• Journal of Biomedical and Translational Research
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• v.19 no.4
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• pp.65-72
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• 2018

The purpose of this study was to investigate probiotic characteristics and immune enhancing effects of Bifidobacterium (B.) longum KBB1-26 and BIF-4, B. breve KBB5-22 isolated from human intestine for probiotic use in humans and animals. We measured acid, bile and heat tolerance, antimicrobial activity against pathogenic bacteria, Escherichia (E.) coli, Salmonella (S.) Enteritidis, Staphylococcus (S.) aureus, and Listeria (L.) monocytogenes. Immune enhancing effects of B. longum and B. breve were investigated by measuring nitric oxide (NO), nuclear factor ($NF-{\kappa}b$), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6), interleukin-12 (IL-12) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in RAW 264.7 cells or RAW BLUE cells. B. longum KBB1-26 was survived at pH 2.0. B. longum KBB1-26 and BIF-4, B. breve KBB5-22 also showed tolerance to 0.3% of oxgall bile salt. B. longum KBB1-26 was able to survive at $70^{\circ}C$ and $80^{\circ}C$ for 20 min. KBB1-26 showed the antimicrobial inhibition zone to pathogenic bacteria such as E. coli (12 mm), S. Enteritidis (14 mm), S. aureus (14 mm) and L. monocytogenes (41 mm). The production of NO ($4.5{\pm}0.00{\mu}M/mL$) and $IL-1{\beta}$ ($39.7{\pm}0.55pg/mL$) of KBB1-26 significantly higher than BIF-4 and KBB5-22, respectively. In addition, KBB1-26 and KBB5-22 induce the production of high level of $TNF-{\alpha}$ and IL-6 in macrophages. Collectively, B. longum KBB1-26 have acid, bile, heat tolerance, antimicrobial activity and immune enhancing effects. These results suggest that KBB1-26 can be used as probiotics for humans and animals.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2100-2105
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• 2015

Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1244-1247
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• 2003

The purpose of this study was to isolate lactic acid bacteria that produced L(+) lactic acid from infant feces. Thirteen colonies were isolated with a MRS-plate containing 0.5% $CaCO_3$ to determine their ability to produce lactic acid. Based on their lactic acid production, 10 strains of Lactobacillus were identified to assess the ratio of lactate isomer using HPLC. A strain producing L-lactic acid was identified as Lactobacillus acidophilus, using API carbohydrate fermentation patterns and physiological tests, and named KY1909. The strain exhibited good acid tolerance in an artificial gastric juice as well as high bile resistance in MRS containing 0.5% bile acids. L. acidophilus KY1909 produced D(-) and L(+) lactic acid at a ratio of 6 : 94; whereas commercial strains of Lactobacillus acidophilus produced D(-) and L(+) lactic acid at a ratio of 1 : 1. These results demonstrate the L. acidophilus KY1909 can be utilized in fermented milk products and dietary supplements as a probiotic culture.

• Food Science of Animal Resources
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• v.24 no.2
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• pp.164-170
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• 2004

The purpose of this study is to isolate probiotic lactic acid bacteria (LAB) that produced bile salts hydrolase. One hundred twenty strains were initially isolated from human feces. Based on their resistance of acid, tolerances of bile salts, and inhibitory activity against Escherichia coli, five strains were selected. A strain producing highest activity of bile salts hydrolase was identified as Lactoacillus plantarum using API carbohydrate fermentation pattern and 16S rRNA sequences, and named CK102. Lactobacillus plantarum CK102 survived at a level of 1.36${\times}$10$\^$8/ CFU/$m\ell$ in pH 2 buffer for 6 h and showed exhibited excellent bile tolerance. When L plantarum CK102 was cultured with E. coli in MRS broth, no viable cells of E. coli was detected after 18 h fermentation. These results suggest that Lactobacillus plantarum CK 102 may be commercially used for the probiotic culture.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.162-165
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• 2002

With the objective of identifying the commercial potential of new direct-fed microbials, several temperature-tolerant strains were isolated from cane molasses at $39^{\circ}C$ and tested for their tolerance to pH, bile salts, and a mixture of volatile fatty acids (acetic acid:propionic acid:butyric acid=6.5:2.0:1.5). It was found that the isolated strain DY 252 grew very well up to pH 2.0 and was resistant to relatively high concentrations of bile salts. Among the strains tested, DY 252 was least inhibited by the addition of volatile fatty acids to the growth medium at $39^{\circ}C$. Accordingly, it would appear that strain DY 252, identified as yeast Issatchenkia orientalis, may be a potential candidate for use as a microbial feed additive.

