• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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• pp.51-56
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• 1999

Plastids, which are organelles unique to plant cells, bear their own genome that is organized into DNA-protein complexes (nucleoids). Regulation of gene expression in the plastid has been extensively investigated because this organelle plays an important role in photosynthesis. Few attempts, however, have been made to characterize the regulation of plastid gene expression at the chromosomal structure, using plastid nucleoids. In this report, we summarize the recent progress in the characterization of DNA-binding proteins in plastids, with special emphasis on CND41, a DNA binding protein, which we recently identified in the choloroplast nucleoids from photomixotrophically cultured tobacco cells. CND41 is a protein of 502 amino acids which consisted of a transit peptide of 120 amino acids and a mature protein of 382 amino acids. The N-terminal of the 'mature' protein has lysine-rich region which is essential for DNA-binding. CNA41 also showed significant identities to some aspartyl proteases. Protease activity of purified CND41 has been recently confirmed and characterized. On the other hand, characterization of accumulation of CND41 both in wild type and transgenic tobacco with reduced amount of CND41 suggests that CND41 is a negative regulator in chloroplast gene expression. Further investigation indicated that gene expression of CND41 is cell-specifically and developmentally regulated as well as sugar-induced expression. The reduction of CND41 expression in transgenic tobacco also brought the stunted plant growth due to the reduced cell length in stem. GA3 treatment on apical meristem reversed the dwarf phenotype in the transformants. Effects of CND41 expression on GA biosynthesis will be discussed.

• 한국환경과학회지
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• 제25권5호
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• pp.645-654
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• 2016

For the purpose of reuse the wasted by-products from the skate process to the health functional food or medicinal material, chondroitin sulfate was extracted from the skate cartilage with the method of hydrolysis with protease enzyme, and the contents of chondroitin sulfate and hydrolyzed protein were measured qualitatively and quantitatively. The effects of chondroitin sulfate on body weight or liver weight changes, hepatotoxicity elimination and anti-inflammatory actions were measured from in vivo test with feed-treated mice. From the hydrolytic extraction of skate cartilage with the mixture of 1% alcalase and 1% protease for 4 hours, the extraction yield of chondroitin sulfate was about 32.55%. The content and molecular weight of chondroitin sulfate was 26.63% and $2.85{\times}10^5Da$., respectively and the content ratio of chondroitin sulfate to protein was measured to 1 to 2.76 with gel permeation chromatography. For the odor component, trimethylamine decreased about 30% but almost not ammonia from chondroitin sulfate with the treatment of activated carbon. From the feeding chondroitin sulfate to mice, the control effect of body and liver weights decrease was measured, anti-inflammatory action and hepatotoxicity elimination action were also measured. From these results, process operation conditions for manufacturing of chondroitin sulfate were suggested.

• Animal cells and systems
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• 제15권2호
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• pp.139-146
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• 2011

Secretory leukocyte protease inhibitor (SLPI) plays an important role in promoting the invasion and metastasis of a range of cancer cells. However, there are no reports of the expression and function of SLPI in oral carcinoma cells. In this study, the oral carcinoma cell line KB was used to determine whether SLPI affects the proliferation, migration and invasion of oral carcinoma cells. RT-PCR and Western blotting revealed high levels of endogenous SLPI expression in KB cells as well as a strong increase in SLPI secretion after wounding compared to immortalized normal oral keratinocytes (INOK). The wound healing assay revealed more migration of KB cells than INOK cells, and the SLPI treatment increased the migration of KB cells. KB cell proliferation was increased significantly by the SLPI protein but decreased by SLPI-siRNA. SLPI strongly increased the migration and invasion of KB cells. On the other hand, SLPI-siRNA decreased the migration and invasion of KB cells. This suggests that SLPI plays an important role in the metastasis of oral carcinoma cells.

