• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1706-1712
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• 2006

Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1672-1678
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• 2009

We studied the effects of aged total petroleum hydrocarbons (aged TPH) on the survival of allochthonous diesel-degrading Rhodococcus sp. strain YS-7 in both laboratory and field investigations. The aged TPH extracted from a crude-oil-contaminated site were fractionized by thin-layer chromatography/flame ionization detection (TLC/FID). The three fractions identified were saturated aliphatic (SA), aromatic hydrocarbon (AH), and asphaltene-resin (AR). The ratio and composition of the separated fractions in the aged TPH were quite different from the crude-oil fractions. In the aged TPH, the SA and AH fractions were reduced and the AR fraction was dramatically increased compared with crude oil. The SA and AH fractions (2 mg/l each) of the aged TPH inhibited the growth of strain YS-7. Unexpectedly, the AR fraction had no effect on the survival of strain YS-7. However, crude oil (1,000 mg/l) did not inhibit the growth of strain YS-7. When strain YS-7 was inoculated into an aged crude-oil-contaminated field and its presence was monitored by fluorescent in situ hybridization (FISH), we discovered that it had disappeared on 36 days after the inoculation. For the first time, this study has demonstrated that the SA and AH fractions in aged TPH are more toxic to an allochthonous diesel-degrading strain than the AR fraction.

• 대한본초학회지
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• 제31권3호
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• pp.71-75
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• 2016

Objectives : Mori Ramulus is a young branch of Morus alba Linne, which has been used as traditional medicines for antidiarrhea, antibacteria, and antihypertension in Korea. With the basis on these medicinal activities, it is registered at the Korean Herbal Pharmacopoeia. The standard for quality control in use, however, is not yet validated.Methods : We experiment on heavy metals test, TLC, loss on drying, ash, acid-insoluble ash, contents of ethanolsoluble extracts and contents of oxyresveratrol by using the 15 samples of Mori Ramulus which are collected domestically. We tested items according to the General Test, processes and apparatus, Crude Drugs Test of KP(2011).Results : Purity test(Heavy metals) showed contents for Pb, As, Hg, and Cd range of 0.1~1.1 ppm, 0.0~0.2 ppm, 0.00~0.01 ppm, and 0.00~0.02 ppm. Identification test(Thin layer chromatography) was on comparing with morucine standard solution in R_f value, all samples showed morucine spot (dark blue, R_f : 0.3) as same as R_f value of morucine-standard solution. Loss on drying was ranged from 5.1 to 8.9 %, ash and acid-insoluble ash were between the range 3.0~6.3% and 0.3~0.8%. Contents of ethanol-soluble extracts was ranged from 6.2 to 12.0%. Content of oxyresveratrol was ranged from 0.01 to 0.16 % based on the oxyresveratrol standard curve.Conclusions : We sincerely hope that this study will be contributed to the standardization and quality control of Korean herbal medicines.

• Journal of Periodontal and Implant Science
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• 제23권2호
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• pp.243-259
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• 1993

The bone resorbing activity of $PGE_2$ and elevated level of prostaglandins(PGs) and thromboxanes (TXs) in inflamed gingiva which are cyclooxygenase(C) metabolites have been well documented. Nonsteroidal anti-inflammatory drugs(NSAIDs) have been known to suppress gingival inflammation and bone resorption through the specific inhibitory action on the C pathway thereby decrease of various C metabolites. Recent studies provide unequivocal results that gingival tissue metabolizes arachidonic acid(AA) mainly through lipoxygenase(L) pathway. And the results of our previous experiments suggest that indomethacin may have inhibitory action on L as well as C. Thus we started this study to show the influences of several C inhibitors on the L activity at therapeutic and toxic dosage. Periodontal tissue samples were obtained from patients with advanced periodontitis and incubated with $^{14}C-AA(0.2{\mu}Ci)$ and various enzyme inhibitors. The tissue lipid extracts were separated by means of thin layer chromatography(TLC) and analyzed by means of autoradiography and TLC analyzer. Our results showed that aspirin inhibited C more selectively than L, however at higher concentration it also decreased HETEs production significantly. Indomethacin showed dose-dependent inhibition of L as well as C and all of the L metabolites were decreased to the same degree by high concentration of indomethacin. AA-861, which is an experimental tool of selective L inhibitor, showed inhibition of HETEs production but no effect on the production of $TXB_2$, PGs and $LTB_4$. Various propionic acid derivatives NSAIDs(ibuprofen, flurbiprofen, naproxen) showed the same patterns of effect on AA metabolism each other that was profound inhibition of PGs production, to the less degree HETEs and $TXB_2$ production, and of no effect on the $LTB_4$ production.

