• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.168-174
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• 2011

In order to develop safe and economic novel antifungal agents, we prepared 73 methanol extracts from medicinal and edible herbs and examined their 365 solvent fractions using n-hexane, methylene chloride, ethylacetate, butanol and water residue based on the sequential organic solvent fraction method. When using the various fractions in the screening step for antifungal activity, we discovered ethylacetate fraction of Morus alba L., methylene chloride fraction of Pimpinella brachycarpa (MCPB), and n-hexane fraction of Salvia miltiorrhiza Bunge, which all have activities in methanol extracts, as potential sources of antifungal agents. Amongst these, the antifungal activity of P. brachycarpa has not to date been reported on. In addition, the mycelial growth inhibition and spore germination inhibition activities of MCPB against A. niger were confirmed by disc-diffusion assay in a 10 day culture. The MIC and MFC of MCPB were determined as 0.25 and 0.5 mg/ml, respectively. The MCPB has no hemolytic activity against human RBC at 0.5 mg/ml and glycoside-flavonoids are theorized to be active constituents. These results suggest that MCPB has a prominent antifungal activity and that the application of sequential organic solvent fractions, instead of simple natural product extracts, is useful in the screening process of novel bioactive substances.

• Journal of Life Science
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• v.23 no.3
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• pp.361-367
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• 2013

Hizikia fusiformis has been widely used in Oriental herbal medicine and health food. To identify antioxidation properties that contain natural bioactive substances, we investigated the distribution of active compounds existing in batches of organic solvent fractionation. A dried form of H. fusiformis was subjected to sequential fractionation using n-hexane, ethyl acetate, n-BuOH, and aqueous n-BuOH. The results showed that among the four isolated fractions, the n-BuOH fraction showed the highest antioxidation activities. The n-BuOH fraction was applied to reserve-phase silica gel column chromatography, which produced three fractions: BA, BB, and BC. Among these fractions, BB showed the highest antioxidation activities, which increased in a concentration-dependent manner. At a concentration of 1.0 mg/ml n-BuOH fraction, the activities of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and reducing power were approximately $45{\pm}0.14%$ and $1.34{\pm}0.23$, respectively. In addition, the activities of ${\beta}$-carotene-linoleic acid, hydrogen peroxide scavenging, and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) radical scavenging were $76{\pm}0.12%$, $82{\pm}0.06%$, and $65{\pm}0.17%$, respectively. These findings suggest that the BB fraction contains potent antioxidation properties and that it could be used in the production of natural and functional foods.

• Korean Journal of Weed Science
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• v.14 no.1
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• pp.42-48
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• 1994

The toxic compounds of sorghum stem extracts were isolated by different organic solvents and pH, and characterized and quantified in terms of their inhibition of seed germination and seedling growth in Echinochloa colona(L.) Link and radish(Raphanus sativus L.). Sequential partitioning of stem extract with various organic solvents with increasing polarity showed that all fractions of hexane, ethyl ether, methylene chloride, ethyl acetate, and the aqueous remainder inhibited germination and seedling growth in E. colona. Of the five fractions, the ethyl ether fraction had the greatest inhibitory effect on E. colona. Further separation of the ethyl ether fraction at different pH(pH 2-11) showed that phytotoxic compounds were acidic. The result indicates that the phytotoxin present in the stem extract may be nonpolar and acidic.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.371-376
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• 2009

The fruits of sweet cherry are highly appreciated by the consumer due to their precocity and quality, such as their sweetness, color and sourness. In this study, the hot-water extract and its sequential organic solvent fractions were prepared from domestic Napoleon sweet cherry (Prunus avium L.) to investigate antimicrobial and antioxidant activity. The hot-water extract contained about 40% sugars, and the solvent fraction yields for hexane, ethylacetate (EA), butanol, and water residue were 0.01%, 3.45%, 16.30%, and 80.24%, respectively. Contents of total polyphenol and total flavonoid of the fractions were 1.24~5.24%, and 0~3.76%, respectively. Among the fractions, EA fraction showed the highest total polyphenol and total flavonoid concentrations. Evaluation of antimicrobial activity of the extract and fractions revealed that EA fraction and butanol fraction contained strong antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Salmonella typhimurium with minimal inhibitory concentration (MIC) of 0.5~1.0 mg/mL. But the extract and fractions tested were not active to Pseudomonas aeruginosa. In a while, only hexane fraction showed anti-Candida activity with 0.5~1.0 mg/mL of MIC. The fraction showed strong activity against different multi-antibiotics resistant strains such as C. albicans CCARM 14020. Antioxidative activity assay showed that EA fraction has a strong DPPH scavenging activity and a reducing power. The $IC_{50}s$ of vitamin E and EA fraction were 15.5 and $195.5\;{\mu}g/mL$, respectively. Our results suggest that the fruit of P. avium L. has high potentials with anti-Candida and antioxidative activity.

