• Food Science and Preservation
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• v.24 no.5
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• pp.707-713
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• 2017

The objective of this study was to select yogurt starter from Korean traditional fermented foods. The 2 strains (KM24, KM32) among 50 strains of isolated lactic acid bacteria selected as starter based on milk clotting ability, antimicrobial activity against various pathogens, tolerance in artificial gastric and bile juice and growth in 10 % skimmed milk. The strains were identified as Lacobacillus plantarum (KM32) and Pediococcus pentosacesus (KM24) by 16S rRNA gene sequencing. Viable cell number of yogurt fermented with mixed strains (KM24 and KM32) was 9.66 log CFU/mL after fermentation for 48 h and maintained $10^9CFU/mL$ during fermentation for 72 h at $37^{\circ}C$. The pH and titratable acidity of mixed cultured yogurt were 4.25% and 0.83% after fermentation for 48 h at $37^{\circ}C$, respectively. The physico-chemical characteristics of mixed cultured yogurt after fermentation for 48 h were $38.45{\mu}g/mL$ (polyphenol content), 48.57% (DPPH radical scavenging activity) and 465.40 cp (viscosity), respectively. The mixed cultured yogurt maintained $10^9CFU/mL$ of lactic acid bacteria during storage 10 days at $4^{\circ}C$. The viable cell number of yogurt prepared with mixed culture(KM32+KM24) maintained higher and than that of control (L. casei) during storage. These results indicated the potential use of selected strains (KM32+KM24) isolated from kimchi as a yogurt starter with strong acid tolerance and probiotics properties.

• 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.

• Food Science and Preservation
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• v.23 no.5
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• pp.718-726
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• 2016

Sweet pumpkin paste (SPP) was fermented by Leuconostoc mesenteroides SM at $25^{\circ}C$ for 3 days for enhancing its physicochemical properties. SPPs with 5%, 10%, and 15% solid contents (SC) were fortified with 20% sucrose and 0.5% yeast extract. The unfermented SPP with 15% SC indicated L, a, and b color values of 25.02, 4.66, and 13.35, respectively, and a consistency index of $48.6Pa{\cdot}s^n$. During the 3 days of fermentation, both the a and b color values decreased slightly, whereas the consistency index increased to $188.8Pa{\cdot}s^n$, giving the fermented product a pudding-like consistency. This fermented SPP (15% SC) showed the highest acid production and viable cell counts among samples, indicating pH 3.85, 1.30% acidity and $9.2{\times}10^8CFU/mL$ respectively. The added sucrose was completely utilized after 1 day of fermentation. After 3 days, the insoluble and soluble dextran contents were 8.9% and 4.5%, respectively. Furthermore, the contents of mannitol and fructose were 3.11% and 1.76%, respectively. Regarding the sensory evaluation, this fermented sample also indicated the highest color, taste and texture scores, and was the overall preferred sample. In conclusion, the fermented SPP with 15% SC was carotinoid-rich a wholesome pumpkin-based product that is rich in probiotics and lactic bacteria-produced mannitol and dextran, which gave the product an acceptable viscous pudding-like consistency and good organoleptic properties.

• Food Science and Preservation
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• v.24 no.6
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• pp.865-870
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• 2017

Yogurt is produced by fermentation of milk using bacteria known as "yogurt cultures". Most of these bacteria are probiotics such as Lactobacillus delbrueckii subsp. bulgaricus, Lb. rhamnosus, Streptococcus thermophilus, and Bifidobacterium. The domestic fermented milk market is increasing, and about 30 companies are producing yogurt. The purpose of this study was to analyze the quality characteristics of domestic commercial liquid yogurt. Total 30types of commercial yogurts were sampled and their physicochemicial properties, including pH, sugar content, acidity, viscosity, and microbial characteristics of lactic acid bacteria counts, were measured. Commercial liquid type yogurt showed a pH of 4.5, sugar content of 7.4-21.2%, total acid content of 0.4-0.9%, and viscosity of 0.1-250 cP. In terms of microbial populations, lactic acid bacteria counts were 7.2-11.3 log CFU/mL and anaerobic lactic acid bacteria counts were 8.0-11.5 log CFU/mL. The quality characteristics differed depending on the constituents of the sample and the microorganisms used. These results are related to the quality characteristics of yogurts and are useful for identifying new trends in the domestic fermented milk industry.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1018-1025
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• 2016

