• Journal of Life Science
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• v.15 no.3 s.70
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• pp.415-422
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• 2005

library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

• Korean Journal of Food Science and Technology
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• v.47 no.3
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• pp.380-387
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• 2015

The anti-amnesic effect of Artemisia argyi H against trimethyltin (TMT)-induced learning and memory impairment and its neuroprotective effect against $H_2O_2$-inducedoxidative stress were investigated. Cognitive behavior was examined by Y-maze and passive avoidance test for 4 weeks, which showed improved cognitive functions in mice treated with the extract. In vitro neuroprotective effects against $H_2O_2$-induced oxidative stress were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide and lactate dehydrogenase (LDH) assays. A. argyi H. extract showed protective effects against $H_2O_2$-induced neurotoxicity; moreover, LDH release into the medium was inhibited. Finally, high-performance liquid chromatography (HPLC) analysis showed that eupatilin and jaceosidin were the major phenolic compounds in A. argyi H. extract. These results suggest that A. argyi H. could be a good source of functional substances to prevent neurodegenerative diseases.

• Journal of Life Science
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• v.27 no.2
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• pp.225-232
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• 2017

This study aimed to evaluate several biological activities of Pharbitis nil and to isolate an anticancer agent from its methanol extract. Pharbitis nil seeds were extracted with methanol (PNM). Then, PNM was fractionated into solvent layers such as ethyl acetate fraction (PNE), butanol fraction (PNB), and water fraction (PNW). The biological activities of the fractions were analyzed for tyrosinase inhibition, lipase inhibition, DPPH-free radical scavenging, and cell growth inhibition. PNM showed strong growth inhibition of prostate cancer PC-3 cells. PNM was subjected to Diaion HP-20 and eluted stepwise with 50%, 80%, and 100% methanol. Then, for activity-guided fraction, each fraction was analyzed for growth inhibition of prostate cancer PC-3 cells by using an MTT assay. Because the 100% fraction showed significantly strong inhibitory activity, the fraction was further separated in the reverse phase C18, which was eluted with 80% and 90% methanol. The 90% fraction was further subjected to Sephadex LH-20 using a mobile solvent of 100% methanol. Finally, the compound PN was partially purified for HPLC analysis. PN showed cell growth inhibitory activity and induced the apoptosis and cell cycle arrest of prostate cancer PC-3 cells, as measured by flow cytometry. The results together suggest that Pharbitis nil possesses various biological activities, especially the inhibitory activity for the proliferation of prostate cancer PC-3 cells, suggesting the possibility of its use as an anticancer agent.

• Journal of Life Science
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• v.27 no.11
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• pp.1276-1289
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• 2017

This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

• Korean Journal of Agricultural Science
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• v.22 no.2
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• pp.188-196
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• 1995

To obtain useful mould strain in soybean fermentation industry, we collected conventional Meju all over the Korea and tested existance of aflatoxin in Meju collected. Also, we measured distribution of microorganism and enzyme activity of Meju and isolated Aspergillus oryzae O4-5 as a industrially useful mould. The results obtained were summarized as follows: 1. Aflatoxin was not detected by EZ-Screen Test Kit and HPLC analysis in various Meju sample collected all over the Korea from 1994.12 to 1995.2. 2. Colony forming units(CFU) of mould, yeasts, aerobe, and anaerobe were $1.3{\times}10^4-2.8{\times}10^6$, $1{\times}10^2-1.5{\times}0^6$, $2.0{\times}10^7-8.0{\times}10^8$ and $3.0{\times}10^6-7.3{\times}10^8$ per gram of Meju, respectively, indicating that CFU inter Meju samples were varied with big difference. 3. The activities of ${\alpha}$-amylase and gluco-amylase were 5-80 units and 2-34 units per g of Meju, respectively. It was shown that enzyme activity was varied depending on where the Meju was collected. 4. The activities of acidic, neutral and alkaline protease were 5-33 units, 5-302 units and 5-363 units per g of Meju, respectively. Acidic protease of Improved Meju made by D-Company was higher than that of conventional Meju as 66 units per g of Meju. 5. CNU O4-5 strain selected as a noble strain could produce amylase and protease in high level, and identified as a strain that belongs to Aspergillus oryzae. 6. The activities of acidic and neutral protease of Aspergillus oryzae CNU O4-5 strain isolated were about 10% higher than those of Aspergillus oryzae JM which has been used in soybean fermentation industry. Amylase activity of CNU O4-5 strain was similar to Aspergillus oryzae JM.