• 한국균학회지
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• 제14권3호
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• pp.201-207
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• 1986

효모 세포(S. uvarum)를 재료로 하여 배양 시기 및 sugar starvation시기에 특이하게 출현 또는 유도되는 RNase의 localization과 특성을 조사하고자 하였다. 정상 배양 시기 및 sugar starvation시킨 효모 세포를 세포 분획구에 따라 RNase 활성도를 측정하는 한편 ribosome 양의 변화를 조사하였다. 특히 ribosomal 분획구에서 추출한 RNase들을 poly(C)와 반응시킨 후 생성물을 TLC에 적응하여 효소의 특성 및 유도 여부를 조사하였다. 그 결과를 요약하면 다음과 같다. 세포 분획구 중 $45,000{\times}g$ pellet 분획구 및 Postribomosal 분획구에서는 배양 시기나 sugar starvation에 관계없이 RNase의 활성도는 유의하게 증감하지 않았으나, ribosomal 분회구에서는 정체기와 sugar starvation시 활성도가 각각 2배, 10배 이상 급격히 증가하였다. ribosome의 양적 동태를 살펴보면 early log phase의 세포에 비하여, 정체기 세포와 sugar starvation시킨 세포에서는 $1/3{\sim}1/6$까지 급격히 감소하였다. TLC의 결과 rRNase의 종류는 early log phase에서는 oligonuclease와 3'-ribonuclease, 5'-ribonuclease,stationarf phase에서는 oligonuclease, 3'-ribonuclease, sugar starvation 시켰을 때는 3'-ribonuclease, 5'-ribonuclease의 활성이 나타났다. 그리고 완전 배지를 사용한 효모 세포에서는 공통적으로 oligonuclease의 활성이 나타난 반면, sugar starvation시킨 효모 세포에서는 oligonuclease의 활성은 나타나지 않았다.

• 한국미생물·생명공학회지
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• 제33권2호
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• pp.112-116
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• 2005

스웨덴 산림지역의 토양시료로부터 분리한 Bacillus licheniformis KMU-3의 작물병원성 곰팡이에 대한 항진균 활성 및 항진균물질의 생산조건에 대하여 검토했다. B. licheniformis KMU-3은 Rhizoctonia solani KACC 40101에 대하여 높은 항진균활성을 나타냈었다. 항진균물질 생산을 위한 최적배양조건은 탄소원으로 $1.0\%$ sodium acetate, 질소원으로는 $1.0\%$ ammonium sulfate, $24^{\circ}C$, pH 8.0, 180 rpm, 24시간 진탕배양하였을 때 가장 높은 항진균활성를 나타내었다. TLC상에서 Rf값이 0.49인 밴드에 나타난 물질에서 항진균활성이 확인되었으며, 이물질은 Rhizoctonia solani AG-4 KACC 40142, Colletotrichum orbiculare KACC 40808, Colletotrichum gloeosporioides KACC 40804, Cladosporium cucumerinum KACC 40576, Fusarium graminearum KACC 41040, Fusarium oxysporum KACC 40052에 대하여서도 높은 항진균활성을 나타내었다.

• Restorative Dentistry and Endodontics
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• 제15권1호
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• pp.153-164
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• 1990

Human dental pulps obtained from normal teeth, hypersensitive teeth and teeth with inflamed pulp were studied to measure and to compare the endogenous levels of arachidonic acid metabolites in order to see the relative activities of the different pathways involved in arachidonic acid metabolism. Pulp homogenates were incubated with $^{14}C$-arachidonic acid and lipid solvent extracts were separated by thin layer chromatography (TLC) to be analyzed by autoradiography and TLC analyzer. 1. The most significant metabolite was HETEs showing $96.9{\pm}37.8$pmol/mg tissue protein/hr in normal pulp, $169.2{\pm}76.7$ in hypersensitive pulp and $385.4{\pm}113.2$ in inflamed pulp. In normal pulp $LTB_4$, 6-keto-$PGF_{1\alpha}+PGE_2$, $TXB_2$ and unidentified metabolite were formed in decreasing order. While in hypersensitive and inflamed pulp 6-keto-$PGF_{1\alpha}+PGE_2$, $LTB_4$, $TXB_2$ and unidentified metabolite were formed in decreasing order. 2. In hypersensitive pulp only HETEs were significantly increased when compared with that in normal pulp. The levels of all the converted metabolites in inflamed pulp were significantly increased compared with those in normal pulp. In inflamed pulp, the levels of $TXB_2$ and HETEs were significantly increased compared with those in hypersensitive pulp. 3. The ratio of each metabolites to the total converted metabolites showed an increased value of $TXB_2$ and 6-keto-$PGF_{1\alpha}+PGE_2$, as the degree of inflammation was increased, while that of HETEs decreased both in hypersensitive pulp and inflamed pulp more than in normal pulp. 4. The relative amounts of the total metabolites formed in lipoxygenase pathway to cyclo-oxygenase pathway were 6.8 fold in normal pulp, 4.4 fold in hypersensitive pulp and 3.8 fold in inflamed pulp.

