• The Korea Journal of Herbology
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• v.32 no.2
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• pp.87-95
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• 2017

Objectives : Infectious diseases by Methicillin-Resistant Staphylococcus aureus (MRSA) are a growing problem worldwide. Characteristic of MRSA is endlessly mutation to resist antibiotics. Daehwanggo (DHG) is one of the oriental medicine prescriptions contained in Principles and Practice of Eastern Medicine. Daehwanggo was mainly used for external preparation from old times. The purpose of this study is to confirm possibility as supplementary drug of DHG about antibiotics through observation of synergy effect between DHG and commercial antibiotics and to observe restriction on growth of MRSA on any pathway through observation of mechanism. Methods : The minimum inhibitory concentration (MIC) of DHG against MRSA is $500{\sim}2000{\mu}g/m{\ell}$ by broth dilution method. In the checkerboard method, the combinations of DHG with antibiotics has partial synergistic effect or synergy effect and DHG markedly reduced the MICs of the antibiotics oxacillin (OX), gentamicin (GT) against MRSA. In the inhibition of resistance mechanism of DHG against MRSA, the expression of resistance gene and protein about ${\beta}-lactam$ antibiotic was reduced. Also, we observed the effect of DHG about cell membrane permeability against MRSA, and confirmed that DHG suppressed growth of strains by increasing cell membrane permeability. Results : Basis on the result, we speculate that DHG increase antibacterial activity of antibiotics against MRSA by changing the structure of cell wall of MRSA. Conclusions : These data suggest that Daehwanggo possesses possibility as supplementary drug about antibiotics against MRSA.

• 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 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.

• Journal of the Korean Applied Science and Technology
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• v.38 no.6
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• pp.1383-1392
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• 2021

It has been reported that emodin, a major pharmacologically active ingredient of herbal medicines such as Polygonum cuspidatum, Polygonum multiflorum, Rheum palmatum, and Aloe vera, is effective in antioxidant, antibacterial, anti-inflammatory, anticancer, and liver protection. In this study, to investigate the potential of emodin to be used as a skin disease and functional material, the activity related to the improvement of inflammation and skin barrier function was confirmed. To observe the anti-inflammatory effect on HaCaT cells, which are human keratinocytes, cytokine inhibition was confirmed by ELISA kit and protein expression by western blot. In HaCaT cells activated with TNF-α (10 ng/mL)/IFN-γ (10 ng/mL), emodin was treated with each concentration (5, 10, 20, 40) µM. As a result, It was confirmed that the production amount of TNF-α, IL-1β and IL-6 decreased as the concentration of emodin increased. In the experimental results on the expression levels of inflammation-related proteins iNOS and COX-2, it was confirmed that 48% of iNOS and 29% of COX-2 were inhibited compared to control at a concentration of 20 µM of emodin. As an indicator of skin barrier function improvement, the mRNA expression level of filaggrin, involucrin, and loricirn and the production amount of filaggrin, involucrin, and loricirn were confirmed. and excellent results were obtained with an emodin concentration-dependent increase. In particular, filaggrin, which was produced twice as much as the control at a concentration of 20 µM, is a protein involved in the formation of NMF, a natural moisturizing factor, and is known to play an important role in moisturizing the stratum corneum. In conclusion, it was confirmed that emodin can be used as a material for improving inflammation and improving skin barrier function, which is part of the potential for use as a skin disease and functional material. It is believed that if additional research is performed in the future, the scope of its application can be further expanded.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.52-67
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• 1998

The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.237-245
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• 2004

