• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.143-152
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• 2013

Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system of animals, has several physiological effects including anti-hypertensive, diuretic, tranquilizing, and anti-stress properties, in humans. The purpose of this study was to investigate Lactobacillus plantarum K74, which was isolated from kimchi and selected as a strain with a high ability to produce GABA, to develop a new starter culture for fermented milk production. L. plantarum K74 produced $134.52{\mu}g/mL$ GABA in MRS broth containing 1% MSG, $212.27{\mu}g/mL$ GABA in MRS broth containing 2% MSG, and $234.63{\mu}g/mL$ GABA in MRS broth containing 3% MSG. The optimum growth temperature of L. plantarum K74 was $34^{\circ}C$, reaching a pH of 4.4 after 18 hours of growth. L. plantarum K74 was most sensitive to novobiocin out of 16 different antibiotics tested, and was most resistant to kanamycin and polymyxin B. L. plantarum K74 did not produce ${\beta}$-glucuronidase, a carcinogenic enzyme, and was comparatively tolerant to bile juice and low pH. Furthermore, it displayed resistance to Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus at rates of 54.9%, 46.3%, and 0.7%, respectively.

• Food Science of Animal Resources
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• v.29 no.1
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• pp.15-23
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• 2009

In order to develop a new starter for fermented milk, 2082 bacteria were isolated from raw milk. The strain that showed excellent acid forming and ${\gamma}$-aminobutyric acid (GABA) production ($711.40{\mu}g/g$ D.W) characteristics after incubation at $37^{\circ}C$ for 18 hr was selected and identified as Lactobacillus acidophilus by the result of API carbohydrate fermentation pattern and 16S rDNA sequence. L. acidophilus RMK567 was investigated for its physiological characteristics. RMK67 strain showed good GABA production compared with commercial lactic acid bacteria. The optimum growth temperature of L. acidophilus RMK567 was $40^{\circ}C$ and cultures took 15 hr to reach pH 4.3. L. acidophilus RMK567 showed higher sensitivity to penicillin-G, novobiocin, as compared to other 14 different antibiotics. However, it showed more resistance to kanamycin, neomycin, streptomycin. It showed higher leucine arylamidase and ${\beta}$-galactosidase activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed resistence to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 29.2%, 39.1% and 51.4%, respectively. Based on these and previous results, L. acidophilus RMK567 could be an excellent starter culture for fermented milk with excellent GABA contents.

• Food Science of Animal Resources
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• v.28 no.5
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• pp.587-595
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• 2008

In order to develop a new starter for fermented milk, 1037 bacterial strains were isolated from raw milk. The strain that showed excellent acid producing and angiotensin converting enzyme (ACE) inhibitory activity (88.6%) was selected and identified as a Lactobacillus zeae based on the result of API carbohydrate fermentation pattern and 16S rDNA sequence. Lactobacillus zeae RMK354 was investigated further to study its physiological characteristics. It showed strong ACE inhibitory activity compared with commercial LAB starters tested. The optimum growth temperature of L. zeae RMK354 was $40^{\circ}C$ and it took 10 hr to reach pH 4.3 under this condition. L. zeae RMK354 showed more sensitive to penicillin-G, bacitracin, novobiocin, in a comparison of 14 different antibiotics, and showed most resistance to polymyxin B and vancomycin. It showed higher esterase and leucine arylamidase activities compared with 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed inhibitory activity against Salmonella Typhimurium with the rate of 60%. Based on these and previous results, L. zeae RMK354 could be an excellent starter culture for fermented milk with high level of ACE inhibitory activity.