• Molecules and Cells
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• 제44권9호
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• pp.688-695
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• 2021

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become a global health concern. Various SARS-CoV-2 vaccines have been developed and are being used for vaccination worldwide. However, no therapeutic agents against coronavirus disease 2019 (COVID-19) have been developed so far; therefore, new therapeutic agents are urgently needed. In the present study, we evaluated several hepatitis C virus direct-acting antivirals as potential candidates for drug repurposing against COVID-19. Theses include asunaprevir (a protease inhibitor), daclatasvir (an NS5A inhibitor), and sofosbuvir (an RNA polymerase inhibitor). We found that asunaprevir, but not sofosbuvir and daclatasvir, markedly inhibited SARS-CoV-2-induced cytopathic effects in Vero E6 cells. Both RNA and protein levels of SARS-CoV-2 were significantly decreased by treatment with asunaprevir. Moreover, asunaprevir profoundly decreased virion release from SARS-CoV-2-infected cells. A pseudoparticle entry assay revealed that asunaprevir blocked SARS-CoV-2 infection at the binding step of the viral life cycle. Furthermore, asunaprevir inhibited SARS-CoV-2 propagation in human lung Calu-3 cells. Collectively, we found that asunaprevir displays broad-spectrum antiviral activity and therefore might be worth developing as a new drug repurposing candidate for COVID-19.

• 한국식품위생안전성학회지
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• 제33권6호
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• pp.516-520
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• 2018

박테리오신은 미생물이 생산하는 단백질성의 항균물질이다. 본 연구에서는 Enterococcus faecium CJNU 2008 균주로부터 생산되는 박테리오신에 대한 일부 특성을 규명하였다. 부분 정제 박테리오신은 열처리($100^{\circ}C$ 30분, $121^{\circ}C$ 15분) 및 유기용매(메탄올, 에탄올, 아세톤, 아세토니트릴, 클로로포름)에 대한 안정성이 우수하였으며 효소처리의 경우 Lipase와 ${\alpha}-amylase$에 대해서는 안정하였으나 Protease 처리에서 활성이 소실되었다. 이는 E. faecium CJNU 2008균주가 생산하는 항균물질이 단백질성의 박테리오신임을 추가적으로 증명하는 것이다. 병원성 세균인 Listeria monocytogenes 균주를 지시균으로 사용했을 때 박테리오신은 살균(bactericidal)의 작용양상을 보였다. Tricine-SDS-PAGE를 이용한 박테리오신의 분자량은 6.5 kDa 이하로 확인되었다. 부분 정제된 박테리오신을 이용하여 HPLC법을 활용한 정제를 수행하였으며 크로마토그램 상에서 단일 피크를 얻었을 수 있었다. 앞으로 정제된 박테리오신은 생화학적 분석 등에 활용할 계획이다.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.462-470
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• 2022

The addition of dietary proteases (PRO) to weaner diets hydrolyzes soybean-based anti-nutritive factors and improves weaned pig's dietary digestibility and growth performance. Therefore, this study explores the effects of PRO in a lower crude protein (CP) level diet than that in a commercial diet on the growth performance, nutrient digestibility, and intestinal morphology of weaned pigs. A total of 90 weaned pigs were randomly assigned to 3 dietary treatments with 6 pigs per pen and 5 replicated pens per treatment using a randomized complete block design (block = body weight [BW]): 1) a commercial weaner diet as a positive control (PC; phase1 CP = 23.71%; phase2 CP: 22.36%), 2) lower CP diet than PC as a negative control (NC; 0.61% less CP than PC), and 3) an NC diet with 0.02% PRO. Pigs fed PC and PRO had higher (p < 0.05) final BW, average daily gain, and/or gain to feed ratio for the first three weeks and the overall experimental period than NC. The PC and PRO groups had greater (p < 0.05) apparent ileal digestibility of dry matter, CP, and energy than the NC group. Moreover, pigs fed PC and PRO increased (p < 0.05) apparent total tract digestibility of CP compared with those fed NC. In addition, the PRO group had a higher number of goblet cells than the PC and NC groups. However, pig fed PC and PRO increased (p < 0.05) villus height and height to crypt depth ratio in the ileum compared with those fed NC. In conclusion, PRO supplementation in a commercial weaner diet with low CP levels improves growth rate and nutrient digestibility by modulating the intestinal morphology of weaned pigs.