The physicochemical and physiological activities of domestic garlic from 3 different areas (Namhae, Jeju and Uiseong) were analyzed. The contents of moisture, ash, crude protein and crude fiber in garlic were little different in 3 kinds of area. Total sugar and water soluble phenolic compounds were higher in garlic from Namhae. The free sugars found in garlic were fructose, sucrose and lactose. Five kinds of organic acids were determined. Malonic acid and citric acid contents in garlic from Namhae were 23.7${\pm}$1.16 mg% and 22.1${\pm}$0.82 mg%, respectively. Total mineral content of garlic samples were in a range of 7112.6～9067.3 mg%, the potassium content showed the highest concentration (4117.3${\pm}$7.19～5175.3${\pm}$9.61 mg%). The electron donating abilities in 0.2% of garlic from Namhae and Uiseong showed 46.2${\pm}$1.25% and 37.0${\pm}$1.l6%, respectively. The nitrite scavenging effect was measured at different conditions (pH 1.2 and 4.2). The nitrite scavenging effects were higher at pH 1.2, and reached more than 95% by adding 0.2% and 0.1 % of garlic juice at pH 1.2. Addition 0.02～0.001 % garlic juice in showed the SOD-like activities. Its activity of garlic from Namhae. was a range of 6.0${\pm}$0.37～14.4～0.69%. It was found that 0.2% and 0.1% garlic showed strong antimicrobial action against growth of all the tested bacteria. Antimicrobial action. was showed 74.7${\pm}$0.70% and 51.7${\pm}$1.l1% on Sal. typhimurium in 0.2% of garlic from Namhae and Uiseong and 28.6${\pm}$0.90% on B. subtilis in garlic from Jeju.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.3
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• pp.386-395
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• 2016

To develop a functional fermentation food from Rubus coreanus Miquel (Bokbunja) and chlorella mixtures, optimal lactic acid fermentation conditions were established, and quality properties based on physicochemical evaluation such as chemical compositions, free sugars, organic acids, and antibacterial activities were investigated. Regarding optimal fermentation strain selection, formation of lactic acid was best in Lactobacillus plantarum among the experimental strains (10 kinds), and the optimal fermentation temperature was $37^{\circ}C$. In addition, overall acceptability in the sensory evaluation was highest in the 5% chlorella mixture sample. Therefore, quality properties of the prepared sample under the established optimal fermentation conditions were investigated. Moisture, total sugar (dry basis), crude fiber (dry basis), and pH of fermented Rubus coreanus Miquel juice (RCM) with 5% chlorella mixture (RCM-C5) were reduced by 4.90%, 14.15%, and 0.32%, respectively, as compared with non-fermented RCM. Meanwhile, crude protein, crude fat, and crude ash (dry basis) of RCM-C5 were elevated by 13.75%, 0.18%, and 0.73%, respectively, as compared with RCM. The yellowness (b value) of color values was greater in RCM-C5 compared to RCM. The free sugar and organic acid contents of RCM-C5 were elevated by 0.97% and 616.30 mg%, respectively, as compared with RCM. In addition, the gram positive bacterium Staphylococcus aureus was elevated by 5.83% while gram negative bacteria Escherichia coli and Salmonella Typhimurium were elevated by 2.94% and 4.67%, respectively, as compared with RCM. In conclusion, the quality properties of RCM and chlorella lactic acid fermentation mixtures were improved compared with the general RCM product. Consequently, it is possible to apply fermented RCM as a functional fermentation food.

• Journal of agriculture & life science
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• v.46 no.4
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• pp.129-139
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• 2012

This study was carried out to evaluate the chemical properties and functional characteristics, such as general composition and bioactivity compounds contents of fresh and blanched (at $95^{\circ}C$, for 5min) garlic shoot from Namhae. Also, evaluated antioxidant and antimicrobial activities of water and ethanol extract of fresh and blanched garlic shoot. The moisture content of fresh garlic steam was $ 85.14{\pm}0.35%$, crude protein and crude lipid were $0.79{\pm}0.26%$ and $2.96{\pm}0.03%$ respectively. Vitamin C content was higher in fresh garlic shoot ($7.07{\pm}0.84mg/100g$) than blanched. Total phenol and total chlorophyll contents were respectively $16.93{\pm}1.17mM/g$ and $6.70{\pm}0.46mg/g$ in fresh garlic shoot. Allicin content of blanched garlic shoot was $128.63{\pm}1.59mM/g$. This content was 1.82 times higher than the fresh garlic shoot. Total pyruvate content was higher in fresh garlic shoot ($24.63{\pm}1.59mM/g$), but thiosulfinate was higher in fresh garlic shoot. Total flavonoide was the highest in water extract of blanched garlic shoot ($3.67{\pm}0.00mM/g$). ABTS radical scavenging activity of water extract form blanched garlic shoot was $85.09{\pm}0.28%$, which was higher than the other extracts. NO radical scavenging activity of ethanol extract from blanched garlic shoot was significantly higher than the extracts from fresh garlic shoot. Antibacterial activity to S. aureus, S. enterica, B. cereus and E. coli was only indicated in water extract of fresh garlic shoot.