Probiotics are considered as the best effective alternatives to antibiotics. The aim of this study was to characterize the probiotic potential of lactobacilli for use in swine farming by using in vitro evaluation methods. A total of 106 lactic acid bacterial isolates, originating from porcine feces, were first screened for the capacity to survive stresses considered important for putative probiotic strains. Sixteen isolates showed notable acid and bile resistance, antibacterial activity, and adherence to intestinal porcine epithelial cells (IPEC-1). One isolate, LR1, identified as Lactobacillus reuteri, was selected for extensive study of its probiotic and functional properties in IPEC-1 cell models. L. reuteri LR1 exhibited good adhesion to IPEC-1 cells and could inhibit the adhesion of enterotoxigenic Escherichia coli (ETEC) to IPEC-1 cells. L. reuteri LR1 could also modulate transcript and protein expression of cytokines involved in inflammation in IPEC-1 cells; the Lactobacillus strain inhibited the ETEC-induced expression of proinflammatory transcripts (IL-6 and TNF-α) and protein (IL-6), and increased the level of anti-inflammatory cytokine (IL-10). Measurement of the permeation of FD-4 showed that L. reuteri LR1 could maintain barrier integrity in monolayer IPEC-1 cells exposed to ETEC. Immunolocalization experiments showed L. reuteri LR1 could also prevent ETEC-induced tight junction ZO-1 disruption. Together, these results indicate that L. reuteri LR1 exhibits desirable probiotic properties and could be a potential probiotic for use in swine production.

• Food Engineering Progress
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• v.14 no.3
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• pp.208-216
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• 2010

Lab scale experiments were conducted in order to assess the applicability of $CaCO_{3}$-alginate beads as neutralizer for the high cell density cultivation and prepare the direct vat inoculation cultures of isolated sourdough lactic acid bacteria. With increasing the amount of bead and decreasing the diameter of bead in acidic solution, the neutralizing effect of $CaCO_{3}$-alginate bead became higher. In batch process with $CaCO_{3}$-alginate beads, Lactobacillus amylovorus DU-21 isolated from sourdough showed the highest viable cell counts and optical density in MRS broth. The values of viable cell counts and optical density were 9.996 log CFU/mL and 3.97, respectively. Experiments on the conditions which increase viability during lyophilization were carried out and the following results were obtained; 15% glycerol revealed the high cryoprotective effect on the concentrated cultures during lyophilization among the two cryoprotective agents. Consequently, $CaCO_{3}$-alginate beads and 15% glycerol were found to be useful not only to cultivate Lactobacillus amylovorus DU-21 but also to preserve strain.

• Food Engineering Progress
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• v.15 no.4
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• pp.346-354
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• 2011

