• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.69-75
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• 2013

Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.

• Korean Journal of Pharmacognosy
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• v.45 no.3
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• pp.209-213
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• 2014

The anti-melanogenic substance was isolated from the root of Angelica gigas Nakai by silica gel column chromatography, preparative HPLC and TLC. As a result of the structure analysis by mass, $^1H$-NMR, and $^{13}C$-NMR spectrometry, the compound was identified as demethylsuberosin. Demethylsuberosin reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner and decreased to about 74% at a concentration $5{\mu}g/ml$. Demethylsuberosin inhibited the expression in microphthalmia associated transcription factor (MITF), tyrosinase, tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2) in melanocytes. These results suggest that the whitening activity of demethylsuberosin may be due to the inhibition of the melanin synthesis by down-regulation of MITF, tyrosinase, TRP-1 and TRP-2 expression. Thus, our results provide evidence that demethylsuberosin might be useful as a potential skin-whitening agent.

• Natural Product Sciences
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• v.16 no.2
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• pp.80-87
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• 2010

High performance liquid chromatography (HPLC) analysis on the MeOH extract of Ligularia stenocephala leaves identified six caffeoylquinic acids, viz. 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoyl-mucoquinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, and 3-Ocaffeoylquinic acid using standard compounds, and determined the quantity of each extract. Percentage of total caffeoylquinic acids of the MeOH extract and its BuOH fraction were calculated as $67.83{\pm}3.79%$ and $94.52{\pm}1.84%$, respectively. Since the caffeoylquinic acid-rich MeOH extract exhibited a potent peroxynitrite-scavenging effect in vitro ($IC_{50}=0.87{\pm}0.33\;{\mu}g/ml$ (mean $\pm$ SEM)), the experiment was designed to identify whether or not that extract has an anti-obesity effect on rat obesity induced by high fat diet. Oral administration of the MeOH extract and its BuOH fraction abundant in caffeoylquinic acid decreased the rat body weight to the level of untreated group and decreased abdominal fat pad weight. The atherogenic index and thiobarbituric acid-reactive substance (TBARS) values were restored by treatment, indicating that the caffeoylquinic acid-rich extract probably inhibited hyperlipidemia and oxidative stress caused by high fat diet. These results suggest that L. stenocephala in vegetable form or its caffeoylquinic acid-rich fraction (BuOH fraction) as an agent can be used for treatment or prevention of obesity.

• Korean Journal of Pharmacognosy
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• v.48 no.2
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• pp.160-165
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• 2017

Capsicum annuum L. (CA) is reported to be the most widely cultivated species of the Capsicum throughout the world. Its fruit and the seeds have been used as spice after drying and grinding. It is reported that the fruit and seeds contain capsaicinoid such as capsaicin and dihydrocapsaicin, carotenoid such as capsanthin and ${\beta}$-carotene with its anti-tumor effect, anti-obesity effect, anti-diabetic action. Present study, we isolated two lignans, icariside $E_5$ and vanilloyl icariside $E_5$ from the seed of CA and validation and content determination were confirmed using High-Performance Liquid Chromatography (HPLC). As a result, the contents of the lignans, icariside $E_5$ and vanilloyl icariside $E_5$ were 0.28%, 0.085%, respectively in the extract.

• Korean Journal of Medicinal Crop Science
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• v.28 no.4
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• pp.260-275
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• 2020

