• 생명과학회지
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• 제9권1호
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• pp.111-120
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• 1999

우리 고유의 전통 발효 음식인 김치로부터 bacteriocin 생성능이 우수한 Lactococcus sp. J-105균주를 분리하였다. Lactococcus sp. J-105의 생육 및 bacteriocin 생성에 대한 최적 조건을 검토한 결과 bacteriocin 생성은 대수 증식기 말기에서 정지기 초기에 가장 많이 생성되었다. Lactococcus sp. J-105의 생육 및 bacteriocin 생성 최적 탄소원 및 질소원은 각각 maltose와 polypeptone이었다. 균의 생육 및 bacteriocin 생성 최적 온도는 $25^{\circ}C$였으며 생육 최적 pH는 pH 8 부근이었다. bacteriocin 생성 또한 pH 8에서 최고를 나타내었다. 한편 NaCl은 오히려 균체 증식과 bac-tericoin 생성을 저해하였다. Lactococcus sp. J-105가 생산하는 bacteriocin의 pH 및 열에 대한 안정성을 검토한 결과 pH 2~4, $121^{\circ}C$ 15분에서 각각 안정하였다. 또한 J-105의 항균 spectrum을 검토한 결과 Lactobacillus sp. Leuconostoc s 그리고 Bacillus subtilis 등의 Gram 양성균에 대해서 항균활성 나타내었으며 Gram 음성균인 Acetobacter acid에 대해서도 항균 활성을 나타내었으나 E. coli에 대해서는 항균 활성을 나타내지 않았다. 본 bacteriocin은 trypsin, protease와 같은 peptide 분해효소에 대해서는 안정한 반면, pepsin에 의해서는 활성이 상실되는 특이한 성질을 나타내었다

• Journal of Applied Biological Chemistry
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• 제63권1호
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• pp.103-110
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• 2020

Objective: The objective of our study was to investigate anti-cancer effects of Danggui Liuhuang Decoction extract bioconverted by protease liquid coenzyme of Aspergillus kawachii (DLD-BE), compared to a non-bioconverted DLD extract (DLD-E) and determine the underlying mechanisms. Methods: DLD-E and DLD-BE were evaluated for their ability to modulate these signaling pathways and suppress the proliferation of human colorectal cancer (CRC) cells, HCT-116, LoVo, and HT-29. The anti-cancer effects of DLD-E and DLD-BE were measured by using proliferation and migration assays, cell cycle analysis, Western blots, and real-time PCR. Results: In this study, treatment with DLD-E and DLD-BE at concentrations of 25-100 ㎍/mL inhibited proliferation and migration in human CRC cells. DLD-BE induced apoptotic cell death and decreased COX-2 expression in HT-29 cells. The mechanisms of action included modulation of the AKT and extracellular-signal-regulated kinase signaling cascades along with inhibition of COX-2 expression. The results demonstrate novel anti-cancer mechanisms of DLD-BE against the growth of human CRC cells. Thus, we propose that DLD-BE can be developed as a more potent supplement to inhibit colorectal tumor growth and intestinal inflammation than DLD-E.

• 한국작물학회지
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• 제47권
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• pp.95-106
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• 2002

Soybean is one of the most important sources of protein and oil in the world. Recently, emphasis has been laid on the chemical composition of soybean seeds for the processing soybean foods. Improvement of soybean components has been expected to improve food-processing quality for the processed soybean products such as soymilk and various edible ingredients as well as fur the traditional soyfoods. In Korea, soybean breeding research programmes have been focused on the quality of the products derived from soybean with yield stability, and some new modified soybean varieties haying good food-processing quality were developed recently. So the efforts of establishing standard and standardization of products in soybean are important. Three main categories should be considered in view of soybean seed quality; the marketing value such as grain size, shape, and appearance; the eating and processing value such as dehulled ratio, water absorption rate, and benny flavor; the nutritional value such as protein, lipid, and carbohydrate contents. And the new frontiers in research are looking at the functional nutrients in soybeans and how to improve them. In case marketing value, mainly the appearance is evaluated, therefore, each country has an application of standard related to quality. Each determination of standard class, heat-damaged kernels, splits, and soybeans of other colors is made on the basis of the grain when free from foreign materials. But processing value and nutritional value for standardization were not studied in detail till now. In addition, soybean has potential roles in the prevention and treatment of chronic diseases, most notably cancer, osteoporosis, and heart disease. The functional nutrients include a protease inhibitor, phytic acid, saponins, and isoflavones, etc.. It is believed that standardization of soybean quality should perform to overcome the difficulties, relatively high price of domestic soybean products has weakened the competitive power, in the market related to WTO. So, we should focus on further research into the evaluation and establishment of quality-standard in soybean.

