• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.157-162
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• 2009

An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

• Food Science and Preservation
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• v.18 no.1
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• pp.87-90
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• 2011

The methanol extracts of 19 commercial herb medicines was analyzed to antibacterial activities against Edwardsiella tarda, causing several fish diseases. Rhus javanica showed most strong antibacterial activity against E. tarda and Escherichia coli. Methanol extract of R. javanica was further extracted using several organic solvents having different polarity. Extract from ethyl acetate fraction showed strong activity against E. tarda as well as E. coli. Minimal inhibitory concentration, MIC of R. javanica extract was measured and resulted showing $64\;{\mu}g/m{\ell}$ for E. tarda and $256\;{\mu}g/m{\ell}$ for E. coli. It is needed that, from these results, further purification and isolation of reposible compound of these activities and further study on the synergy effect using combination with antibiotics against pathogenic bacteria.

• KSBB Journal
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• v.25 no.5
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• pp.429-436
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• 2010

Keratinase is widely used in certain industrial applications. The present study sought to improve the culture conditions of Bacillus subtilis SMMJ-2 to facilitate mass production of keratinase. Strain SMMJ-2 was irradiated by ultraviolet light and the resulting isolates were tested for keratinase activity. Isolates displaying elevated keratinase activity were selected and used to determine the optimum temperature (24, 30, 37, 45, $55^{\circ}C$) for bacterial keratinase production during a 4 day incubation period. The highest enzyme activity (55 units/mL/min), from a Bacillus subtilis SMMJ-2 mutant (mutant No. 2) was demonstrated following incubation at $30^{\circ}C$. The effects of carbon and nitrogen sources on keratinase production were confirmed by measuring the enzyme activity from the culture broth of the mutant strain cultured in various media containing different carbon source and nitrogen sources during a 4 day period. The optimal medium composition for producing keratinase consisted of 1% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, and 1.2% soybean meal. Optimal initial pH and temperature for producing keratinase were 7.0 and $30^{\circ}C$, respectively. Keratinases produced by B. subtilis SMMJ-2 and the mutant No. 2 were purified from the culture broth which used soybean meal as a nitrogen source. Membrane ultrafiltration, DEAE-sephacel ion exchange and Sephadex G-100 gel chromatography were used to purify the enzymes. The purified keratinases from both B. subtilis SMMJ-2 and the mutant No. 2 showed single bands and their molecular weights were estimated as 28 kDa and 42 kDa, respectively on SDS-polyacrylamide gel electrophoresis.

• Journal of the Korean Applied Science and Technology
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• v.34 no.4
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• pp.915-923
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• 2017

The by-products of red ginseng marc showing the improvement of health has been strongly studied because of the expectation of possibility to be used as the functional foods. This research was to investigate the extraction, separation, isolation, and evaluation of crude acidic polysaccharides related to antioxidant activity, sequently chemical structure of those products was evaluated by FT-IR and NMR. Compared with other solvents such as ethanol, methanol, propanol, acetone, and butanol, ethanol was selected. The concentration of red ginseng solution for extraction was selected as 10%(w/v) related to extraction yield(11.95%) and amount(11.8 mg/ml). In the ion exchange chromatography, aidic polysaccharides showing the highest antioxidant activity was obtained when eluted by distilled water. As results of structural analysis by FT-IR and NMR, peaks corresponding to C-O, C-O-O-, C-H, anomeric C-6, and repeated C-1 and C-6 linkages was to be presumed to be ($1{\rightarrow}6$) glycosidic linkage with the typical acidic polysaccharide.

• Journal of Korea Foresty Energy
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• v.20 no.2
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• pp.9-19
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• 2001

This study was carried out to investigate natural insectifuge materials from tree extractives in order to substitute for organic synthetic insecticides for food wrapping corrugated board. Tree samples were collected, extracted, fractionated with hexane, $CH_2Cl_2$, ethylacetate(EtOAc) and $H_2O$, and then freezed dried for further study. EtOAc or $H_2O$ fractions were chromatographed on a Sephadex LH-20 column for isolation and purification, and the isolated compounds were characterized by spectroscopic tools such as NMR and MS. Crude extractives of EtOAc and $H_2O$ fractions were added to the printing ink for corrugated board with the concentration of 2% or 3% based on the weight of the ink, then the prepared ink was printed on the corrugated board to be used for evasion test using larva of indian meal moth(Plodia interpunctella(Hubner)). Robtin, dihydrorobinetin and leucorobinetinidin were isolated from the wood extractives of black locust(Robinia pseudoacacia) and the bark of poplar(Populus alba $\times$ glandulosa) contained many kinds of compounds such as (+)-catechin, naringenin, aromadendrin, eriodictyol, sakuranetin and its glycoside, taxifolin, neosaturanin, salireposide, p-coumaric acid and aesculin. Much of (+)-catechin was isolated from the bark extractives of willow(Salix koreensis) in addition to (+)-gallocatechin and p-coumaric acid and the bark of weeping willow(Salix babylonica) also contained (+)-catechin, (+)-gallocatechin, dihydromyricetin and myricetin.

