Current Science

Open Access Open Access  Restricted Access Subscription Access

Bioprocess Development for Enhanced Production of Probiotic Bifidobacterium bifidum


Affiliations
1 Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India
 

The objective of this study was the development of bioprocess for enhanced biomass production of probiotic Bifidobacterium bifidum. In the first process optimization step in Erlenmeyer flasks cultures, different experiments were conducted to study the effect of inoculum volume, inoculum age, temperature and pH of the growth medium on the kinetics of cell growth. In Erlenmeyer flasks cultures, the maximal biomass production was observed with 1% inoculum of 6 log hours at 37°C, and optimal pH of initial media was found to be 6.0. Further positive development in biomass production was observed by scaling up the fermentation process to stirred tank bioreactor. Fermentation was carried out in 2L stirred tank bioreactor, with agitation of 100 rpm and constant temperature of 37°C. The batch culture produced higher biomass of 34.1 g wet cell weight g/l in 12 log hours and viable counts (2.5 × 109 CFU/ml) compared to Erlenmeyer flasks. In conclusion, batch cultivation in the 2 l bioreactor with this growth medium under optimal conditions gives enhanced biomass production. However, based on our end result, high-cell density fed-batch and pH control strategies are recommended for the commercial production of B. bifidum as a probiotic.

Keywords

Bifidobacterium bifidum, Bioprocess Development, Culture Condition, Probiotics.
User
Notifications
Font Size

  • Bull, M. J. and Plummer, N. T., Part 1: the human gut microbiome in health and disease. Integr. Med., 2014, 17(6), 17.

  • Gibson, G. R. and Roberfroid, M. B., Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J. Nutr., 1995, 125(6), 1401–1412.

  • Sartor, R. B. and Mazmanian, S. K., Intestinal microbes in inflammatory bowel diseases. Am. J. Gastroenterol., 2012, 1(1), 15.

  • Food and Agriculture Organization and World Health Organization. Probiotics in food: health and nutritional properties and guidelines for evaluation. FAO, Rome, 2006.

  • Santiesteban-López, A., López-Malo, J. J., Gomez-Diaz, B. and Armendariz, P., Effect of L-cysteine and ascorbic acid on the propagation of Lactobacillus casei in milk. Int. Res. J. Microbiol., 2013, 4, 113–118.

  • Oak, S. J. and Jha, R., The effects of probiotics in lactose intolerance: a systematic review. Crit. Rev. Food Sci. Nutr., 2018, 59(11), 1675–1683.

  • Gismondo, M. R., Drago, L. and Lombardi, A., Review of probiotics available to modify gastrointestinal flora. Int. J. Antimicrob. Agents, 1999, 12(4), 287–292.

  • Malyoth, G. and Bauer, A., Observations on Bacterium bifidum. Z. Kinderhelikd., 1950, 68(4), 358.

  • Culpepper, T. et al., Bifidobacterium bifidum R0071 decreases stress associated diarrhoea-related symptoms and self-reported stress: a secondary analysis of a randomised trial. Benef. Microbes, 2016, 7(3), 327–336.

  • Saarela, M., Mogensen, G., Fonden, R., Mättö, J. and MattilaSandholm, T., Probiotic bacteria: safety, functional and technological properties. J. Biotechnol., 2000, 84(3), 197–215.

  • Siaterlis, A., Deepika, G. and Charalampopoulos, D., Effect of culture medium and cryoprotectants on the growth and survival of probiotic lactobacilli during freeze drying. Lett. Appl. Microbiol., 2009, 48, 295–301.

  • Veda, M. O., Nakamoto, S., Nakai, R. and Takagi, A., Establishment of a defined minimal medium and isolation of auxotrophic mutants for Bifidobacterium bifidum ES 5. J. Gen. Appl. Microbiol., 1983, 29(2), 103–114.

  • Gaden Jr, E. L., Fermentation process kinetics. Biotechnol. Bioeng., 2000, 67(6), 629–635.

  • Corre, C., Madec, M. N. and Boyaval, P., Production of concentrated Bifidobacterium bifidum. J. Chem. Technol. Biotechnol., 1992, 53(2), 189–194.

  • De Man, J. C., Rogosa, M. and Sharpe, M. E., A medium for the cultivation of lactobacilli. J. Appl. Bacteriol., 1960, 23, 130–135.