• KSBB Journal
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• 제32권1호
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• pp.71-82
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• 2017

To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.

• 대한한의학회지
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• 제23권1호
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• pp.24-36
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• 2002

Objectives: The chemotherapeutic potential of Gagamgilgyung-tang for the treatment of human lung cancer, the antitumorigenic effects of Gagamgilgyung-tang on the proliferation and apoptosis of human lung cancer cell line A427 were investigated using molecular biological approaches, Methods: To determine Gagamgilgyung-tang concentrations which do not evoke cytotoxic damage to the cell line, cell viability was examined by MTT assay. To prove Gagamgilgyung-tang's antitumorigenic potential to human lung cancer, [3H]thymidine incorporation assay, trypan blue exclusion and Cpp32 protease activity assays and quantitative RT-PCR analysis were examined. Results: While A427 cells treated with $0.1-2.0{\mu\textrm{g}}/ml$ of Gagamgilgyung-tang showed no recognizable effect, marked reductions of cell viability were detected at concentrations over $5.0{\;}\mu\textrm{g}/ml$. DNA replication of A427 cells was inhibited by Gagamgilgyung-tang in a dose-dependent manner and Gagamgilgyung-tang induced the G1 cell cycle arrest through inhibition of DNA replication. Gagamgilgyung-tang triggered apoptotic cell death of A427 and enhanced the apoptotic sensitivity of the cells that were injured by a DNA damage-inducing chemotherapeutic drug etoposide. Gagamgilgyung-tang induces expression of growth-inhibiting genes such as p53 and p21/Wafl whereas it inhibited expression of growth-promoting genes such as c-Myc and Cyclin D1. Expression of a representative apoptosis-inducing gene Bax was also found to be induced by Gagamgilgyung-tang while apoptosis-suppressing Bcl-2 expression was not changed. Conclusions: Gagamgilgyung-tang could suppress the abnormal growth of tumor cells by suppressing the survival of genetically altered cells via induction of apoptosis. This study suggests that Gagamgilgyung-tang might have an antitumorigenic potential to human lung cancer cells, which might be associated with its growth-inhibiting and apoptosis-inducing properties.

• 한국응용과학기술학회지
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• 제33권1호
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• pp.176-185
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• 2016

연산오계는 오래전부터 건강기능 증진 및 치료 효능이 높은 것으로 알려져 왔다. 최근 건강기능식품 소재로 기능성 펩타이드 효능이 알려짐에 따라, 연산오계 다리육으로부 올리고 펩타이드 최적 생산 공정 및 생성물 특성에 대하여 연구를 수행하였다. 최적 효소가수 분해 공정 표면반응 분석을 이용하여 수행하였다. 최적 공정 조건을 확립하기 위해서 온도 (40, 50, $60^{\circ}C$), pH (pH 6.0, 7.0, 8.0), 효소 (1, 2, 3%) 범위에서 수행을 하였다. 생성물에 대한 가수분해도, 유리아미노산, 분자량 분포를 분석하였다. 효소 가수분해 최적 온도는 $58^{\circ}C$, pH 7.5, 효소의 농도는 3% 이었다. 최적 조건에서 2 시간 효소 가수분해를 한 결과 75-80% 이었다. 유리 아미노산 총량은 168.131 mg/100 g 이었다. 분자량를 MALDI-TOF 으로 분석을 한 결과 90% 이상이 300-1,000 Da 분포를 보여주었다.

