AiM  Vol.8 No.1 , January 2018
Molecular and Probiotic Characterizations of Lactobacillus reuteri DSM 12246 and Impact of pH on Biomass and Metabolic Profile in Batch-Culture
Abstract: Lactobacillus reuteri is a powerful probiotic and adjunct functional culture candidate received a lot of scientific attention as it is one of the few endogenous “Lactobacillus” species found in the gastrointestinal tract of vertebrates, including humans, rats, pigs and chicken. The organism has been widely utilized as a probiotic in humans and animals for many years. In the present work L. reuteri DSM 12243; a high reuteri producer strain was molecularly characterized by 16SrRNA and RAPD analyses further, the probiotic properties including acid resistance, bile tolerance, adhesion to epithelial gastric cells, and antibiotic susceptibility were also assessed. Furthermore, the effect of pH on biomass production and metabolic profile of L. reuteri DSM 12246 in batch-culture was studied. The L. reuteri DSM 12246 showed a high similarity with L. reuteri strain I49 KR 36477 (100%) and type strain ATCC 55730 (99% of identity). The strain adhered well to CaCO2 cells and showed to be a highly resistance to acid juice (pH 3.0), with 0.7 log10 cfu/ml reduction in count after 60 min exposition. There is no significant change in the cell count after exposure to bile salts. In batch-cultures, at low pH values both glucose consumption and metabolites were low while the production of lactic acid was noticeable. Maximum biomass was reached at pH 5.5, with growth rate of μ = 0.641/h. The switch in pH values from 3.7 to 6.7 resulted in raising of glucose depletion as well as in the yield of acetate and ethanol. It is concluded that L. reuteri DSM 12246 was deemed as a successful candidate to be used as potential probiotic.
Cite this paper: El-Ziney, M. (2018) Molecular and Probiotic Characterizations of Lactobacillus reuteri DSM 12246 and Impact of pH on Biomass and Metabolic Profile in Batch-Culture. Advances in Microbiology, 8, 18-30. doi: 10.4236/aim.2018.81002.

[1]   Britton, R.A. (2017) Lactobacillus reuteri. In: Floch, M.H., Ringel, Y. and Walker, W.A., Eds., The Microbiota in Gastrointestinal Pathophysiology: Implications for Human Health, Prebiotics, Probiotics, and Dysbiosis, Elsevier Inc., Amsterdam, 89-97.

[2]   El-Ziney, M.G. and Debevere, J.M. (1998) The Effect of Reuterin on Listeria monocytogenes and Escherichia coli O157:H7 in Milk and Cottage Cheese. Journal of Food Protection, 10, 1275-1280.

[3]   El-Ziney, M.G., van den Tempel, T., Debevere, J., et al. (1999) Application of Reuterin Produced by Lactobacillus reuteri 12002 for Meat Decontamination and Preservation. Journal of Food Protection, 62, 257-261.

[4]   Martin-Cabrejas, I., Langa, S., Gaya, P., et al. (2017) Optimization of Reuterin Production in Cheese by Lactobacillus reuteri. Journal of Food Science and Technology, 54, 1346-1349.

[5]   Donelli, G., Vuotto, C. and Mastromarino, P. (2013) Phenotyping and Genotyping are Both Essential to Identify and Classify a Probiotic Microorganism. Microbial Ecology Health and Disease, 24, 20105.

[6]   Bongaerts, G. and Severijnen, R. (2007) Probiotics: Are They Incredible Panaceas? On the Science behind Beneficial Non-Pathogenic Microbial Agents. International Journal of Probiotics and Prebiotics, 1, 87-96.

[7]   Goldin, B.R., Gorbach, S.L., Saxelin, M., et al. (1992) Survival of Lactobacillus Species (strain LGG) in Human Gastrointestinal Tract. Digestive Diseases and Sciences, 37, 121-128.

[8]   El-Ziney, M.G., Arneborg, N., Uyttendaele, M., et al. (1998) Characterization of Growth and Metabolite Production of Lactobacillus reuteri during Glucose/Glycerol Cofermentation in Batch and continuous Cultures. Biotechnology Letters, 20, 913-916.