• 한국식품영양과학회지
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• 제23권2호
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• pp.251-260
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• 1994

Lipophilic brown pigments produced during the fermentation of domestic Meju and Doenjang were fractionated by column and thin -layer chromatography (TLC). Each of the fractions was tested for the antioxidant activity and then characterized by spectroscopic analysis. The lipophilic brown pigments were separated into chloroform -soluble and methanol-soluble parts in which Meju resulted the higher content of chloroform-soluble part than that of methanol-soluble part ; however, Doenjang exhibited the opposite result to that of Meju. More strong antioxidant activity was found in the methanol-soluble part than the chloroform-soluble part. Four and five fractions were separated from chloroform-soluble and methanol-soluble parts respectively. by silicagel TLC. The fraction that exhibited the high antioxidant activity showed a strong absorption at 260nm caused by amino compounds in UV spectrum The other fractions which did not have antioxidant activity absorbed at 240nm by carbonic acid and it ester,. IR spectrum of each fraction commonly showed absorption at 3400cm-1 , 2800cm-1 , 1700cm-1, 1600cm-1, 1400cm-1 , 1300cm-1 and 1100cm-1. Especially , the fraction which had a strong antioxidant activity showed absorption at 2800cm-1, 1400cm-1, 1600cm-1 suggesting that the fraction contain Schiff's base and primary amine structure.

• 한국식품과학회지
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• 제11권2호
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• pp.77-80
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• 1979

녹차중(綠茶中)의 L-ascorbic acid를 정량(定量)함에 있어서 흔히 사용되고 있는 DNP법(法)을 적용할 때 불순물(不純物)에 의한 방해(妨害)여부와 그의 제거방법(除去方法)에 대하여 검토(檢討)한 결과(結果)는 다음과 같다. 즉, DNP법(法)으로 AsA를 측정시 ethyl glyoxal, diacetyl, caffeine둥 불순물질 및 아미노산(酸)들이 방해하였으며, 이들 물질을 제거하는 수단으로 브롬산화(酸化)후 o-phenylenediamine을 반응시킨 다음 chloroform으로 추출제거(抽出除去)하고 Amberlite $IR-120(H^{+})$로 아미노산(酸)을 제거(除去)한 후에 DNP법(法)으로 비색정양(比色定量)하면 L-ascorbic acid 함량(含量)을 정확히 측정(測定)할 수 있었다. 또한 분석소요시간(分析所要時間)이 TLC법(法)에 비해서 현저히 단축되고 재현성(再現性)도 좋아서 녹차중(綠茶中)의 ascorbic acid 정량(定量)에는 본법(本法)이 적합함을 확인하였다.

• 한국유기농업학회지
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• 제22권4호
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• pp.825-839
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• 2014

Antioxidant activity is important for reducing oxidative stress that causes various metabolic disorders. Metabolic disorders are highly related to loss of productivity in livestock. Therefore, development of effective antioxidant compounds originating from plants is important for organic agriculture. Phenolic compounds in edible plants are regarded as major components relevant to antioxidant activity. The present study investigated the changes in antioxidant activity and phenolic compound profiles of Aronia (Aronia meloncarpa) by fermentation using different strains of Leuconostoc mesenteroides. A total of 5 strains of L. mesenteroides were used as starter cultures and their ${\beta}$-glucosidase activities were measured. A total of 6 experiment runs were prepared, one for control (uninoculated) and the others (inoculated) for treatments. For biological activity, antioxidant and antibacterial activities were measured. For phenolic compound profiling, TLC and HPLC analysis were performed. The strains of KACC12313 and KACC12315 showed greater enzyme activity than others. Treatment with KCCM35046 showed strong and broad antibacterial activity against to Listeria monocytogenes. Treatments with KCCM35046 and KACC12315 showed the highest total polyphenol content. The highest antioxidant activity was found in KACC12315 treatment. No remarkable alteration was found in thin layer chromatography (TLC) analysis. In phenolic compound profiling analysis, KCCM35046 showed notable alteration in compound area ratio compared to others and also showed the highest caffeic acid content. In chlorogenic acid, treatments with KCCM35046 and KACC12315 showed great content than others. Treatment with KACC12315 showed the greatest content of trans-ferulic acid. As a result of relative performance indexing analysis, L. mesenteroides KCCM35046 and KACC12315 were selected as the best strain for the fermentation of Aronia.