• Genomics & Informatics
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• 제14권3호
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• pp.104-111
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• 2016

Zika virus (ZIKV) is a mosquito borne pathogen, belongs to Flaviviridae family having a positive-sense single-stranded RNA genome, currently known for causing large epidemics in Brazil. Its infection can cause microcephaly, a serious birth defect during pregnancy. The recent outbreak of ZIKV in February 2016 in Brazil realized it as a major health risk, demands an enhanced surveillance and a need to develop novel drugs against ZIKV. Amodiaquine, prochlorperazine, quinacrine, and berberine are few promising drugs approved by Food and Drug Administration against dengue virus which also belong to Flaviviridae family. In this study, we performed molecular docking analysis of these drugs against nonstructural 3 (NS3) protein of ZIKV. The protease activity of NS3 is necessary for viral replication and its prohibition could be considered as a strategy for treatment of ZIKV infection. Amongst these four drugs, berberine has shown highest binding affinity of -5.8 kcal/mol and it is binding around the active site region of the receptor. Based on the properties of berberine, more similar compounds were retrieved from ZINC database and a structure-based virtual screening was carried out by AutoDock Vina in PyRx 0.8. Best 10 novel drug-like compounds were identified and amongst them ZINC53047591 (2-(benzylsulfanyl)-3-cyclohexyl-3H-spiro[benzo[h]quinazoline-5,1'-cyclopentan]-4(6H)-one) was found to interact with NS3 protein with binding energy of -7.1 kcal/mol and formed H-bonds with Ser135 and Asn152 amino acid residues. Observations made in this study may extend an assuring platform for developing anti-viral competitive inhibitors against ZIKV infection.

• 한국동물학회지
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• 제36권3호
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• pp.347-352
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• 1993

N-ethylmaleimide는 낮은 농도에서 Myosin heavy chain의 활성화 부위에 존재하는 2개의 황화기에 선택적으로 결합하는 것으로 알려져 있다. 계 근조직에서 분리된 $Ca^2$+-의존성 단백질 분해효소, Calpain은 이와같이 알킬화된 Myosin heavy chain을 알킬화 되지 않은 것에 비하여 우선적으로 분해하는 것으로 나타났다. 또한, 황화기를 특이하게 산하시키는 KMnO$_4$가 처리된 Myosin heavy chain도 산화되지 않은 것에 비하여 훨씬 빠른 속도로 분해됨을 관찰하였다. 뿐만아니라, N-ethylmaleimide나 KMnO$_4$의 처리는 농도-의존적으로 myosin에 의한 ATP 분해를 불활성화 시키었다. 이러한 결과는 활성화 부위에 존재하는 황화기의 화학적 변형은 Myosin hsavy chain이 Calapin과 같은 세포내 단백질 분해효소에 의하여 인식되는 기구임을 시사한다.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.55-70
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• 2005

Objective: Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammnatory in the traditional Korean medicine. UD has been reported as a good enhancer for bone healing. Methods : In this experiment, we investigate the Inhibitory effects of UD on bone resorption using the bone cells culture. Different concentrations of crude extract of UD were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MIT assay. It was demonstrated that UD has potential effects on bone cells culture without any cytotoxicity. The most effective concentration of UD on bone cells were $100\;{\mu}g/ml$. Cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. Results : When mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-Kinase inhibitor, wortmannin (WT), WT prevented the osteoclast-mediated intracellular processing of Cat K. Similarly, treatment of osteoclasts-containing long bone cells with UD extracts prevented the intracellular maturation of Cat K, suggesting that UD may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose-6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and UD in the absence or presence of M6P. Inhibition of Cat K processing by WT or UD was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and UD. Conclusion : UD dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing.

• 한국식품과학회지
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• 제35권4호
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• pp.680-683
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• 2003

한약재로부터 U937 세포주를 사용한 caspase-3 유도저해물질을 탐색한 결과 소목(Caesalpiniae sappan)을 선발하였다. 소목의 메탄올 추출물로부터 silica gel column chromatography, HPLC 등을 사용하여 저해물질을 분리하였으며, ESI-MS, $^1H-NMR,\;^{13}C-NMR$ 등의 기기분석을 실시하여 brazilin이 동정되었다. 이 물질은 $IC_{50}\;4.5\;{\mu}g/mL$의 농도로 U937세포주의 etoposide에 의한 caspase-3 유도를 저해하였으며, DPPH 라디칼 소거능도 나타내었다.