Production of lactobacillus salivarius, a new probiotic strain isolated from human breast milk, in semi-industrial scale and studies on its functional characterization

Probiotic are living microorganisms when applied to humans or animals, beneficially affect the health of the host by influencing activity of microflora of the gastrointestinal tract or indigineous microbial balance. For industrial production of probiotics belong to lactobacilli, it is neccessary to obtain high biomass in a short time and low cost. Seven media screened for effective production of high biomass were evaluated using shake flask and incubated at 37c, pH 7.0. The best medium supports high biomass and low lactic acid was further used to optimized using different concentrations of medium components. Furthermore, the optimized medium was used for batch cultivation of L. salivarius in bioreactor under controlled and uncontrolled pH conditions. This medium was composed of (g L-1): glucose, 20.0, yeast extract, 20 and meat peptone, 35: Data showed that L. salivarius grew well in this medium with specific growth rate of about 0.179h-1 in shake flask, and 0.249 h-1 in controlled pH bioreactor. The maxiamum biomass of 5.71 g L-1 and 7.57 g L-1, were obtained in shake flasks and controlled pH bioreactor, respectively. To evaluate the potential use of this lactobacilli strain as probiotics, studies on the effect of gastric juice, pH and bile salts were conducted. Cell tolerancey to acidity and bile salt are important factor that affect the probiotics to remain and exert their potential functionalities in a host. L. salivarius showed higher resistant to SGJ with cell viability of 22.9%, 38.8%, 63% and 65% at pH 1,2,3 and 4, respectively. L. salivarius also has good functionality because of its tolerant to wide range bile salt concentrations ranged from 0.5% to 4%. Moreover, L. salivarius was susceptible to antibiotics like erythromycin, rifampicin, ampicillin, and resistance to streptomycin and gentamycin. In conclusion, L. salivarius, new isolated from mother milk, has a big potential use as starter culture probiotic application based on its high stability and could has potential use especially as probiotic supplement for infant milk formulation.

[1]  D. Im,et al.  Screening and characterization of probiotic lactic acid bacteria isolated from Korean fermented foods. , 2009, Journal of microbiology and biotechnology.

[2]  C. Lim,et al.  Optimization of Growth Medium for Efficient Cultivation of Lactobacillus salivarius i 24 using Response Surface Method , 2007 .

[3]  M. Modesto,et al.  Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp. isolated from dairy and pharmaceutical products. , 2007, International journal of food microbiology.

[4]  Yong-Ha Park,et al.  Probiotic properties of Lactobacillus and Bifidobacterium strains isolated from porcine gastrointestinal tract , 2007, Applied Microbiology and Biotechnology.

[5]  Tianwei Tan,et al.  L‐Lactic acid production by Lactobacillus casei fermentation with corn steep liquor‐supplemented acid‐hydrolysate of soybean meal , 2006, Biotechnology journal.

[6]  J. Xaus,et al.  Lactobacillus salivarius CECT 5713, a potential probiotic strain isolated from infant feces and breast milk of a mother-child pair. , 2006, International journal of food microbiology.

[7]  Z. Weizman,et al.  Safety and Tolerance of a Probiotic Formula in Early Infancy Comparing Two Probiotic Agents: A Pilot Study , 2006, Journal of the American College of Nutrition.

[8]  M. Tang,et al.  Probiotic use in clinical practice: what are the risks? , 2006, The American journal of clinical nutrition.

[9]  Peter Neubauer,et al.  Efficient lactic acid production from high salt containing dairy by-products by Lactobacillus salivarius ssp. salicinius with pre-treatment by proteolytic microorganisms. , 2005, Journal of biotechnology.

[10]  L. Vuyst,et al.  Cell growth and bacteriocin production of probiotic Lactobacillus strains in different media , 2004 .

[11]  P. Saris,et al.  Inhibition of Staphylococcus aureus by the commensal bacteria of human milk , 2003, Journal of applied microbiology.

[12]  A. A. de Ruiz Holgado,et al.  Influence of pH, temperature and culture media on the growth and bacteriocin production by vaginal Lactobacillus salivarius CRL 1328 , 2002, Journal of applied microbiology.

[13]  R. Hosken,et al.  Growth medium for culturing probiotic bacteria for applications in vegetarian food products , 2002 .

[14]  S. Yamazaki,et al.  Probiotics and safety. , 2001, The American journal of clinical nutrition.

[15]  B. Kos,et al.  The effect of bile salts on survival and morphology of a potential probiotic strain Lactobacillus acidophilus M92 , 2000 .

[16]  S. Petry,et al.  Factors Affecting Exocellular Polysaccharide Production by Lactobacillus delbrueckii subsp.bulgaricus Grown in a Chemically Defined Medium , 2000, Applied and Environmental Microbiology.

[17]  W. D. de Vos,et al.  Demonstration of safety of probiotics -- a review. , 1998, International journal of food microbiology.

[18]  L. Morelli,et al.  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 , 1998, Journal of applied microbiology.

[19]  J. D. de Bont,et al.  Enhancement of Exopolysaccharide Production byLactobacillus delbrueckii subsp. bulgaricus NCFB 2772 with a Simplified Defined Medium , 1998, Applied and Environmental Microbiology.

[20]  S. Ibrahim,et al.  Survival of bifidobacteria in the presence of bile salt , 1993 .

[21]  B. Goldin,et al.  Probiotics for humans , 1992 .

[22]  A. Bauer,et al.  Antibiotic susceptibility testing by a standardized single disk method. , 1966, American journal of clinical pathology.