• Journal of Life Science
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• v.25 no.7
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• pp.757-764
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• 2015

The immature fruit of Rubus coreanus Miquel (bokbunja in Korean) is mainly consumed as a fruit wine due to its sour taste and low sugar content. The lees (LBW) remaining after the production of bokbunja wine are discarded as they have no specific usage. The aim of this study was to develop high-value-added biomaterials for functional foods and beauty/health products by investigating the anti-microbial, anti-oxidant, and anti-thrombosis activities of LBW using ethanol and hot water extracts and their subsequent organic solvent fractions. The ethyl acetate (EA) fraction of LBW extracts has a high polyphenol content (413–459 mg/g), and showed strong anti-microbial activity against gram-positive bacteria. The EA fraction also showed excellent radical-scavenging activity against DPPH anion, ABTS cation, and nitrite, with strong reducing power. The polyphenol-enriched EA fraction strongly inhibited thrombin, prothrombin, and blood coagulation factors. The butanol fraction showed a specific inhibition of coagulation factors, as measured in activated partial thromboplastin time assay, which is linked to intrinsic blood coagulation. The butanol fraction also showed strong inhibition of platelet aggregation, at levels comparable to aspirin. The residue of the hot-water extract, which is produced by sequential solvent fractionation of the LBW extract, showed superior inhibition against platelet aggregation when compared to aspirin. Our results suggest that the LBW, which are currently discarded, are a promising source of novel functional foods and beauty/health products.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.16-20
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• 2000

The aqueous extract from the cultural mycelium of Paecilomyces japonica showed cytotoxicity against several tumor cells including Jurkat, U937, HL-60, HepG2, BW5147.G.1.4, and NIH3T3. When the aqueous extract was fractionated by sequential organic solvent extractions using n-hexane and ethyl acetate, the ethyl acetate fraction appeared to contain the most cytotoxic activity, and the $IC_{50}$ values for various tumor cells were in the range from 1.5 to $10.0{\;}\mu\textrm{g}/ml$. To elucidate the cellular mechanism underlying the induced cytotoxicity, the apoptotic DNA fragmentation along with the cell cycle proression was examined in Jurkat T cells following the ethyl acetate fraction treatment. In the presence of $2.5{\;}\mu\textrm{g}/ml$ of the ethyl acetate fraction, apoptotic DNA fragmentation of the cells was detected within 1 h and increased upto 24 h in a time-dependent manner. Under the same conditions, a sub-G1 peak was detectable by flow cytometry. These results indicate that the cytotoxic effect of P. japonica on tumor cells is attributable to the induced apoptosis.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.144-151
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• 2012

In the course of this study aimed at the development of functional food ingredients from seaweeds, the in vitro antimicrobial activities of methanol extracts prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta) were determined against food-borne diseases and pathogenic microorganisms including multi-drug resistant (MDR) Pseudomonas sp. and Candida sp. Based on disc-diffusion assays at 500 g/disc concentration of the methanol extracts, Ishige okamurai, I. foliacea, Sargassum confusum, and S. yamade exhibited strong antibacterial activities in a broad-spectrum, except against Pseudomonas aeruginosa. In addition to the latter four seaweeds, Ecklonia stolonifera, E. cava and Eisenia bicyclis also demonstrated antifungal activity against C. albicans. Among these 8 selected seaweeds, I. okamurai, I. foliacea, and S. yamade exhibited strong hemolytic activity (55-93%) at 500 g/ml against human RBC. Organic solvent sequential fractions using hexane, ethylacetate and butanol, and water residues were prepared from the 8 selected seaweeds and their anti-Candida sp. activities were further determined. The ethylacetate and butanol fraction of I. okamurai, and the hexane fraction of I. foliacea demonstrated antifungal activity against MDR-pathogenic Candida sp. Although the solvent fractions had no activity against MDR-Pseudomonas sp., our results suggest that seaweeds, especially Ishige okamurai, I. foliacea, S. confusum, and S. yamade could be developed as broad-spectrum antimicrobial ingredients.

