• Journal of Microbiology and Biotechnology
• /
• v.21 no.8
• /
• pp.838-845
• /
• 2011

The present work describes the identification, purification, and characterization of bile salt hydrolase (BSH) from Bifidobacterium animalis subsp. lactis. The enzyme was purified to electrophoretic homogeneity by hydrophobic chromatography, ion-exchange chromatography and ultrafiltration. SDS-PAGE analysis of putative BSH and gel filtration revealed that the analyzed protein is presumably a tetramer composed of four monomers each of about 35 kDa. The purified enzyme was analyzed by liquid chromatography coupled to LTQ FT ICR mass spectrometry and unambiguously identified as a bile salt hydrolase from B. animalis. The isoelectric point of the studied protein was estimated to be around pH 4.9. The pH optimum of the purified BSH is between 4.7 to 6.5, and the temperature optimum is around 50oC. The BSH of B. animalis could deconjugate all tested bile salts, with clear preference for glycine-conjugated bile salts over taurine-conjugated forms. Genetic analysis of the bsh showed high similarity to the previously sequenced bsh gene from B. animalis and confirmed the usefulness of bile salt hydrolase as a genetic marker for B. animalis identification.

• Food Science and Biotechnology
• /
• v.16 no.4
• /
• pp.580-583
• /
• 2007

Phytase activity was examined with various bifidobacterial strains cultured statically in MRS broth at $37^{\circ}C$ for 48 hr. Seven Bifidobacterium species showed mostly an intracellular phytase activity, though their specific activities were very low. The highest specific activity was found in Bifidobacterium animalis B33 strain, among 7 bifidobacteria tested. The specific activity was highest during the exponential growth phase. Carbohydrates and the concentration of phosphorus sources had an effect on the phytase activity and bacterial growth. Glucose was the most favorable carbohydrate for the phytase activity. Phytate inhibited the cell growth, and phytase activity decreased with increase of phytate concentration. The phytase activity was even higher in the static microaerophilic growth than that in anaerobic state, despite the stimulated growth in anaerobic growth. The optimal pH ranges were comparatively broad, but the optimal temperatures were $50^{\circ}C$ for all tested strains. The phytase activity was most active at pH 6.5 and $50^{\circ}C$ for B. animalis B33 strain.

• Journal of Food Hygiene and Safety
• /
• v.21 no.4
• /
• pp.218-222
• /
• 2006

Microbiological and sensory evaluations of bio soybean paste prepared by sesame leaf and immobilized cells of Bifidobacterium animalis DY 64 were assessed. Bio soybean paste treated with 3.0-5.0% (w/w) of sesame leaf combined with 10% (w/w) immobilized cells increased a consumer acceptance due to enhancing odor and flavor. Aerobic microorganisms in bio soybean paste were significantly (P < 0.05) increased during 15 days of storage and then decreased slightly (P < 0.05) after 30 days of storage at room temperature. Food pathogens of Salmonella spp., Staphylococcus aureus and Escherichia coli were not detected in bio soybean paste during storage. It is concluded that preparation of bio soybean paste using sesame leaf, and immobilized cells of Bifidobacterium animalis DY 64 could be used to industrial application for enhancing consumer acceptance.

• Journal of Animal Science and Technology
• /
• v.63 no.3
• /
• pp.603-613
• /
• 2021

This research improved the growth potential of Bifidobacterium animalis subsp lactis strain JNU306, a commercial medium that is appropriate for large-scale production, in yeast extract, soy peptone, glucose, L-cysteine, and ferrous sulfate. Response surface methodology (RSM) was used to optimize the components of this medium, using a central composite design and subsequent analyses. A second-order polynomial regression model, which was fitted to the data at first, significantly lacked fitness. Thus, through further analyses, the model with linear and quadratic terms plus two-way, three-way, and four-way interactions was selected as the final model. Through this model, the optimized medium composition was found as 2.8791% yeast extract, 2.8030% peptone soy, 0.6196% glucose, 0.2823% L-cysteine, and 0.0055% ferrous sulfate, w/v. This optimized medium ensured that the maximum biomass was no lower than the biomass from the commonly used blood-liver (BL) medium. The application of RSM improved the biomass production of this strain in a more cost-effective way by creating an optimum medium. This result shows that B. animalis subsp lactis JNU306 may be used as a commercial starter culture in manufacturing probiotics, including dairy products.

• The Korean Journal of Food And Nutrition
• /
• v.20 no.1
• /
• pp.34-39
• /
• 2007

Microbiological and sensory evaluations were performed on bio-soybean paste treated with citron and immobilized microorganisms from Bifidobacterium animalis DY 64. Aerobic microorganisms in bio-soybean paste stored at room temperature, initially, significantly increased(p<0.05) during 15 days of storage. However, a subsequent, slight decrease(p<0.05) was observed after 30 days. Food pathogens such as Salmonella spp., Staphylococcus aureus and Escherichia coli were not detected in the bio-soybean paste throughout the storage days. Bio-soybean paste treated with 3.0${\sim}$7.0% citron combined with 10% immobilized microorganisms increased consumer acceptance relating odor and flavor. In conclusion, soybean paste treated with citron and immobilized microorganisms from Bifidobacterium animalis could be used as a viable health food with respect to enhancing consumer acceptance.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.3
• /
• pp.431-437
• /
• 2006

