• 대한예방한의학회지
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• 제16권2호
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• pp.95-111
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• 2012

Objective : The water extract of Danguiyonghoihwan (DGYHW) has been traditionally used in treatment of tinnitus in Oriental Medicine. However, little is known about the mechanism by which DGYHW rescues auditory neuronal cells from injury damages. Therefore, in this study I effort to elucidate the mechanism of the cytoprotective effect of the DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Methods : I determined the elevated cell viability by DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Glutamate induced neuronal damage in oranotypic explant culture also, glutamate decreased cell viability on VOT-33 cells but pretreatment with DGYHW inhibited cell death. Results : One of the main mediator of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pretreatment with DGYHW inhibited this ROS generation from glutamated-stimulated VOT-33 cells. Also, I identified that the ROS-induced DCF-DA green fluorescence is reduced by DGYHW pretreatment. The critical markers of apoptotic cell death were cleavages of procaspase-3 protease protein. So I checked the expression level and cleavage of procaspase-3 protease protein. Glutamate-treated VOT-33 cells were shown to have cleavage of procaspase-3 protease proteins and following reduction of expression of these proteins. But I found that pre-treatment with DGYHW protects glutamate-induced changes of biochemical marker protein, caspase-3. Conclusion : These findings indicated that DGYHW may prevent cell death from glutamate induced VOT-33 cell death by inhibiting the ROS generation and modulation of protein expressions in procaspase-3, catalase and Bcl-2.

• 한국식품영양학회지
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• 제15권4호
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• pp.331-336
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• 2002

본 연구는 두유에 칼슘을 강화하고자 두유단백질의 칼슘내인성을 높이기 위한 방법으로 단백분해효소를 처리한 경우 효소처리 전, 후에 나타나는 단백질의 구조적 특성을 비교 분석하였다. 표면소수도는 SMP

• 한국식품과학회지
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• 제24권1호
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• pp.101-106
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• 1992

제주도 전통주인 좁쌀약주를 청징화시키기 위하여 원심분리에 의한 청징화 방법, 단백효소제(papain, bromelain, ficin)에 의한 청징화 방법과 한외여과에 의한 청징화 방법들을 검토하여 좁쌀청주의 가장 효과적인 청징화 방법을 모색하고자 하였다. 원심분리에 의해서만은 혼탁물질의 완전한 제거는 불가능하였으며, 단백효소제를 이용한 경우에서는 효소농도 $5{\times}10^{-4}%(w/v)$ 작용시간 2시간, 작용온도 $35^{\circ}C$에서 매우 우수한 청징효과를 나타냈다. 효소별 청징도는 papain과 bromelain은 거의 비슷한 효과를 보였으나, ficin의 청칭력은 약간 떨어졌다. 한외여과(여과막 크기 100k와 $0.22{\mu}m$막)에 의한 청징화는 효소에 의한 최고의 청징화 수준보다 약간 우수한 청징효과를 보였으며, 효소처리에 비하여 향미성분의 손실이나 색도가 좀더 나아서 UF에 의한 청징화 방법이 간단하고 경제적인 방법이었다.

• Journal of Microbiology and Biotechnology
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• 제27권11호
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• pp.1952-1960
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• 2017

Recently, soybean isoflavone aglycones (i.e., daidzein and genistein) and ${\gamma}-aminobutyric$ acid (GABA) have begun to receive considerable consumer attention owing to their potential as nutraceuticals. To produce these ingredients, multiple microorganisms and their enzymes are commonly used for catalysis in the nutraceutical industry. In this work, we introduce a novel fermentation process that uses whole-cell biocatalysis to accelerate GABA and isoflavone aglycone production in doenjang (a traditional Korean soybean paste). Microbial enzymes transform soybean isoflavone glycosides (i.e., daidzin and genistin) and monosodium glutamate into soybean isoflavone aglycones and GABA. Lactobacillus brevis GABA 100 and Aspergillus oryzae KACC 40250 significantly reduced the production time with the aid of a protease. The resulting levels of GABA and daidzein were higher, and genistein production resembled the levels in traditional doenjang fermented for over a year. Concentrations of GABA, daidzein, and genistein were measured as 7,162, 60, and $59{\mu}g/g$, respectively on the seventh day of fermentation. Our results demonstrate that the administration of whole-cell L. brevis GABA 100 and A. oryzae KACC 40250 paired with a protease treatment is an effective method to accelerate GABA, daidzein, and genistein production in doenjang.

