossRefGoogle Scholar
  2. Cabo ML, Murado MA, Gonzalez M, Pastoriza L (2001) Effects of aeration and pH gradient on nisin production, a mathematical model. Enz Microbial Technol 29:264–273CrossRefGoogle Scholar
  3. Cheigh CI, Choi HJ, Park H, Sim SB, Kook MC, Kim TS, Hwang JK, Pyun YR (2002) Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi. J Biotechnol 95:225–235CrossRefGoogle Scholar
  4. Delgado A, Lopez FN, Brito D, Peres C, Fevereiro P, Fernandez AG (2007) Optimum bacteriocin production by Lactobacillus plantarum 17.2b requires absence of NaCl and apparently follows a mixed metabolite kinetics. J Biotechnol 130:193–201CrossRefGoogle Scholar
  5. de Vuyst L (1995) Nutritional factors affecting nisin production by Lactococcus lactis subsp. lactis NIZ022186 in a synthetic medium. J Appl Bacteriol 78:28–33Google Scholar
  6. de Vuyst L, Callewaert R, Crabbe K (1996) Primary metabolite kinetics of bacteriocin biosynthesis by Lactobacillus amylovorus and evidence for stimulation of bacteriocin production under unfavorable growth condition. Microbiol 142:817–827CrossRefGoogle Scholar
  7. Dominguez APM, Bizani D, Cladera-Olivera F, Brandelli A (2007) Cerein 8A production in soybean protein using response surface methodology. Biochem Eng J 35:238–243CrossRefGoogle Scholar
  8. Jamuna M, Jeevaratnam K (2004) Isolation and characterization o lactobacilli from some traditional fermented foods and evaluation of their bacteriocins. J Gen Appl Microbiol 50:79–90CrossRefGoogle Scholar
  9. Joshi MS, Gowda LR, Bhat SG (1987) Permeabilization of yeast cells (Kluyveromyces fragilis). Biotechnol Lett 9:549–554CrossRefGoogle Scholar
  10. Kim CH, Ji GE, Ahn C (2000) Purification and molecular characterization of a bacteriocin from Pediococcus sp. KCA 1303-10 isolated from fermented flat fish. Food Sci Biotechnol 19:270–276Google Scholar
  11. Kim MC, Kong YJ, Baek H, Hyun HH (2006) Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp. GO5. J Biotechnol 21:54–61CrossRefGoogle Scholar
  12. Li C, Bai J, Cai Z, Ouyang F (2001) Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology. J Biotechnol 93:27–34CrossRefGoogle Scholar
  13. Mackay VC, Arendse G, Hastings JW (1997) Purification of bacteriocins of lactic acid bacteria: problems and pointers. Int J Food Microbiol 34:1–16CrossRefGoogle Scholar
  14. Matsusaki H, Endo N, Sonomoto K, Ishizaki A (1996) Lantibiotic nisin Z fermentative production by Lactococcus lactis 10-1: Relationship between production of the lantibiotic and lactate and cell growth. Appl Microbiol Biotechnol 45:36–40CrossRefGoogle Scholar
  15. Motta AS, Brandelli A (2003) Influence of growth conditions on bacteriocin production by Brevibacterium linens. Appl Microbiol Biotechnol 62:163–168CrossRefGoogle Scholar
  16. Myers RH, Montogomery RC (2002) Response surface methodology: process and product optimization using designed experiments. Wiley, New YorkGoogle Scholar
  17. Oh S, Rheem S, Sim J, Kim S, Back Y (1995) Optimizing conditions for the growth of Lactobacillus casei YIT 9018 in tryptone-glucose medium by using response surface methodology. Appl Environ Microbiol 61:3809–3814Google Scholar
  18. Pal A, Ramana KV (2009a) Isolation and preliminary characterization of a nonbacteriocin antimicrobial compound from Weissella paramesenteroides DFR-8 isolated from cucumber (Cucumis sativus). Process Biochem 44:499–503CrossRefGoogle Scholar
  19. Pal A, Ramana KV (2009b) Purification and characterization of bacteriocin from Weissella paramesenteroides DFR-8, an isolate from cucumber (Cucumis sativus). J Food Biochem (in press, jfbc-07-08-0183, Date of acceptance 2 Dec 2008)Google Scholar
  20. Puri S, Beg QK, Gupta R (2001) Optimization of alkaline protease production from Bacillus sp. by response surface methodology. Current Microbiol 44:286–290CrossRefGoogle Scholar
  21. Sudha N, Rani SP, Agarwal R (2006) Studies on the stability and viability of a local probiotic isolate Pediococcus pentosaceous (MTCC 5151) under induced gastrointestinal tract conditions. J Food Sci Technol 43:677–678Google Scholar
  22. Tagg JR, McGiven AR (1971) Assay system for bacteriocins. Appl Microbiol 21:125Google Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2010

Authors and Affiliations

  1. 1.Food Biotechnology DisciplineDefence Food Research LaboratoryMysoreIndia

Personalised recommendations