• Journal of Nutrition and Health
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• 제42권4호
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• pp.397-405
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• 2009

본 연구에서는 식품 중의 엽산을 추출하기 위한 trienzyme 방법을 개선하기 위하여 ${\alpha}$-amylase와 folate conjugase 를 동시에 처리하는 방법 (CI)과 기존의 방법대로 protease, ${\alpha}$-amylase, folate conjugase 세가지 효소를 각각 처리하는 방법 (SI)을 비교해 보았으며, 개선된 방법을 이용하여 패스트푸드와 가공식품에 들어있는 엽산 함량을 분석하였다. 1) 12종 88개 식품에 대하여 엽산을 추출하기 위한 기존의 방법 (SI)과 두가지 효소를 동시에 처리하는 CI 방법으로 엽산 함량을 비교한 결과 식품의 종류에 따라 두 방법간에 -6.5%에서 34.0%까지의 차이를 보였다. 식품의 종류별로 paired t-test를 한 결과 9종에서는 유의적 차이를 볼 수 없었으며, 3종에서는 CI 방법이 SI 방법에 비해 유의적으로 높은 것으로 나타났다. 이는 효소 처리시간이 단축됨으로 인해 엽산의 손실이 적었기 때문인 것으로 보이며, 따라서 실험시간을 절약할 수 있는 CI 방법을 활용하는 것이 바람직하겠다. 2) CI 방법으로 식품 중의 엽산을 추출하고 미생물학적 방법으로 엽산 함량을 분석한 결과 햄버거의 100 g당 평균 엽산 함량은 29.0 ${\mu}g$, 치즈버거 35.6 ${\mu}g$, 불고기버거 34.4 ${\mu}g$, 새우버거 33.5 ${\mu}g$, 치킨버거 29.3 ${\mu}g$, 휘시버거 31.7 ${\mu}g$이었다. 비스킷의 엽산 함량은 28.1 ${\mu}g$, 감자튀김은 49.1 ${\mu}g$이었고, 피자 29.7 ${\mu}g$, 샌드위치 31.6 ${\mu}g$, 삼각김밥 24.3 ${\mu}g$으로 나타났으며, 이 값을 1회 분량으로 환산하면 버거종류는 43.1${\sim}$62.0 ${\mu}g$, 감자튀김 53.3 ${\mu}g$, 피자 28.4 ${\mu}g$, 샌드위치 47.4 ${\mu}g$, 삼각김밥 25.7 ${\mu}g$이었다. 3) 가공식품 26종의 엽산함량을 측정한 결과 100 g 당 엽산 함량이 가장 높은 식품은 캔옥수수로 89.5 ${\mu}g$이었고 다음으로는 냉동만두가 80.8 ${\mu}g$이었다. 그 외의 가공식품으로 소시지는 19.6 ${\mu}g$, 비엔나소시지 11.7 ${\mu}g$, 햄 11.5 ${\mu}g$, 튀긴어묵 10.7 ${\mu}g$ 등으로 측정되어 가공식품의 엽산 함량은 대체로 낮은 수준이었으며, 음료의 엽산 함량은 100 g당 1.0${\sim}$5.2 ${\mu}g$으로 매우 낮았다. 4) 본 연구에서 분석한 패스트푸드와 가공식품, 소스, 음료 등 32종의 엽산 함량을 기존의 식품영양가표와 비교하였을 때 식품영양가표의 엽산값은 분석값과 비슷한 경우도 있었으나 0.04${\sim}$140배 까지 차이가 매우 큰 경우도 있었다. 본 연구에서 분석한 식품 중의 엽산 함량은 우리나라 사람들의 엽산 섭취량을 파악하고자 할 때 기초자료로 활용 될 수 있을 것이며, 어린이, 청소년, 젊은 성인층에서 섭취량이 점차 증가하고 있는 패스트푸드와 새롭게 개발되는 가공식품의 엽산 함량을 앞으로도 지속적으로 분석하여 이를 데이터베이스에 반영할 필요가 있다고 여겨진다.