• 대한의생명과학회지
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• 제16권2호
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• pp.105-112
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• 2010

Methylprednisolone (MPD) is a synthetic glucocorticoid drug used in treatment of many neurological diseases and neurotraumas, including spinal cord injuries. Little is known of the mechanism of MPD in neuronal cells, particularly the genetic expression aspect. DD-PCR was used in identification of genes expressed during MPD treatment of PC12 cells. We have isolated 3 predicted up- or down-regulated genes, which are differentially expressed in neurons by MPD. One of these genes, USP16 (ubiquitin specific protease 16), is the deubiquitinating enzyme that is up-regulated by MPD in neurons. In order to observe the effect of MPD on USP16 gene expression, PC12 cells were treated under several experimental conditions, including endoplasmic reticulum stress drugs. We have isolated the total RNAs in PC12 cells and detected USP16 and ER related genes by RT-PCR. Because its expression pattern is similar to expression of ER chaperons, USP16 gene expression is strongly associated with unfolded protein response. A meaningful negative effect on each tissue treated by methylprednisolone is not shown in vivo. USP16 gene expression is suppressed by LY294002 (phosphatidylinositol 3-kinase inhibitor), which suggests that USP16 gene expression is regulated by the phosphatidylinositol 3-kinase pathway.

• 한국가금학회지
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• 제34권1호
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• pp.9-14
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• 2007

본 연구는 산란계 사료내 갯벌 미생물 유래 단백질 분해 효소제를 첨가하였을 때 계란 품질, 영양소 소화율 및 혈청 내 총 단백질 함량에 미치는 영향을 알아보기 위해 실시하였다. 사양 시험은 29주령 ISA brown 갈색계 252수를 공시하였고, 사양 시험은 8주간 실시하였다. 옥수수-대두박 위주의 기초 사료구(CON; basal diet), 단백질 분해 효소제 0.05, 0.10% 첨가구(PRO1; PRO2)로 처리구당 7반복, 반복당 12수씩 완전임의 배치하였다. 전체 사양 시험 기간 동안, 산란율은 처리구간에 유의적인 차이를 보이지 않았다(P>0.05). 난황고의 변화량은 PRO1 처리구가 CON 처리구와 비교하여 유의적으로 높게 나타났다(P<0.05). 난중의 변화량은 PRO2 처리구가 CON과 PRO1 처리구와 비교하여 유의적으로 높게 나타났다(P<0.05). 난각 품질, 난황색, haugh unit 및 난황 계수에서는 처리구간에 유의적인 차이를 보이지 않았다(P>0.05). 건물 소화율에서는 CON과 PRO2 처리구가 PRO1 처리구와 비교하여 유의적으로 높게 나타났고(P<0.05), 질소 소화율에서는 PRO2 처리구가 CON 처리구와 비교하여 유의적으로 높게 나타났다(P<0.05). 혈청내 총 단백질 함량에서는 처리구간에 유의적인 차이를 보이지 않았다(P>0.05). 결론적으로, 산란계 사료내 갯벌 미생물 유래 단백질 분해 효소제의 첨가는 난황고, 난중 및 영양소 소화율을 개선시켰다.

• 동의생리병리학회지
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• 제18권3호
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• pp.774-782
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• 2004

The water extract of Dancheonhwan (DCH) has been used to treat ischemic brain and heart damage in oriental medicine. However, little is known about the mechanism by which the water extract of DCH rescues cells from ischemic damage. Therefore, this study was designed to investigate the protective mechanisms of DCH on the H₂O₂-induced toxicity in H9c2 cardiomyoblast cells. Treatment of H₂O₂ markedly decreased the viability of H9c2 cardiomyoblast in a dose-dependent and time-dependent manner. The nature of H₂O₂-induced toxicity of H9c2 cells resulted from apoptotic death confirmed with genomic DNA fragmentation. DCH increased the viability of H₂O₂-treated H9c2 cells by about 23%, and partially suppressed the genomic DNA fragmentation and PARP cleavage. H₂O₂ also activated caspase-3 protease and -9 protease, but not both caspase-6 protease and -8 protease. H₂O₂ induced the mitochondria dysfunction, including mitochondria membrane permeability transition (MPT) and cytosolic release of cytochrome c from mitochondria, which was prevented in part by pretreatment of DCH. N-acetylcystein (NAC), a free-radical scavenger, alone increased the viability of H₂O₂-treated H9c2 cells in a dose-dependent manner. Furthermore, the combination of NAC with DCH significantly increased the viability of the H₂O₂-treated H9c2 cells in a dose-dependent manner. These data indicate that DCH has the protective effect on ROS-induced apoptosis of cadiomyoblast H9c2 cells.