• Microbiology and Biotechnology Letters
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• v.38 no.1
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• pp.77-83
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• 2010

The fructus of Crataegus pinnatifida Bunge (CBF) has been used as medicinal and food source in worldwide. In this study, antimicrobial activity of the methanol extract and its sequential organic solvent fractions of CBF against different pathogenic bacteria and fungi, including multidrug resistant Pseudomonas aeruginosa and Candida sp., were investigated. The methanol extract of CBF was active against various gram-positive and gram-negative bacteria, and the ethylacetate and butanol fractions of CBF showed strong antibacterial activity against Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Salmonella typhimurium, Proteus vulgaris, Escherichia coli and various multidrug resistant Pseudomonas aeruginosa with minimal inhibitory concentration of 1.0~7.5 mg/mL. Also the fractions showed anti-Candida activity against C. albicans, C. kruseis and C. geochares. The methanol extract of CBF and its solvent fractions, except n-hexane fraction, did not show any hemolytic activity against human red blood cell up to $500\;{\mu}g/mL$, respectively. The hemolysis in n-hexane fraction at $500\;{\mu}g/mL$ was less than 9.9%. Our results suggest that the CBF could be developed as a potent antibacterial agent, especially for multidrug resistant Pseudomonas aeruginosa.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.3
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• pp.303-309
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• 2014

The fruit of Gleditsia sinensis has been extensively used as a key ingredient of an herbal remedy for the treatment of various inflammatory diseases in traditional Korean Medicine. However, the reason of using the fruit of G. sinensis for the remedy is unclear. Since Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a key anti-inflammatory transcription factor, which is activated by the fruit of G. sinesis, we examined whether other plant parts of G. sinensis are also capable of suppressing inflammatory responses by activating Nrf2. Water extracts of various parts of G. sinensis were prepared and tested for Nrf2 activation by reporter assay and western blot analysis. Our results show that the hull of G. sinensis is the most potent in activating Nrf2. Sequential organic solvent extraction of the hull show that all the fractions had a higher potency in activating Nrf2 than the water extract, albeit differential degrees. The hull originated from Korea in general activated Nrf2 strongly compared to that of China. Chloroform fraction of the hull was further examined, showing that the fraction induced nuclear localization of Nrf2, indicative of activated Nrf2, and Nrf2-dependent gene expression including NAD(P)H dehydrogenase quinone 1 (NQO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and heme oxygenase - 1 (HO-1). Therefore, our results show that, among other plant parts examined in this study, the hull of G. sinensis is the most potent, providing the experimental basis for the use of the hull of G. sinensis as an active ingredient for an anti-inflammatory remedy.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.195-200
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• 2008

Saxifraga stolonifera (Saxifragaceae) is a perennial herbaceous plant growing in Korea, China, Japan and Russia. The aerial part has been used as herbal medicine for treatment of pneumonia, frostbite, inflammation and microbial infection. In this study, fresh juice and methanol extract were prepared from the aerial part of S. stolonifera, and their antimicrobial, antithrombin, and antioxidant activity were evaluated, respectively. The fresh juice showed weak antimicrobial activity against Proteus vulgaris, Escherichia coli O157:H7 and Candida albicans with ignorable DPPH scavenging activity. But, the methanol extract showed strong antioxidant activity ($IC_{50}$ of $37.5{\mu}g/mL$) with minor, broad-range antimicrobial activity. Antithrombin activities were not observed in fresh juice and extract, up to 1.5 mg/mL. Sequential organic solvent fractionation of methanol extract showed that $IC_{50}s$ of ethylacetate and the butanol fraction were 6.9 and $7.8{\mu}g/mL$, respectively, that is comparable with vitamin C or butylated hydroxytoluene. Analysis of component in extract and fractionates suggested that the antioxidants in fractions are diverse and the active substances have glycosylated phenolic structure. Our results suggest that the aerial part of S. stolonifera could be used as the natural source of potential antioxidant.