[9]   Fontana, C., Sandro, C.P. and Vignolo, G. (2005) Monitoring the Bacterial Population Dynamics during Fermentation of Artisanal Argentinean Sausages. International Journal Food Microbiology, 103, 131-142.

[10]   Charteris, W.P., Kelly, P.M., Morelli, L., et al. (1998) Development and Application of an In Vitro Methodology to Determine the Transit Tolerance of Potentially Probiotic Lactobacillus and Bifidobacterium Species in the Upper Human Gastrointestinal Tract. Journal Applied Microbiology, 84, 759-768.

[11]   Kaushik, J.K., Kumar, A., Duary, R.K., et al. (2009) Functional and Probiotic Attributes of an Indigenous Isolate of Lactobacillus plantarum. PLoS One, 4.

[12]   EFSA Scientific Opinion (2012) Guidance on the Assessment of Bacterial Susceptibility to Antimicrobials of Human and Veterinary Importance. European Safety Authority Journal, 10, 2740.

[13]   ISO 10932 / IDF 223:2010 (2010) Milk and Milk Products—Determination of the Minimal Inhibitory Concentration (MIC) of Antibiotics Applicable to Bifidobacteria and Non-Enterococcal Lactic Acid Bacteria (LAB).

[14]   Elkins, C.A. and Mullis, L.B. (2004) Bile-Mediated Aminoglycoside Sensitivity in Lactobacillus Species Likely Results from Increased Membrane Permeability Attributable to Cholic Acid. Applied and Environmental Microbiology, 12, 7200-7209.

[15]   Huys, G., D’Haene, K. and Swings, J. (2002) Influence of the Culture Medium on Antibiotic Susceptibility Testing of Food-Associated Lactic Acid Bacteria with the Agar Overlay Disc Diffusion Method. Letters in Applied Microbiology, 34, 402-406.

[16]   Egervärn, M., Roos, S. and Lindmark, H. (2009) Identification and Characterization of Antibiotic Resistance Genes in Lactobacillus reuteri and Lactobacillus plantarum. Journal of Applied Microbiology, 107, 1658-1668.

[17]   Branton, W.B., Jones, M.L., Tomaro-Duchesneau, C., et al. (2011) In Vitro Characterization and Safety of the Probiotic Strain Lactobacillus reuteri Cardioviva NCIMB 30242. International Journal of Probiotics Prebiotics, 6, 1-12.

[18]   Egervärn, M., Lindmark, H., Olsson, H., et al. (2010) Transferability of a Tetracycline Resistance Gene from Probiotic Lactobacillus reuteri to Bacteria in the Gastrointestinal Tract of Humans. Antonie van Leeuwenhoek, 97, 189-200.

[19]   Rosander, A., Connolly, E. and Roos, S. (2008) Removal of Antibiotic Resistance Gene-Carrying Plasmids from Lactobacillus reuteri ATCC 55730 and Characterization of the Resulting Daughter Strain, L. reuteri DSM 17938. Applied and Environmental Microbiology, 74, 6032-6040.

[20]   FDA (2008).

[21]   Kandler, O. (1983) Carbohydrate Metabolism in Lactic Acid Bacteria. Antonie van Leeuwenhoek, 49, 209-224.

[22]   Ragout, A., Siñeriz, F., Diekmann, H., et al. (1994) Effect of Environmental pH on the Fermentation Balance of Lactobacillus reuteri. Journal of Applied Bacteriology, 77, 388-391.

[23]   Dainty, R.H. and Hofman, F.J.K. (1983) The Influence of Glucoseconcentration and Culture Incubation Time on End-Product Formation during Aerobic Growth of Brochothrix thermosphacta. Journal of Applied Bacteriology, 55, 233-239.

[24]   Speck, E.L. and Freese, E. (1973) Control of Metabolite Secretionin Bacillus subtilis. Journal of General Microbiology, 78, 261-275.