• Bulletin of the Korean Chemical Society
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• 제33권9호
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• pp.2883-2889
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• 2012

LL-37 is the only antimicrobial peptide (AMP) of the human cathelicidin family. In addition to potent antimicrobial activity, LL-37 is known to have the potential to inhibit lipolysaccharide (LPS)-induced endotoxic effects. To provide the stability to proteolytic digestion and increase prokaryotic selectivity and/or anti-endotoxic activity of two Lys/Trp-substituted 19-meric antimicrobial peptides (a4-W1 and a4-W2) designed from IG-19 (residues 13-31 of LL-37), we synthesized the diastereomeric peptides (a4-W1-D and a4-W2-D) with D-amino acid substitution at positions 3, 7, 10, 13 and 17 of a4-W1 and a4-W2, respectively and the enantiomeric peptides (a4-W1-E and a4-W2-E) composed D-amino acids. The diastereomeric peptides exhibited the best prokaryotic selectivity and effective protease stability, but no or less anti-endotoxic activity. In contrast, the enantiomeric peptides had not only prokaryotic selectivity and anti-endotoxic activity but also protease stability. Our results suggest that the hydrophobicity and ${\alpha}$-helicity of the peptide is important for anti-endotoxic activity. In particular, the enantiomeric peptides showed potent anti-endotoxic and LPS-neutralizing activities comparable to that of LL-37. Taken together, both a4-W1-E and a4-W2-E holds promise as a template for the development of peptide antibiotics for the treatment of endotoxic shock and sepsis.

• Applied Biological Chemistry
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• 제46권3호
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• pp.176-182
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• 2003

한국재래간장으로부터 혈전용해효소를 강하게 생산하는 균주를 선발하고 이를 Bacillus subtilis K7로 동정하였다. B. subtilis K7이 생산하는 혈전용해성 protease를 정제하여 분자량을 확인한 결과 21,500 Da이었다. 정제된 효소의 최적반응조건은 $40^{\circ}C$와 pH 9.0이었으며 pH 5.0라서 12.0까지 안정하고 $50^{\circ}C$에서 20분간 열처리한 후에도 50%이상의 효소활성을 가지며 EDTA, CDTA 및 iodoacetat에 실활하는 효소이었다. 이 효소의 fibrin에 대한 Km 값은 $1.8{\times}10^{-2}$ M이었다.

• Journal of Animal Science and Technology
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• 제62권3호
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• pp.313-320
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• 2020

This research was performed to investigate the hypothesis that dietary mono-component protease (PRO) might improve growth performance, nutrient digestibility, and carcass characteristics of growing-finishing pigs. A total of eighty-four pigs [Duroc × (Landrace × Yorkshire), 25.3 ± 2.16 kg initial body weight] were randomly assigned to three dietary treatments (7 replicates/treatment; 2 barrows and 2 gilts/replicate) in a randomized complete block design (block = sex). The dietary treatments were prepared as follows; (1) a positive control (PC) as a typical growing-finishing diet based on corn and soybean meal, (2) PC added with 0.015% of PRO (PCPRO), and (3) a negative control (NC) added with 0.015% of PRO (NCPRO). The NC had a lower concentration of crude protein (CP) compared with PC. The PRO was a commercial product that contained 75,000 protease units/g and derived from Nocardiopsis prasina produced in Bacillus licheniformis. Dietary treatments were offered to pigs during growing and finishing periods. Measurements were growth performance, apparent total tract digestibility (ATTD) of nutrients, and carcass characteristics. The PCPRO and/or NCPRO increased average daily gain (ADG) and gain to feed ratio (G:F) during growing (p < 0.10), finishing (p < 0.05), and growing-finishing periods (p < 0.10) compared with PC. Furthermore, pigs fed PCPRO and NCPRO had higher (p < 0.05) ATTD of CP and energy during growing and/or finishing periods than those fed PC. In conclusion, the supplementation of PRO in diets improved growth performance and protein digestibility of growing-finishing pigs.