Yun, Mid Eum;Lee, Ye Seul;Lee, Yun Ju;Park, Young Min;Park, Soo Nam
Applied Chemistry for Engineering
/
v.29
no.4
/
pp.452-460
/
2018
This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.
Kim, Min Ji;Cha, Min Kyeong;Lee, Do Kyung;Kang, Ju Yeon;Park, Jae Eun;Kim, Young Hee;Park, Il Ho;Shin, Hea Soon;Ha, Nam Joo
Korean Journal of Microbiology
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v.48
no.4
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pp.240-245
/
2012
Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes serious infection, particularly in immunocompromised patients. Also, P. aeruginosa possessing carbapenem-resistant metallo-${\beta}$-lactamases (MBL) has been reported with increasing frequency in Korea. We therefore analyzed the level of multidrug-resistant clinical P. aeruginosa isolated from a secondary hospital in Korea in 2010. A total of 92 isolates of P. aeruginosa were collected from Sahmyook Medical Center in 2010. Susceptibility to antimicrobial agents was determined by analysis of the minimum inhibitory concentration test; the inhibitor-potentiated disk diffusion (IPD) test was performed for MBL detection. RAPD-PCR was used for genotyping to rapidly characterize P. aeruginosa strains isolated from clinical patients. The percentages of non-susceptible isolates were as follows: 40.2% to ceftazidime, 58.7% to meropenem, 56.5% to gentamicin, 46.7% to tobramycin, 62.0% to ciprofloxacin and 97.8% to chloramphenicol. The 29 multidrug-resistant strains were screened by the IPD test: of the 21 PCR-positive isolates, 19 were IPM-1 producers and 2 were VIM-2 producers. Among the 19 IMP-1-producing P. aeruginosa isolates, 16 isolates showed similar patterns, and three different banding patterns were observed. The proportion of IMP-1-producing multidrug-resistant P. aeruginosa from clinical isolates steadily increased in this secondary hospital in Korea in 2010. This study provides information about the antimicrobial-resistant patterns and genotype of multidrug-resistant P. aeruginosa isolated from clinical isolates in Korea, 2010.
This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.3
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pp.320-326
/
2012
This study was conducted to investigate the antibacterial and antioxidative activities of water and ethanol extracts from a mixture of roasted coffee and red ginseng. The antibacterial effects of each extract were determined by the classical paper disc method. A water extract of mixture samples inhibited growth of all strains, but antibacterial effects were mostly weakened. Ethanol extracts showed stronger antibacterial effects than water extracts in all strains except Gram negative Escherichia coli and the fungi strain Candida albicans. Also, the antibacterial effect of the Bacillus cereus strain appeared in all samples, and the ES2 sample formed a clear zone of 19 and 20 mm against Pseudomonas aeruginosa and S. Typhimurium respectively (MIC=0.25 and 0.125 mg/mL). Determinations of free radical elimination for the different mixture extracts using 1,1-diphenyl-2-picrylhydrazyl (DPPH) were compared with ascorbic acid and butylated hyderoxytoluene as positive controls. The water and ethanol extracts of mixture samples (100 ${\mu}g/mL$) showed 55.38~60.01% and 59.37~70.50% DPPH scavenging activities, respectively. DPPH scavenging activities of all mixture samples were slightly higher than roasted coffee and red ginseng samples. However, DPPH scavenging activity decreased when red ginseng extract composed more than 70% of the total extract. The total polyphenol in the mixture samples measured by the Folin-Denis method revealed the highest level of polyphenol content in ethanol extract of sample 3, whereas polyphenol content differed with different mixture ratios, ranging from 105.16~119.79 mg/g in ethanol extract. In the water extract, the polyphenol content was greatest with water extract of sample 1, whereas in other samples the content varied from 93.75~109.18 mg/g.
In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.
This study was carried out to evaluate the antimicrobial effect of red ginseng (Panax ginseng C.A. Meyer) against several foodborne pathogens including Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus niger. The antimicrobial effect was determined by agar diffusion method using red ginseng extract, crude saponin and non-water-soluble fractions. Red ginseng extract showed antimicrobial effect against S. aureus, but not C. albicans or A. niger. The extract showed anti-bacterial activity at concentration above 30% against S. aureus, which cause both food poisoning and atophic dermatitis. Crude saponin showed antibacterial activity above 7.5% against the bacterium. However, the ginsenosides purified from crude saponin showed no antimicrobial activities at 100-200 ㎍/mL. To investigate the mode of growth inhibition, red ginseng extract and crude saponin were added to 0.85% NaCl solution containing S. aureus and then incubated at 35℃ for 12 h. The results showed that viable cells were rapidly reduced in above 10% concentration of red ginseng extract and above 2% of crude saponin, respectively. However, the crude saponin and red ginseng extract did not inhibit the bacterial cells completely at those same concentrations. On the other hand, whereas all non-water-soluble fractions showed inhibition zones above 10 mm against S. aureus, they showed no inhibition effects against E. coli, C. albicans or A. niger. The methanol fraction-1 (MF-1) showed the highest antibacterial activity against S. aureus, and the MIC (minimal inhibitory concentration) was 0.625 mg/mL. These results suggest that red ginseng extract, crude saponin and non-water-soluble fractions show selective antibacterial activity against S. aureus, and non-water-soluble fractions might be used as natural antibacterial agents.