• Korean Journal of Medicinal Crop Science
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• v.11 no.2
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• pp.115-121
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• 2003

Essential oils from Fennel fruit(Foeniculum vulgare Mill), Olibanum resin(Boswellia carteii Birew) and Needl Juniperus stem(Juniperus rigida Sieb.) were extracted by a supercritical fluid extraction system(SFE) and biological activity of each essential oils were observed. SFE technique was applied for the isolation and purification of nonpolar biologically active essential oils from each samples. The quantitative analysis of essential oils was carried out by gas chromatography-mass spectrometer(GC/MS). About 60% of the growth of AGS and A549 cells were inhibited by adding 1.0g/l of the crude essential oils and below 40% was observed by the control. Cytotoxicity on human normal lung cell(HEL299) was scored as $15{\sim}18%$ for the crude essential oils and 12% for control, respectively. It meant that the essential oils were more effective than the control in anti-mutagenecity tested by CHO V79 cells. The effect of the essential oils on the growth of nerve cells, PC12 was observed as follows: The viable cell density was about two times higher than control.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.851-856
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• 2005

Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

• Food Science and Preservation
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• v.7 no.4
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• pp.409-413
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• 2000

Previously, the methanolic extracts of thirty Korean medicinal plant seeds were screened for tyrosinase inhibitors using a rapid and simple TLC method, which was superior to a conventional spectrophotometrical in vitro assay. As a result, the methanolic extracts of Thuja orientalis seeds was found to have strong tyrosinase inhibitory activity. To isolate active tyrosinase inhibitors, the seeds were defatted with n-hexane under reflux, and then extracted twice with methanol under reflux at 90$^{\circ}C$. The methanolic extract was evaporated to a small volumn in vacuo, and then successively fractionated with ether, ethyl acetate and n-butanol. The ether extract showing significant tyrosinase inhibitory activity was solubilized with 5% NaHCO$_3$and then acidified with 6N HCI. The ether souble acidic fraction was successively ohromatographed on silica gel, Sephadex LH-20 and preparative TLC. Among four compounds isolated, two of them showed stronger tyrosinase inhibitory activity, comparable to that of L-ascorbic acid (IC$\sub$50/=28$\mu\textrm{g}$/$m\ell$). These results suggest that Thuja orientalis seeds may be useful as potential sources of antibrowning agents in fruits and vegetables, and anti-melanoma agents in cosmetics and phamaceuticals.

• Korean Journal of Organic Agriculture
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• v.29 no.1
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• pp.97-109
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• 2021

Plant parasitic nematodes are causing significant damage in crop production. There is a need to develop eco-friendly nematicide that reduces the damage of nematode and has little effect on the environment and human. In this study, we have isolated a substance having nematicidal activity from Ginkgo biloba L. outer seedcoat. Studies of G. biloba L. outer seedcoat are insufficient compared to the seed and leaves due to their odor and toxicity. The dried G. biloba L. outer seedcoat was extracted with dichloromethane:methanol (1:1) and fractionated into hexane, ethyl acetate and H₂O. Four steps TLC were performed from EtOAc fraction to purely isolate GB4-3 with nematicidal activity. To compare nematicidal activity, G. biloba L. seedcoat methanol extract and purified GB4-3 were investigated in terms of treatment concentration and time. As a result, the nematicidal activity increased with concentration and time. In the place treated with 20 ㎍/mL of crude G. biloba L. seedcoat MeOH extract, strong activity appeared after 12 hours, and 46% nematicidal activity shown after 18 hours. About 69% of nematicidal activity was confirmed in the place where GB4-3 purified from outer seedcoat was treated with 20 ㎍/mL, and the possibility of development as nematicide was very high. This study could be used as a basic data for the development of a nematode preparation from G. biloba L. outer seedcoat.

• Applied Biological Chemistry
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• v.20 no.2
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• pp.228-235
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• 1977

The yeast strain was isolated from food industry waste water and its identification and biological characteristics were investigated. The optimum condition for cultivations and its activities for the reduction of B.O.D. on the food industry waste water were also confirmed. The results are as follows; 1) The isolated was identified as Candida curvata. 2) Candida curvata grew well in all of the experimented media, so and it can be regarded as a useful strain in the treatment of food industry waste water. 3) There was only a slight difference in the induction period between sterilized cultivation and unsterilized cultivation. But in the ice cream waste water, the period was considerably longer in unsterilized cultivation. 4) Specific rate of growth of Candida curvata in sugar waste water was 0.50/hr, ice cream waste water 0.50/hr, and beer waste water 1.0/hr. 5) Increasing of innoculum reduced the induction period in unsterilized cultivation. 6) The amount of dried yeast from sugar waste water were $175mg/{\ell}$, ice cream waste water $628mg/{\ell}$, and beer waste water $857mg/{\ell}$. Crude protein content in the dried yeast from sugar waste water were 52%, ice cream waste water 54%, and beer waste water 54%. 7) The rate of BOD reduction in sugar waste water were 49%, ice cream waste water 80%, and beer waste water 64%.