• 생명과학회지
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• 제14권2호
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• pp.315-319
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• 2004

사람의 serine palmitoyltransferase(SPT, EC 2.3.1.50)에 대한 항체를 제작하기 위하여 E. coli발현 vector인 pRset vector에 SPTLC1 및 SPTLC2 유전자를 subcloning하고 BL21 (DE3)pLys cell에 발현시켰다. 포유동물의 SPT는 원핵세포의 SPT homodimer와는 달리 SPTLC1 및 SPTLC2 2개의 sub-unit로 된 heterodimer이다. Human embryo kidney cell인 HEK293 cell의 total RNA로부터 RT-PCR을 행하여 cDNA library를 얻은 다음 SPTLC1 및 SPTLC2의 특이적인 primer 들을 이용하여 PCR을 행하였다. SPTLC1 및 SPTLC2 DNA를 hexahistidine fusion 단백질을 발현시킬 수 있는 pRset vector에 cloning하여 pRsetB/SPTLC1 및 pRsetA/SPTLC2를 얻고 염기서열을 확인하였다. 재조합 plasmid를 발현세포인 BL21 cell에 형질전환시킨 다음 ampicillin 및 chroramphenicol 배지에서 선별하여 재조합세포를 얻었다. 1 mM IPTG로서 발현을 유도하였으며 세포 단백질을 SDS-PAGE로 분리한 다음 His-tag antibody로 western blotting을 행하여 SPTLC 및 SPTLC2가 발현되었음을 확인하였다.

• Preventive Nutrition and Food Science
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• 제13권3호
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• pp.196-203
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• 2008

Hwangtae, dried Alaska pollack, is a major storage product in the fish processing industry. Hwangtae is prepared by removing the internal organs and drying outdoors during the cold witner months by allowing it to thaw during the daytime and re-freeze at night under sub-zero ($-10^{\circ}C$) conditions and gradually dry from December until the next April for around 5 months from Myungtae. In this study, ground Hwangtae was hydrolyzed using two proteolytic enzymes (pepsin and alcalase) which produced five soluble active peptides from Hwangtae (yellowish dried Pollack, Theragra chalcogramma) protein. Two different peptides with strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods of Sephadex G-25 gel, ion-exchange chromatography on a Sepharose-Sephadex C-25 gel, and high-performance liquid chromatography. The isolated peptides, APO1 and APO2, were composed of 16 and 13 amino acid residues, respectively. Both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The peptide with a molecular weight less than 1,000 Daltons (APACE) obtained from enzymatic hydrolysates of Hwangtae exhibited the highest ACE inhibitory activity. The APACE peptides was composed of 4 amino acid residues (Gly-Leu-Leu-Pro). These results suggest that Hwangtae hydrolysates could be a good source of peptides with ACE inhibitory activity. Biochemical analysis indicated that two 70 kDa peptides (APG1 and APG2) isolated from the hydrolysate had gelatinoytic activity, which was shown to be a calcium dependent protease type as showed by gelatin SDS PAGE.

• 한국자원식물학회지
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• 제30권1호
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• pp.88-95
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• 2017

토양수분의 변화가 블랙쵸크베리 'Nero'의 과실 특성에 미치는 영향을 조사하여 고품질 블랙쵸크베리 재배 연구의 기초자료를 제공하고자 수행하였다. 토양수분 감소로 인한 건조스트레스는 수확기 동안 블랙쵸크베리의 과실의 과중, 당도, 안토시아닌 함량 등의 품질을 저하시켰다. 또한 건조기간이 길수록 이후 토양수분 공급시, 과실 표피 세포 크기는 감소하는 반면 아표피, 과육 세포의 크기가 증가하여 열과를 유발할 수 있다. 따라서 관수시설을 통한 일정 주기의 관수가 블랙쵸크베리 과실 생산량 및 품질 향상을 위해 필요할 것으로 생각된다.