This study was performed to evaluate the antioxidant and antimicrobial activity of methanol extract from Rosmarinus officinalis L. and its fractions. The ethyl acetate fraction of rosemary had a higher antioxidant activity in both DPPH ($3.22\;{\mu}g/ml$) and ABTS ($5.05\;{\mu}g/ml$) compared to other extracts and fractions. Based on the results of the FRAP assay, the ethyl acetate fraction of rosemary showed a value of $5.9{\pm}0.3\;{\mu}M/{\mu}g$, and buthanol fraction and rosmarinic acid exhibited values of $4.8{\pm}0.2\;{\mu}M/{\mu}g$ and $5.1{\pm}0.1\;{\mu}M/{\mu}M$, respectively. Measurements of the antimicrobial activities of the extracts, fraction against gram positive, negative bacteria revealed that the methanol extract, hexane, ethyl acetate, and chloroform fraction of rosemary caused Staphylococcus aureus and Escherichia coli to form clear zones greater than 12 mm. Furthermore, the methanol extract and chloroform fraction showed high antibacterial activity, with inhibition zone exceeding 13 mm. The methanol extract and chloroform fraction of rosemary had broad antimicrobial spectrums and low MIC values. Therefore, methanol extracts of rosemary could serve as potential antibacterial agents to inhibit pathogen growth in food and hand sanitizers.
Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
Journal of Life Science
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v.27
no.2
/
pp.202-210
/
2017
As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.
Kim, Mi-Hyung;Choi, Nam-Ki;Kim, Seon-Mi;Oh, Jung-Suk;Yang, Kyu-Ho
Journal of the korean academy of Pediatric Dentistry
/
v.32
no.2
/
pp.344-356
/
2005
There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from inhabitants of caries-free children's oral cavity, which inhibited the formation of artificial plaque by Streptococcus mutans and the production of volatile sulfur compounds by anaerobic bacteria. The isolates were identified by the test using API 50 CHL medium kit and 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4\;mg$, whereas being reduced to $5.2{\pm}2.0mg$ and $10.6{\pm}6.6mg$ in the media cultured with Streptococcus mutans and each isolate, respectively (p<0.05) 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8\;and\;2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively 5. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 6. When two isolates were tested with API 50 CHL medium kit, those were identified Lactobaciallius salivarius and Lactobaciallius delbrueckii subsp. lactis. 7. The similarity values of 16S rDNA sequence between each of isolates and Lactobaciallius salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobaciallius salivarius subsp. salicinius. These results indicated that two strains isolated from caries-free children's saliva, which inhibited the formation of artificial plaque and the production of volatile sulfur compounds, were identified as Lactobaciallius salivarius subsp. salicinius.
The purpose of this study was to investigate the distribution of microorganisms and the degree of contamination in the air of the dental clinics and to offer basic data as to the contamination of medical equipment and the prevention of the clinics. With this in mind, the researcher gathered air samples from the waiting rooms and medical offices of nine dental clinics in the city of J, South Korea with the use of a method of natural inattention and an air sampler and cultivated the samples on the plain table and drew from it bacteria falling and separated and sorted out the colony with the help of ATB and detected the distribution of the germs. The results are following, The number of bacteria falling in the air of the dental clinics was less than 10(CFU/Plate) with the exception of one dental clinic. This implies that they fit in with standards for hygiene. The number of bacteria falling in the air of the medical offices was less than 10(CFU/Plate) with the exception of one dental clinic. This implies that they fit in with standards for hygiene. The survey on the detection of staph. aureus reveals that all the dental clinics with the exception of B dental clinic proving to be positive had non-pathogenic staphylococci detected. The survey on the detection of pathogenic gram negative bacilli indicates that all the dental clinics but one were none detected. The survey on the distribution of germs shows that germs in 7 out of 9 dental clinics were none detected, and that they in four out of 9 waiting rooms were none detected. All the germs detected in the others were mostly non-pathogenic. The study shows that all the subject dental clinics but one were hygienically controlled and that there was a difference in accordance with cleaning and sterilization. This means that dental clinics should be equipped with systematic programs for cleaning and sterilization designed to prevent infection.
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