• KSBB Journal
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• v.14 no.4
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• pp.389-395
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• 1999

Bifidobcterium spp. can provide human being with several beneficial physiological. Therefor, there has been a considerable interest in products Bifidobcterium spp. dietary supplements or as starter cultures for probiotic products that may assint in the improvement of health on the human. But indusrial applications have been limited because Bifidobcterium spp. are sensitive to acidic pH due to organic acid produced by themselves and various conditions. The objective of this study was to establish new method for improvement of Bifidobcterium viability by entrapment im calcium alginate beads. We have a plan to select the most suitable polymer through the comparison with acid tolerance oxygen tolerance and theological properties of polymer. Increase of the viable number of Bifidobcterium induced increasing acid tolerance and oxygen tolernce trough the development of entrapment technique. The 4%, 3030mm diameter) sodium alginate beads led to the best survivability under acid condition. Especially, addition of 6% mannitol, 6% glycerol or 6% sorbitol to the sodium alginate helped a beneficial effect on viability against acid, bile salt, hydrogen peroxide and cold strage. The number of viability of entrapeede cells by retreatment was 96 fold higher than non-entrapeed cells after 5 hours of storage under pH 3 acidic condition. These experimental data clearly demonstrate that a whole cell immobilization by entrapment in calcium alginate beads is an important survival mechanism enable to withstand environmental stresses as the acidic condition, hydrogen peroxide toxicity and frozen state.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.276-286
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• 2020

Probiotics are live microorganisms that, when administered in adequate amounts, confer health benefits to the host. This study was conducted for the isolation of potential lactic acid bacteria (LAB) with probiotic properties from goat milk and yogurt. Several tests were conducted in vitro using the standard procedures for evaluating the inhibitory spectra of LAB against pathogenic bacteria; tolerance to NaCl, bile salt, and phenol; hemolytic, milk coagulation, and bile salt hydrolase activities; gastrointestinal transit tolerance; adhesion properties; and antibiotic susceptibility. Among 40 LAB strains screened according to culture characteristics, five isolates exhibited antagonistic properties. Three were identified as Pediococcus acidilactici, and two were identified as Enterococcus faecium, exploiting 16S rRNA gene sequencing. All the isolates succeeded in the gastrointestinal transit tolerance assay and successively colonized mucosal epithelial cells. Based on the results of these in vitro assays, both P. acidilactici and E. faecium can be considered as potential probiotic candidates.

• Korean Journal of Food Science and Technology
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• v.17 no.3
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• pp.192-196
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• 1985

Eight strains of Lactobacilli(a, b, b', c, d, e, f and g) were isolated from seven Korean liquid-yogurts(A, B, C, D, E, F and G), and identification and tolerance-test to digestive fluids were carried out. Isolate a from yogurt A and isolate a from yogurt E were identified as L. casei, isolate b from yogurt B as L. acidophilus, isolate d from yogurt D as L. bulgaricus, isolate f from yogurt F as L. helveticus, and isolate b' from yogurt B, isolate c from yogurt C and isolate g from yogurt G as L. jugurti, respectively. Isolate f(L. helveticus) and c(L. jugurti) showed high tolerance to artificial gastric juice but didn't to bile acid. Isolate b(L. acidophilus), a(L. casei), and e(L. casei) showed high tolerance to both artificial gastric juice and bile acid, but isolate d(L. bulgaricus), b'(L. jurgurti) and g(L. jugurti) did not.

• KSBB Journal
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• v.20 no.2 s.91
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• pp.76-83
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• 2005

Lactic acid bacteria were isolated from pig feces for probiotics. The six isolated strains were identified as Lactobacillus paracasei (Lp), Lactobacillus fermentum (Lf), Lactobacillus brevis (Lb), Lactobacillus plantarum (P1 , P2), and Pediococcus pentosaceus (P3) by its sugar utilization, morphological and physiological characteristics. Pl was showed largest antibacterial inhibition zone among the isolated strains. It was against Salmonella gallinarum 25mm, E. coli 20.5mm, Staphylococcus aures 24mm, and Pseudomonas aeruginosa 28mm by inhibitory zone, respectively. Lf was showed hyper acid tolerance, $80\%$ survival rate for 40 minutes, and P1, Lb showed hyper bile tolerance, $408\%,\;283\%$ survival rate for 9 hrs, respectively. Therefore the Lf, P1, and P2 strains were expected to probiotics.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.783-791
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• 2022

Gamma-aminobutyric acid (GABA) improves various physiological illnesses, including diabetes, hypertension, depression, memory lapse, and insomnia in humans. Therefore, interest in the commercial production of GABA is steadily increasing. Lactic acid bacteria (LAB) have widely been reported as a GABA producer and are safe for human consumption. In this study, GABA-producing LAB were preliminarily identified and quantified via GABase assay. The acid and bile tolerance of the L. plantarum FBT215 strain were evaluated. The one-factor-at-a-time (OFAT) strategy was applied to determine the optimal conditions for GABA production using HPLC. Response surface methodology (RSM) with Box-Behnken design was used to predict the optimum GABA production. The strain FBT215 was shown to be acid and bile tolerant. The optimization of GABA production via the OFAT strategy resulted in an average GABA concentration of 1688.65 ± 14.29 ㎍/ml, while it was 1812.16 ± 23.16 ㎍/ml when RSM was applied. In conclusion, this study provides the optimum culture conditions for GABA production by the strain FBT215 and indicates that L. plantarum FBT215 is potentially promising for commercial functional probiotics with health claims.