• Journal of the Korean Applied Science and Technology
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• v.37 no.3
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• pp.448-462
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• 2020

Dietary nutrition is a critical lifestyle factor that can reduce the risk of future cognitive impairments caused by dementia. Accumulating evidence suggests that dietary supplementation with Sulforaphane may help the prevention of cognitive impairments and dementia. Thus, Sulforaphane-enriched broccoli extract would hold promise to improve cognitive impairments of dementia patients. Here, we have used broccoli extracts, prepared from broccoli cultivated in Magma Seawater, to test if the broccoli extracts can be dietary supplement to improve cognitive impairments. Magma Seawater originated from Jeju Island, Korea is unique in terms of containing high concentrations of usable minerals (Zinc, Vanadium and Germanium etc.). Broccoli, grown in Magma Seawater, would contain Sulforaphane and the extra amount of usable minerals. The chemical compositions of the broccoli extracts were analyzed using LC-Q-orbitrap to detect Sulforaphane and Glucoraphanin. Analysis method based on HPLC was developed for measurement of sulforaphane levels in the broccoli extracts. We have tested if the broccoli extracts have anti-apoptotic and anti-inflammatory effects on neuron-like SH-SY5Y cells. In addition, we examined if the broccoli extracts are able to upregulate expression of synaptic plasticity-associated proteins (BDNF and phospho-CREB) and to inhibit acetylcholine esterase (AchE) activity. We have shown that the broccoli extracts inhibited the apoptotic pathway and inflammatory responses. Finally, we present evidence showing that AchE activity was inhibited by the broccoli extracts, but expression of BDNF and phospho-CREB was upregulated. Taken together, these findings suggest that the broccoli extracts from Magma Seawater-grown broccoli would be a good source of dietary nutrition to improve cognitive impairments in the future.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.9
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• pp.1256-1264
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• 2011

Infant formula is one of the main nutritional sources for infants. In this study, the fatty acid compositions of Korean (domestic, n=8) and foreign infant formulas (n=3) were analyzed. Crude fats were extracted using the Folch method, and fatty acid compositions (total and positional) were analyzed by gas chromatography. In the fatty acid compositions of infant formulas, oleic (19.88~44.64%), palmitic (7.59~21.65%) and linoleic acids (16.72~25.87%) were the major fatty acids. Domestic infant formula products contained 35.94~56.55% total saturated fatty acid content, whereas that of foreign infant formula ranged from 34.40~42.88%. The content of monounsaturated fatty acids ranged from 20.23~44.99% in domestic products and 34.11~45.07% in foreign products. In addition, 0.17~2.57% arachidonic acid/docosahexaenoic acid and 10~13% linoleic acid/linolenic acid were detected in the analyzed products (domestic and foreign products). A small amount of trans fatty acids (0.25~1.69%) were found. In sn-2 position analysis, palmitic acids (1.84~38.74%) were detected in the analyzed formulas. Further, typical triacylglycerols in human milk, including 1,3-di-monounsaturated-2-saturated triacylglycerol, were not detected in the analyzed formulas.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1580-1586
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• 2009

This study is on the effect of oil seed by-products added to Chungkukjang. For this, we designed three cases: Chungkukjang was added in with defatted sesame flour before fermented (DSFBF), added with defatted sesame flour after fermented (DSFAF) and with no adding (control). In each case, the common ingredients and the active antioxidant ingredients were examined and compared and the effects were analyzed. According to microanalysis result, carbohydrate content Chungkukjang the DSFAF 24.97%, control 23.86%, DSFBF Chungkukjang 20.21% as compared to control and Chungkukjang DSFAF relatively low carbohydrate content. The moisture contents in DSFBF (55.98%) or DSFAF (52.83%) were higher than that in control (48.89%). Chungkukjang crude ashes in DSFBF (1.48%) or DSFAF (2.41%) were much lower than in control (6.45%). The proportions of crude lipid in DSFBF (3.30%) or DSFAF (3.93%) were higher than in control (1.77%) by about 2%. As for crude protein, the percentage in DSFAF (15.86%) was lower than that of DSFBF (19.03%) or of control (19.03%). There was no meaningful difference in biological activity measurement as total phenolic contents were 1.26 mg/mL in DSFBF, 1.14 mg/mL in DSFAF and 1.26 mg/mL in control. But electron donating ability was meaningfully more active in DSFBF (21.30%) than in control (20.24%). The superoxide dismutase (SOD)-like activity in DSFBF (68.48%) was twice higher than in control (34.01%), which may imply that DSFBF contain some ingredients that can scavenge superoxide anion radically. In hydroxyl radical scavenging activity, DSFAF scores 96.87%, which is the highest with 96.40% in DSFBF and 95.73% in control. Relative antioxidative effects in DSFBF was 47.92%, which is comparable to 47.06% in control. As a result of extraction and quantitative HPLC analysis of sesamin and sesamolin extracted from the samples, DSFBF contained 3.04$\pm$0.21 mg/g of sesamin, which is meaningfully higher than 2.41$\pm$0.14 mg/g in DSFAF. Content of sesamolin was higher in DSFBF (1.36$\pm$0.09 mg/g) than DSFAF (1.12$\pm$0.07 mg/g) or in control. We can conclude that biologically active and effective ingredients could be found more in DSFBF than in DSFAF or in control. This study conveys not only the meaning that oil seed by-products can be used as an ingredient for making Chungkukjang functional food, but also the possibility that oil seed by-products themselves could become excellent functional food.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.4
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• pp.459-464
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• 2017