• Archives of Pharmacal Research
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• 제28권2호
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• pp.195-202
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• 2005

Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental factors are critical players in cellular damage and aging. In order to develop a new antiphotoaging agent, this work focused on the antioxidant effects of the extract of tinged autumnal leaves of Acer palmatum. One compound was isolated from an ethyl acetate soluble fraction of the A. palmatum extract using silica gel column chromatography. The chemical structure was identified as apigenin-8-C-beta-D-glucopyranoside, more commonly known as vitexin, by spectral analysis including LC-MS, FT-IR, UV, $^{1}H-$, and $^{13}C-NMR$. The biological activities of vitexin were investigated for the potential application of its anti-aging effects in the cosmetic field. Vitexin inhibited superoxide radicals by about $70\%$ at a concentration of $100\;{\mu}g/mL$ and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals by about $60\%$ at a concentration of $100\;{\mu}g/mL$. Intracellular ROS scavenging activity was indicated by increases in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20\;mJ/cm^2$ in cultured human dermal fibroblasts (HDFs) after the treatment of vitexin. The results show that oxidation of 5-(6-)chloromethyl-2',7'-dichlo-rodihydrofluorescein diacetate ($CM-H_{2}DCFDA$) is inhibited by vitexin effectively and that vitexin has a potent free radical scavenging activity in UVB-irradiated HDFs. In ROS imaging using a confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our findings suggest that vitexin can be effectively used for the prevention of UV-induced adverse skin reactions such as free radical production and skin cell damage.

• Archives of Pharmacal Research
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• 제26권5호
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• pp.383-388
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• 2003

Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity ($IC_{50} = 13.1 \mu$ g/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and BF-3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-$\alpha and IL-1\beta$ are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-$\alpha and IL-1\beta$ production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-$\alpha and IL-1\beta$production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production.

• 동아시아식생활학회지
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• 제26권6호
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• pp.543-549
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• 2016

The objective of this study was to investigate quality properties of 'Cheonghyang' wine using different commercial yeast strains. Soluble solid content, pH level and total acidity of 'Cheonghyang' grape were $20^{\circ}Bx$, pH 3.5 and 0.66%, respectively. Total acidity ranged from 0.91~1.06% in the middle stage of fermentation and decreased to 0.77~0.82% when alcoholic fermentation finished. Alcohol content in wines ranged from 12.5% to 12.9% showing no significant difference in yeast strains. Wine fermented with Red Fruit had high volatile acid content (189.0 mg/L) whereas wine fermented with Fermivin indicated low volatile acid content (77.7 mg/L). Wines made with Montrachet, Fermivin and Aroma White had low brightness (L-value) compared to others. Results from sensory evaluation demonstrated that commercial wine yeasts, Montrachet and Fermivin, can be applied to improve sensory properties of 'Cheonghyang' wine such as aroma, acidity and transperency. On the other hand, preferences of wine fermented with EC-1118 strain containing lots of tannins and total polyphenols were significantly reduced.

• Molecules and Cells
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• 제40권5호
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• pp.355-362
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• 2017

The ${\beta}2$ integrins are cell surface transmembrane proteins regulating leukocyte functions, such as adhesion and migration. Two members of ${\beta}2$ integrin, ${\alpha}M{\beta}2$ and ${\alpha}X{\beta}2$, share the leukocyte distribution profile and integrin ${\alpha}X{\beta}2$ is involved in antigen presentation in dendritic cells and transendothelial migration of monocytes and macrophages to atherosclerotic lesions. ${\underline{R}}eceptor$ for ${\underline{a}}dvanced$ ${\underline{g}}lycation$ ${\underline{e}}nd$ ${\underline{p}}roducts$ (RAGE), a member of cell adhesion molecules, plays an important role in chronic inflammation and atherosclerosis. Although RAGE and ${\alpha}X{\beta}2$ play an important role in inflammatory response and the pathogenesis of atherosclerosis, the nature of their interaction and structure involved in the binding remain poorly defined. In this study, using I-domain as a ligand binding motif of ${\alpha}X{\beta}2$, we characterize the binding nature and the interacting moieties of ${\alpha}X$ I-domain and RAGE. Their binding requires divalent cations ($Mg^{2+}$ and $Mn^{2+}$) and shows an affinity on the sub-micro molar level: the dissociation constant of ${\alpha}X$ I-domains binding to RAGE being $0.49{\mu}M$. Furthermore, the ${\alpha}X$ I-domains recognize the V-domain, but not the C1 and C2-domains of RAGE. The acidic amino acid substitutions on the ligand binding site of ${\alpha}X$ I-domain significantly reduce the I-domain binding activity to soluble RAGE and the alanine substitutions of basic amino acids on the flat surface of the V-domain prevent the V-domain binding to ${\alpha}X$ I-domain. In conclusion, the main mechanism of ${\alpha}X$ I-domain binding to RAGE is a charge interaction, in which the acidic moieties of ${\alpha}X$ I-domains, including E244, and D249, recognize the basic residues on the RAGE V-domain encompassing K39, K43, K44, R104, and K107.