Background: We investigated the antioxidant, anti-wrinkles, whitening, and moisturizing properties and amounts of phenolic compounds of ethanol extracts from flowers of 10 resource plants from Namwon and Mt. Jiri., Korea. Methods and Results: We measured antioxidant efficacy based on the total polyphenol, and total flavonoid content, and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. We evaluated the inhibitory effect on melanin synthesis and tyrosinase activity for the whitening effect. Furthermore, we analyzed the elastase and matrix metalloproteinase-1 (MMP-1) inhibition activity for anti-wrinkle capacity. To evaluate the moisturizing effect, we examined hyaluronan synthase (HAS) mRNA expression. In addition, the 19 phenolic compounds were detected using high performace liquid chromatography (HPLC). Among the 10 flowers, the antioxidant effect was high in the order of Rosa multiflora, Nelumbo nucifera, and Elsholtzia splendens. Whitening effect was high in the order of N. nucifera, R. multiflora, and Dendranthema zawadskii. As for the anti-wrinkle property, N. nucifera was the most effective followed by R. multiflora. Taraxacum coreanum was the best for moisturizing effect, followed by D. zawadskii, and E. splendens. Seven phenolic compounds were detected in the extracts of the 10 flowers. Conclusions: Overall, the extracts of five flowers extracts showed strong potential as antioxidant, whitening, anti-wrinkle, and moisturizing functional cosmetic agents.

• The Journal of Internal Korean Medicine
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• v.22 no.3
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• pp.341-351
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• 2001

Object: Acori graminei rhizoma(AGR) extract is clinically used to treat the cerebral ischemia in Korea. The present study was undertaken to study the neuroprotective effect of AGR extract in middle cerebral artery occlusion(MCAO) rats. Methods: Changes of extracellular levels of glutamate, aspartate, GABA, glycine, taurine, tyrosine, alanine in striatum were collected at 20 minutes interval by in vivo microdialysis and then analyzed by HPLC(high performance liquid chromatography) in rats subjected to permanent focal cerebral ischemia induced by 2 hours of MCAO. AGR extract was orally administrated before MCAO. Different animals were used for measurement of cerebral infarction volume induced by 24 hours of MCAO with TTC staining and image analysis. Result : The infarction volume was decreased and focal cerebral ischemia - induced increase of extracellular glutamate, aspartate, and tyrosine were inhibited after the treatment of AGR extract. On the other hand, the increase of glycine and alanine not but GABA and taurine were enhenced after the treatment of AGR extract. Conclusion: These results suggest that AGR extract can playa role in protecting against cerebral ischemia by regulating extracellular levels of excitatory and inhibitory amino acid neurotransmitters.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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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.

• Korean Journal of Food Science and Technology
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• v.12 no.2
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• pp.126-132
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• 1980

An attempt was made to remove red color and capsaicin which were contaminated in crude oil extracted from red pepper seeds with n-hexane. Both clay bleachidg and deodorization were responsible for the removal of red color. Clay bleaching was also the most effective for the removal of capsaicin, which was removed completely with 4 % clay. Capsaicin in oil was adsorbed onto the clay particles exponentially and it followed Freundlich equation : ${\frac{x}{m}}=1.1\;c^{0.31}$, where x is amount of capsaicin adsorbed (mg/ml), m is amount of clay used in bleaching (%) and c is amount of residual capsaicin (mg/ml). Neutral lipid of the seed oil consisted of 95.8 % triglyceride, 1.55 % sterol ester, 1.45 % monoglyceride, 0.78 % free fatty acids, 0.2 % diglycerides and 0.15 % sterol. Major components of fatty acids in the red pepper seed oil were linoleic acid (78.79 %), palmitic acid (15.27 %) and linolenic acid content was only 0.63 % by high performance liquid chromatography.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.4
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• pp.582-587
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• 1997

The changes of ascorbic acid(AsA) and dehydroascorbic acid(DHAA) contents in distilled water, carrot juice, and carrot+vegetable juices under different incubation time and temperature were determined by using high performance liquid chromatography. AsA in carrot juice was more stable than that in distilled water. AsA contents in distilled water and carrot juice were gradually reduced in a time and temperature dependent manner. AsA contents in carrot juice and carrot+ vegetable juices stored in refrigerator(4$^{\circ}C$) for 2 and 24 hours appeared to decrease, but the DHAA contents in all samples increased. Total vitamin C(AsA+DHAA) contents in carrot juice and carrot +vegetable juices remained with the high residue values of 90~97% after incubating at 4$^{\circ}C$ for 2 and 24 hours.