• Journal of Life Science
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• v.24 no.1
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• pp.46-53
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• 2014

Lactobacillus sakei 383, which showed the highest GABA content in fermented soycurd, survived in artificial gastric fluid (pH 3.0) up to 3 h, and the survival rate was 88%. L. sakei 383 was tolerant to bile juice during incubation in MRS broth with 0.3% oxgall, and the survival rate was 99%. The survival ratio of L. sakei 383 was high in media containing less than 6% NaCl. L. sakei 383 produced an antibacterial substance against various pathogens, including Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Salmonella typhi. The quality characteristics of soycurd fermented with L. sakei 383 were measured during the fermentation period. The viable cell number reached a peak ($10^{11}CFU/ml$) 36 h after fermentation and then slowly decreased. According to the fermentation time of L. sakei 383, the acidity of soycurd increased and the pH decreased until 12 h, and they were maintained thereafter. The moisture, crude ash, crude protein, crude fat, and crude fiber content was 94.88, 0.22, 2.38, 1.16, and 0.03%, respectively. The content of total and reducing sugar was comparatively higher in the soycurd fermented with L. sakei 383 than in nonfermented soycurd. The essential and nonessential amino acid content was 11.2 and 38.65 mg/100 g.

• Journal of Dairy Science and Biotechnology
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• v.32 no.2
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• pp.147-156
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• 2014

The aim of this study was to investigate the physiological characteristics and anti-obesity effects of E. faecalis MD366 isolated from raw milk. E. faecalis MD366 inhibited lipase activity ($65.0{\pm}0.9%$) and differentiation of 3T3-L1 adipocytes ($27.4{\pm}1.4%$) at a concentration of $100{\mu}g/mL$ ($10^8CFU/g$ E. faecalis MD366). The optimum growth temperature of E. faecalis MD366 was $37^{\circ}C$. Among 16 tested antibiotics, E. faecalis MD366 demonstrated the highest sensitivity to novobiocin and the highest resistance to neomycin, kanamycin, and vancomycin. The strain also showed high acid phosphatase activity. Moreover, E. faecalis was relatively tolerant to bile juice and acid, and displayed high resistance to Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus (80.4%, 60.2%, and 65.4%, respectively). These results demonstrate that E. faecalis MD366 can be potentially used as a probiotic with anti-obesity effects.

• Journal of Dairy Science and Biotechnology
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• v.35 no.4
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• pp.221-231
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• 2017

This study aimed to investigate the physiological characteristics and anti-obesity effects of a newly isolated bacterium, Lactobacillus plantarum K6. L. plantarum K6 showed good ${\alpha}-amylase$ inhibitory activity ($96.78{\pm}3.29%$), ${\alpha}-glucosidase$ inhibitory activity ($92.55{\pm}9.62%$), and lipase inhibitory activity ($85.17{\pm}0.79%$), and the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($27.4{\pm}1.4%$) when present at a concentration of $100{\mu}g/mL$. L. plantarum K6 was isolated from kimchi and its physiological characteristics were investigated. A comparison of the sensitivity of the isolate to 15 different antibiotics showed that L. plantarum K6 is highly sensitive to erythromycin and highly resistant to vancomycin, ampicillin, and polymyxin B. This strain also showed high arylamidase and ${\beta}-galactosidase$ activities. Moreover, it was relatively tolerant to bile acid and low pH, and displayed resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, with rates of 51.8%, 42.4%, 61.6%, and 54.9%, respectively. No bio genic amines were produced. L. plantarum K6 also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG. These results demonstrate that Lactobacillus plantarum K6 has potential as a probiotic with anti-obesity effects.

• Food Science of Animal Resources
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• v.27 no.4
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• pp.501-508
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• 2007

In order to develop a new starter for fermented milk, Lactobacillus plantarum M23 was isolated from raw milk and investigated for physiological characteristics. It showed good tyrosinase inhibitory activity compared with commercial lactic acid bacteria. The optimum growth temperature of Lactobacillus plantarum M23 was $40^{\circ}C$ and cultures took 17 hr to reach pH 4.3. Lactobacillus plantarum M23 showed more sensitivity to Penicillin-G, Oxacillin, Novobiocin, Chloramphenicol in a comparison of 12 different antibiotics, and showed most resistance to Vancomycin. It showed higher leucine arylamidase and ${\beta}-galactosidase$ activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hours. It showed high resistance to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 77.8%, 86.5% and 83.8%, respectively. Based on these and previous results, Lactobacillus plantarum M23 could be an excellent starter culture for fermented milk with high resistance to melanin.