• 대한한방부인과학회지
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• 제20권3호
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• pp.81-90
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• 2007

Purpose: This study was conducted to investigate the muscle anti-fatigue effects of Allii tuberosi semen after long term administration. Materials and Methods: 4-6 weeks old ICR mice were used in this study and we administered the water soluble extracts of Allii tuberosi semen in the concentration of 1, 10 and 100mg/0.3ml to each 5 mice (ATS group) and same volume of normal saline was administered to 5 mice (control group) once a day for 90 or 120 days. After the administration we performed the swimming exercise test and the grip strength test. And we measured the concentration of glucose and the activity of lactate dehydrogenase in plasma regarded as biochemical indicator related with the muscular fatigue. Results: In swimming exercise test, all ATS groups showed increased time compared with relative control group after 90 days administration. But after 120 days adminstration 10 and 100mg/0.3ml ATS groups showed increased results. In grip strength test, all ATS group increased grip strength after 90 days administration and 10mg/0.3ml ATS group showed increased result after 120 days administration. All the results of plasma glucose didn't showed statistically significant difference after 90 and 120 days administration. In the plasma concentration of activity level of lactate dehydrogenase, 1mg/0.3ml and 10mg/0.3ml ATS groups showed decreased concentration compared with relative control group after 90 days administration. But after 120 days adminstration 1mg/0.3ml ATS groups showed decreased results. Conclusion: From this study we could find the anti-fatigue effect of Allii tuberosi semen in the muscle strength related test.

• 한국물환경학회지
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• 제37권2호
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• pp.92-102
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• 2021

In this study, the distribution characteristics according to the type of BOD and the effect of nitrogenous oxygen demand (NOD) and algal oxygen demand on BOD in three streams (Bokhacheon, Byeongseongcheon, and Gulpocheon) were evaluated. Although the BOD and NOD concentrations demonstrated a difference in the three streams, the carbonaceous BOD(CBOD)/BOD ratio was 0.75 (p=0.053, one-way ANOVA), and there was no significant difference in the three streams (r2≥0.92, p<0.0001). The NOD concentration of the Bokhacheon with high NH3-N was 1.7±1.3 mg/L, which was the highest among the three streams and showed a significant correlation with BOD. Seasonal variations in NOD in the three streams did not show a significant correlation with changes in NH3-N concentration (r2<0.28, p≥0.1789), and there was no significant difference in NOD even though NH3-N concentration in Gulpocheon was about twice that of Byeongseongcheon (p=0.870, one way ANOVA). The particulate CBOD(PCBOD)/CBOD ratio of the three streams was 0.55~0.64, and about 60% of the biodegradable organic matter was present in the particulate form. When the Chl.a concentration in the stream was more than 7 ㎍/L, the PCBOD tended to increase with the Chl.a concentration (r2=0.61, p=0.003). In the three streams, particulate NOD accounted for 81% of NOD; however, despite the large variation in NH3-N concentration (0.075~3.182 mg/L), there was no significant difference in soluble NOD(SNOD) concentration that ranged from 0.1 to 0.3 mg/L. In this study, the low contribution rate of SNOD to NOD is considered as a result of the removal of nitrifying bacteria along with the particles during the filtration process.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.97-102
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• 2021

본 연구에서는 들깨(Perilla frutescens)박으로부터 효소분해물을 제조하기 위한 효소를 선별하여 최적의 반응조건을 확립하였다. Alcalase와 Ceremix의 동시 처리가 들깨박의 단백질과 탄수화물의 가용화에 효과적이었으며, 효소 사용량은 들깨박 중량의 2% (w/w), pH는 7.0, 반응 시간은 2시간이 적당하였다. 최적의 반응조건에서 들깨박을 Alcalase와 Ceremix로 처리한 결과 환원당, 가용성 단백질 및 총 폴리페놀 함량이 크게 증가하였다. 유리 라디칼과 양이온 라디칼에 대한 소거활성으로 확인한 결과 들깨박 효소분해물의 항산화 활성은 대조군에 비해 우수하였다. 또한 젖산균인 Leuconostoc mesenteroides 310-12 균주를 들깨박 효소분해물에서 배양한 결과 대조군에 비해 생육과 산의 생성이 우수하였다. 결론적으로 들깨박 효소분해물은 항산화 생리활성 소재로서뿐만 아니라 유산균 배양 배지로서도 활용 가능성이 있음을 확인하였다.