Biphenyl is used as an intermediate in the production of crop protection products, a solvent in pharmaceutical production, and as a component in the preservation of citrus fruits in many countries. Biphenyl is not authorized for use and also does not have standards or specifications as a food additive in Korea. National and imported food products are likely to contain biphenyl. Therefore, control and management of these products is required. In this study, a simple analytical method was developed and validated using HPLC to determine biphenyl in food. These methods are validated by assessing certain performance parameters: linearity, accuracy, precision, recovery, limit of detection (LOD), and limit of quantitation (LOQ). The calibration curve was obtained from 1.0 to $100.0{\mu}g/mL$ with satisfactory relative standard deviations (RSD) of 0.999 in the representative sample (orange). In the measurement of quality control (QC) samples, accuracy was in the range of 95.8~104.0% within normal values. The inter-day and inter-day precision values were less than 2.4% RSD in the measurement of QC samples. Recoveries of biphenyl from spiked orange samples ranged from 92.7 to 99.4% with RSD between 0.7 and 1.7% at levels of 10, 50, and $100{\mu}g/mL$. The LOD and LOQ were determined to be 0.04 and $0.13{\mu}g/mL$, respectively. These results show that the developed method is appropriate for biphenyl identification and can be used to examine the safety of citrus fruits and surface treatments containing biphenyl residues.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.4
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• pp.517-524
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• 2011

Eleven polyphenolic compounds, including procatechuic and chlorogenic acids, (+)-dihydroquercetin, rutin, isoquercitrin, quercitrin, (+)-dihydrokaempferol, trans-resveratrol, moracin, quercetin and 4-prenylmoracin were isolated and purified from the methanolic extract of defatted mulberry seed residue by a series of column chromatography including silica gel, Sephadex LH-20, and ODS-A, and their chemical structures were identified by spectral analysis. The antioxidant activities of the eleven isolated polyphenolic compounds were measured spectrophotometrically using DPPH radical. Among the eleven polyphenolic compounds tested, rutin ($IC_{50}=20.2\;{\mu}M$), isoquercitrin ($IC_{50}=22.5\;{\mu}M$), quercitrin ($IC_{50}=24.6\;{\mu}M$), quercetin ($IC_{50}=27.8\;{\mu}M$), (+)-dihydroquercetin ($IC_{50}=28.9\;{\mu}M$), and chlorogenic acid ($IC_{50}=30.6\;{\mu}M$) exhibited stronger antioxidant activity than L-ascorbic acid ($IC_{50}=31.5\;{\mu}M$) and ${\alpha}$-tocopherol ($IC_{50}=52.3\;{\mu}M$), whereas procatechuic acid ($IC_{50}=68.2\;{\mu}M$) showed lower activity. In addition, (+)-dihydrokaempferol ($IC_{50}=33.8\;{\mu}M$), trans-resveratrol ($IC_{50}=36.2\;{\mu}M$), moracin ($IC_{50}=47.6\;{\mu}M$), and 4-prenylmoracin ($IC_{50}=48.2\;{\mu}M$) exhibited moderate antioxidant activity. Furthermore, levels of the eleven polyphenolic compounds from three different types of mulberry seeds were quantified by HPLC, and their contents were as follows: rutin (311~60.0 mg/100 g)> quercitrin (7.2~34.2 mg/100 g)> (+)-dihydroquercetin (13.2~33.1 mg/100 g)> quercetin (15.8~19.5 mg/100 g)> 4-prenylmoracin (10.5~43.3 mg/100 g)> isoquercitrin (5.8~15.4 mg/100 g)> chlorogenic acid (0.0~15.3 mg/100 g)> moracin (4.7~7.2 mg/100 g)> procatechuic acid (0.0~11.6 mg/100 g)> (+)-dihydrokaempferol and trans-resveratrol (<0.1 mg/100 g). The 'Daesungppong' mulberry seeds among the three cultivars had higher flavonoid contents, such as rutin and quercetin derivatives, while the 'Iksuppong' seeds had the highest contents of phenolic acids and moracin derivatives. 'Cheongilppong' had lower amounts of polyphenolic compounds than the other two mulberry seeds. These results indicate that mulberry seeds containing antioxidant polyphenolic compounds may be potentially useful sources of anti-diabetic, anti-hypertensive, and anti-aging agents for functional foods and cosmetics.