• 한국식품과학회지
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• 제26권5호
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• pp.532-538
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• 1994

Chymotrypsin, trypsin, pancreatin, 그리고 Aspergillus oryzae 유래 단백질분해효소의 in vitro 처리에 의하여 유청단백질(WPI)의 가수분해물(WPH)중 ${\beta}-LG$유래의 항원성변화를 조사하기 위하여 토끼 항${\beta}-LG$항혈청을 이용한 competitive inhibition ELISA(cELISA)와 heterologous PCA를 실시하였다. cELISA에 의하여 WPH의 monovalent항원성을 분석한 결과, 전체적으로 ${\beta}-LG$유래의 monovalent항원성은 효소처리에 의하여 $10^{-1.7}{\sim}10^{-4.1}$배 또는 그 이하로 저하되었으며, 특히 pepsin전처리후 Asp. oryzae유래의 효소로 가수분해한 경우(OUP)의 항원성은 거의 상실되었다. Guinea pig를 이용한 PCA test에 의하여 ${\beta}-LG$유래의 polyvalent항원성을 분석한 결과, WPH의 항원성은 $1/2{\sim}1/128$ 또는 그 이하로 저하되었다. 특히, WPH중에서 열변성이나 pepsin의 전처리없이 Asp. oryzae유래의 효소로 가수분해한 경우(OUN), 가수분해도(DH)가 그다지 높지 않고 monovalent항원성도 여전히 잔존하였음에도 불구하고($10^{-3.2}$배로 저하) 알레르기의 발증과 밀접한 관련이 있는 polyvalent항원성은 거의 상실되었다. 이는 OUN의 분해효율이 아주 높지는 않으나 ${\beta}-LG$상의 항원결정기가 효과적으로 파괴되어, polyvalent항원성이 제거되었기 때문으로 추측된다. 이 결과는, Asp. oryzae유래의 효소를 WPI에 처리하면 우유 알레르기의 주요 원인물질인 ${\beta}-LG$의 polyvalent항원성이 제거됨으로써 저알레르기성 infant formula용 WPH가 제조될 수 있음을 시사하고 있다.

• 대한한의학회지
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• 제26권2호
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• pp.1-12
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• 2005

Objectives: Malignant gliomas are often treated with cisplatin (cis-diamminedichloroplatinum(II), CDDP) and radiation but results remain unsatisfactory. Since malignant glioma displays moderate resistance to conventional therapy, a new treatment modality is needed to improve the outcome of patients with these tumors. The aim of this study was to investigate the effects of the combined use of Jongjihwan(JJH) and cisplatin(CDDP) on cultured malignant glioma cells, A172. Methodss & Results: The combined use of cisplatin and Jeongjihwan had synergistic effects on Al72 cells during 24 hr-incubation, This treatment resulted in a decrease of cell viability, Which was revealed as apoptosis Characterized by activation of caspase-3 protease as well as cleavage of poly ADP-ribose polymerase (PARP) with change of mitochondria membrane potential transition. The expression of members of the Bcl-2 protein family was modulated during co-treatment with Jeongjihwan and cisplatin. Activation of caspase-3 and mitochondrial alterations were central to co-treatment with Jeongjihwan and cisplatin-induced apoptosis. Conclusions: We conclude that co-treatment with Jeongjihwan and cisplatin-induced activation of the mitochondrial pathway enables cell death. Also, we suggest the combined theory of JJH and cisplatin could be a useful method for glioblastoma.

• 한국식품과학회지
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• 제20권3호
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• pp.426-432
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• 1988

${\alpha}-galactosidase$ 및 protease활 을 가지는 효소제 제를 두유(豆乳) 제조시에 처리하여 장내(腸內) 가스발생인자(flatulence factor)를 제거 하는 동시에 두유의 수율(收率)과 소화율(消化率)을 향상시키고자 실험한 결과는 다음과 같다. 두유의 생산량 및 단백질 수율은 효소제(酵素劑) 처리에 의하여 증가하지 않았다. 두유 중 고형분의 양은 pH 6에서 pH10으로 높아질수록, 효소제 처리온도가 $30^{\circ}C$에서 $60^{\circ}C$로 높아질수록 증가하였다. 두유중 TCA가용성 질소화합물의 비율은 효소제의 농도가 높을 수록, pH 4에서 pH10으로 증가 할수록, 처리시간이 길어질수록 증가하였고 처리온도 $50^{\circ}C$까지 증가하는 경향을 보였다. 장내 가스 발생인자로 알려진 raffinose와 stachyose의 함량도 효소제의 농도가 증가할수록, 처리 온도가 $30^{\circ}C$에서 $60^{\circ}C$로 증가함에 따라 많이 감소하였으며 최적 pH는 4-5사이였다.