This study was carried out to fortify the antimicrobial activity of yoghurt by adding liquorice extract to it. The liquorice extracts (1 mg/mL) showed relatively high antibacterial activity against H. pylori KCCM 40449 (p < 0.05). The solvent liquorice extracts of minimal inhibitory concentrations (MIC) against H. pylori KCCM 40449 were 25- 100 ${\mu}g$/mL. Lactobacillus amylovorus DU-21 with high EPS production ability were inoulated to milk after the addition of different amounts of liquorice extracts (0.0%, 0.05%, 0.1% and 0.2%). The physico-chemical characteristics of yoghurts added with liquorice extracts were examined. The initial pH, titratable acidity, viscosity and viable cell counts of the yoghurt added liquorice extracts were 3.41-3.51, 1.021-1.091%, 1,686-1,930 cp and 9.41-9.38 Log CFU/mL, respectively. The viscosity and syneresis of yoghurt were better than that of the control. Antimicrobial activity against H. pylori KCCM 40449 increased with increasing addition of liquorice extract. However, the sensory score of yoghurt added with different amounts of liquorice extracts was lower than that of the control (p < 0.05). As a result of the sensory evaluations, the flavor, taste, texture, color and overall acceptability of the yoghurt with 0.05% liquorice extract were found to be much better than those of the other groups (p < 0.05). Overall, the optimal amount of liquorice extract added in the manufacture of yoghurt was 0.05% of the total weight. Further studies on increment of antimicrobial activity and palatability of liquorice extract added yoghurt are necessary.

• Journal of Life Science
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• v.28 no.11
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• pp.1347-1353
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• 2018

The fermentation of non-digestible soy meal can convert polysaccharides into many compounds that have a wide variety of biological functions. Bacillus strains are capable of hydrolyzing non-digestible saccharides, such as melibiose, raffinose, and stachyose, found in soy meal components. A highly active ${\alpha}$-galactosidase (${\alpha}$-d-galactoside galactohydrolase, EC 3.2.1.22) was isolated from a bacterium in a traditional Korean fermented medicinal herb preparation. The isolate, T2-16, was identified as Bacillus coagulans based on its 16S rRNA sequence and biochemical properties, and the strain was named Bacillus coagulans NRR-1207. When incubated in 10%(w/v) skim milk, Bacillus coagulans NRR1207 caused a decrease in the pH of the culture medium, as well as an increase in titratable acidity and viable cell counts. This strain also showed higher activities of ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, naphthol-AS-BO-phosphohydrolase, and acid phosphatase when compared to other enzymes. It hydrolyzed oligomeric substrates, such as raffinose and stachyose, and liberated galactose, indicating that the Bacillus coagulans NRR1207 ${\alpha}$-galactosidase hydrolyzed the ${\alpha}$-1,6 glycoside linkage. These results suggest that the decreased stachyose and raffinose contents observed in fermented soy meal are due to this ${\alpha}$-galactosidase activity. Bacillus coagulans NRR1207 therefore has potential probiotic activity and could be utilized in feed manufacturing, as well as for hydrolyzing non-digestible soy meal components.

• Journal of Food Hygiene and Safety
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• v.33 no.6
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• pp.474-482
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• 2018

The aim of this study was to determine and analyze probiotic contents, pH, and acidity of 120 samples of health functional and processed foods containing lactic acid bacteria distributed in 2017. The changes due to the preservation methods were also determined and analyzed in five probiotic products. Two samples of the 85 health functional food products had lactic acid bacteria at lower levels than the marked amount required to meet Korean food standards, whereas the 35 processed products were all suitable for distribution. The averages for probiotic contents, pH, and acidity were $1.2{\times}10^{10}CFU/g$, 5.35, and 1.29%, respectively. The average count of lactic acid bacteria in the 17 samples with marked amount among the 35 processed foods was $5.8{\times}10^8CFU/g$. The effects of preservation temperature ($-20^{\circ}C$, $4^{\circ}C$, $20^{\circ}C$, and $40^{\circ}C$) and storage period (1, 3, and 6 months) on probiotic content, pH, and acidity were determined for 5 probiotic products. After 1 to 6 months, the average reduction in probiotic content was by 59%; the lowest reduction occurred at $4^{\circ}C$ and the highest reduction occurred at $40^{\circ}C$. In addition, 3 of the 5 products showed a rapid decrease in probiotic content by more than 70% at $40^{\circ}C$ after a storage period of 1 to 3 months. Therefore, from this study results, it is recommended that products containing lactic acid bacteria should be refrigerated and consumed shortly after purchase.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.384-397
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• 2018

The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.