Aero-tolerant microorganisms were isolated from healthy Koreans over the age of 95 years. The microorganisms were then identified based on their morphological and biochemical characteristics and 16S rDNA sequences. The growth properties of the isolated strains were investigated in kimchi. The characteristics of kimchi containing the microorganisms were studied microscopically, physicochemically, and organoleptically. Among 7 aero-tolerant strains, a strain with a 16S rDNA sequence exhibiting 99% homology with Bifidobacterim animalis strain B83 was selected and named B. animalis DY-64. The new strain showed a better acid resistance and salt resistance (p<0.05) than B. animalis ATCC 25527. After 15 days of fermentation in kimchi, the viability of B. animalis DY-64 was about 10%, and the kimchi had a better overall edible quality than conventional kimchi. Thus, it was found that the application of B. animalis DY-64 to kimchi preparation produced a good overall edible quality.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.12
• /
• pp.1714-1723
• /
• 2012

${\beta}$-Glucosidase is necessary for the bioconversion of glycosidic phytochemicals in food. Two Bifidobacterium strains (Bifidobacterium animalis subsp. lactis SH5 and B. animalis subsp. lactis RD68) with relatively high ${\beta}$-glucosidase activities were selected among 46 lactic acid bacteria. A ${\beta}$-glucosidase gene (bbg572) from B. lactis was shotgun cloned, fully sequenced, and analyzed for its transcription start site, structural gene, and deduced transcriptional terminator. The structural gene of bbg572 was 1,383 bp. Based on amino sequence similarities, bbg572 was assigned to family 1 of the glycosyl hydrolases. To overexpress bbg572 in Bifidobacterium, several bifidobacteria expression vectors were constructed by combining several promoters and a terminator sequence from different bifidobacteria. The maximum activity of recombinant Bbg572 was achieved when it was expressed under its own promoter and terminator. Its enzyme activity increased 31-fold compared with those of its parental strains. The optimal pH for Bbg572 was pH 6.0. Bbg572 was stable at $37-40^{\circ}C$. It hydrolyzed isoflavones, quercetins, and disaccharides with various ${\beta}$-glucoside linkages. Bbg572 also converted the ginsenosides Rb1 and Rb2. These results suggest that this new ${\beta}$-glucosidase-positive Bifidobacterium transformant can be utilized for the production of specific aglycone products.

• Journal of Dairy Science and Biotechnology
• /
• v.42 no.1
• /
• pp.9-17
• /
• 2024

This study aimed to evaluate the appropriateness of MRS-C (0.05% L-cystein; pH 5) and BHI-CM (0.05% L-cystein, 0.5% mucin) agars for the selective isolation of bifidobacteria in fecal samples compared to blood-liver-NPNL (BL-NPNL) agar. Over 200 isolated colonies were characterized morphologically and biochemically. Genomic DNA was extracted from pure cultures of the isolated strains, followed by PCR amplification of the 16S rRNA gene. Bifidobacterium longum and B. animalis were selectively isolated from MRS-C agar and Lactobacillus acidophilus and Enterococcus avium were also isolated. B. longum, B. faecale, and B. animalis were isolated from feces on BHI-CM agar; however, different Bacteroides strains (including Bac. fragilis, Bac. kiribbi, Bac. ovatus, Bac. koreensis, and Bac. salyersiae) were also detected. BL-NPNL agar successfully isolated B. longum and Bacillus, while other Bifidobacterium and Bacteroides species could not grow owing to the presence of antibiotics in the medium. The use of antibiotics in a medium can enhance the selectivity; however, antibiotics may inhibit the growth of certain bacteria in a sample. Hence, adjusting pH or adding non-antibiotic nutrients to the medium is more advantageous, than relying on antibiotics.

• Korean Journal of Food Science and Technology
• /
• v.39 no.1
• /
• pp.61-65
• /
• 2007

In order to extend the viability of aerotolerant Bifidobacterium animalis DY-64, fructooligosaccharide was added to kimchi containing the bifidobacteria. Baechu-kimchi made with Chinese cabbage was prepared with B. animalis DY-64 and fructooligosaccharide. Physicochemical and microbial changes of the kimchi were evaluated during fermentation at $4^{\circ}C$. Bifidobacteria survived longer in kimchi containing fructooligosaccharide than in kimchi not containing the oligosaccharide. The viable cell counts of Lactobacillus spp. and Leuconostoc spp. and the organic acid content of fructooligosaccharide-added kimchi were higher than those of bifidobacteria or conventional kimchi. The sour taste and sourness of fructooligosaccharide-added kimchi were as high as that of conventional kimchi. These results show that the addition of prebiotic fructooligosaccharide in kimchi enhanced the viability of bifidobacteria during functional kimchi fermentation.

• Journal of Microbiology and Biotechnology
• /
• v.32 no.9
• /
• pp.1146-1153
• /
• 2022

Many probiotic species have been used as a fermentation starter for manufacturing functional food materials. We have isolated Bifidobacterium animalis subsp. lactis LDTM 8102 from the feces of infants as a novel strain for fermentation. While Glycine max has been known to display various bioactivities including anti-oxidant, anti-skin aging, and anti-cancer effects, the immune-modulatory effect of Glycine max has not been reported. In the current study, we have discovered that the extract of Glycine max fermented with B. animalis subsp. lactis LDTM 8102 (GFB 8102), could exert immuno-modulatory properties. GFB 8102 treatment increased the production of immune-stimulatory cytokines in RAW264.7 macrophages without any noticeable cytotoxicity. Analysis of the molecular mechanism revealed that GFB 8102 could upregulate MAPK2K and MAPK signaling pathways including ERK, p38, and JNK. GFB 8102 also increased the proliferation rate of splenocytes isolated from mice. In an animal study, administration of GFB 8102 partially recovered cyclophosphamide-mediated reduction in thymus and spleen weight. Moreover, splenocytes from the GFB 8102-treated group exhibited increased TNF-α, IL-6, and IL-1β production. Based on these findings, GFB 8102 could be a promising functional food material for enhancing immune function.