  • Vinderola, C. G. and Reinheimer, J. A., Culture media for the enumeration of Bifidobacterium bifidum and Lactobacillus acidophilus in the presence of yoghurt bacteria. Int. Dairy J., 1999, 9(8), 497–505.

  • Coghetto, C. C., Brinques, G. B., Siqueira, N. M., Pletsch, J., Soares, R. M. and Ayub, M. A., Electrospraying microencapsulation of Lactobacillus plantarum enhances cell viability under refrigeration storage and simulated gastric and intestinal fluids. J. Funct. Foods, 2016, 24, 316–326.

  • Meena, G. S., Gupta, S., Majumdar, G. C. and Banerjee, R., Growth characteristics modeling of Bifidobacterium bifidum using RSM and ANN. Braz. Arch. Biol. Technol., 2011, 54(6), 1357– 1366.

  • Dinakar, P. and Mistry, V. V., Growth and viability of Bifidobacterium bifidum in cheddar cheese. J. Dairy Sci., 1994, 77(10), 2854–2864.

  • Hekmat, S., Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in ice cream for use as a probiotic food. J. Dairy Sci., 1992, 6, 1415–1422.

  • Kulkarni, S., Haq, S. F., Samant, S. and Sukumaran, S., Adaptation of Lactobacillus acidophilus to thermal stress yields a thermotolerant variant which also exhibits improved survival at pH 2. Probiot. Antimicrob. Proteins, 2018, 10(4), 717–727.

  • Hwang, C. F., Lin, C. K., Yan, S. Y., Chang, R. H. and Tsen, H. Y., Enhancement of biomass production and nutrition utilization by strain Lactobacillus acidophilus DGK derived from serial subculturing in an aerobic environment. Afr. J. Biotechnol., 2015, 14(3), 248–256.

  • Ram, C. and Chander, H., Optimization of culture conditions of probiotic bifidobacteria for maximal adhesion to hexadecane. World J. Microbiol. Biotechnol., 2003, 19(4), 407–410.

  • Mlobeli, N. T., Batch culture studies of Bifidobacterium bifidum: a thesis presented in partial fulfilment of the requirements for the degree of Master of Technology in Biotechnology and Bioprocess Engineering, Doctoral dissertation, Massey University, 1996.

  • Kwon, S. G., Son, J. W., Kim, H. J., Park, C. S., Lee, J. K., Ji, G. E. and Oh, D. K., High concentration cultivation of Bifidobacterium bifidum in a submerged membrane bioreactor. Biotechnol. Prog., 2006, 22(6), 1591–1597.


Abstract Views: 461

PDF Views: 135




  • Bioprocess Development for Enhanced Production of Probiotic Bifidobacterium bifidum

Abstract Views: 461  |  PDF Views: 135

Authors

Suhail Mohammed Hussain
Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India
Manali Naik
Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India
L. Arbaaz Ahmed
Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India
Minal Udipi
Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India
Sunil Kumar Sukumaran
Anthem Biosciences Pvt Ltd, Bommasandra Industrial Area Phase-I, Hosur Road, Bengaluru 560 099, India

Abstract


The objective of this study was the development of bioprocess for enhanced biomass production of probiotic Bifidobacterium bifidum. In the first process optimization step in Erlenmeyer flasks cultures, different experiments were conducted to study the effect of inoculum volume, inoculum age, temperature and pH of the growth medium on the kinetics of cell growth. In Erlenmeyer flasks cultures, the maximal biomass production was observed with 1% inoculum of 6 log hours at 37°C, and optimal pH of initial media was found to be 6.0. Further positive development in biomass production was observed by scaling up the fermentation process to stirred tank bioreactor. Fermentation was carried out in 2L stirred tank bioreactor, with agitation of 100 rpm and constant temperature of 37°C. The batch culture produced higher biomass of 34.1 g wet cell weight g/l in 12 log hours and viable counts (2.5 × 109 CFU/ml) compared to Erlenmeyer flasks. In conclusion, batch cultivation in the 2 l bioreactor with this growth medium under optimal conditions gives enhanced biomass production. However, based on our end result, high-cell density fed-batch and pH control strategies are recommended for the commercial production of B. bifidum as a probiotic.

Keywords


Bifidobacterium bifidum, Bioprocess Development, Culture Condition, Probiotics.

References





DOI: https://doi.org/10.18520/cs%2Fv118%2Fi2%2F280-285