• Journal of Microbiology
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• 제33권3호
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• pp.244-251
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• 1995

An extracellular protease of Salmonella schottmulleri was purified from culture filtrate by using 0-75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, Ultrogel HA chromatography and Sephacryl S-200 HR molecular sieve chromatography. To measure enzyme activity, synthetic dipeptide substrate (CBZ-arg-arg-AFC) with low molecular weight was employed as substrate. The molecular weight of the purified enzyme was approximately 80 kDa when determined by gel filtration on Sephacryl S-200 HR and 73 kDa when estimated by SDS-PAGE. The isoelectric point was 5.45. The activity of the purified enzyme was inhibited by metal chelating agesnts such as EDTA and 1.10-phenanthroline. The divalent cations, such as Ca$\^$2+/, Zn$\^$2+/, Fe$\^$2+/, Mg$\^$2+/ enhanced its activity. These results suggested that it was a metalloprotease. It had a narrow pH optimum of 6.5-7.5 with a maximum at pH 7.0 and a temperature optimum of 40.deg.C. It was stable at least for 1 week at 40.deg.C and maintained its activity for 24 hours at 50.deg.C, but it was rapidly inactivated at 65.deg.C. This protease was shown to be sensitive to sodium 50.deg.C, but it was rapidly inactivated at 65.deg.C. This protease was shown to be sensitive to sodium 50.deg.C, but it was rapidly inactivated at 65.deg.C. This protease was shown to be sensitive to sodium 50.deg.C, but it was rapidly inactivated at 65.deg.C. This protease was shown to be sensitive to sodium dodecyl sulfate (SDS) and was inactivated in a dose-dependent manner. However, it was resistant to Triton X-100 and the activity was enhanced to 32.3% with treatment of 0.025% Triton X-100.

• Applied Biological Chemistry
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• 제38권6호
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• pp.554-562
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• 1995

밥의 식미가 다른 추청벼(자포니카형), 조생통일벼(통일계), IR 36(인디카형)의 세 품종의 쌀을 시료로 하여 단백질 사수분해 효소와 이황화 결합 환원제 처리가 쌀의 이화학적 성질, 호화 특성 및 가용성 탄수화물용출 양상에 미치는 영향에 관하여 실험하였다. 세 품종 쌀의 아밀로오스 함량은 IR 36이 24.56%로 가장 높고 조생통일벼가 20.63%, 추청벼가 20.36%였다. 팽윤력과 용해도는 IR 36이 특히 낮았다. 시차 주사열량계로 조사한 호화 엔탈피와 X-선 회절에 의한 결정성은 IR 36이 가장 컸다. 쌀가루의 단백질을 효소로 가수분해 시켰을 때 가열 중 수분 흡수량이 증가하였고, 쌀에 2-mercaptoethanol 처리를 하였을 때 세 품종 모두 수분흡수 속도가 커지고 수분 흡수량이 증가하였으며 특히 IR 36에서 현저한 증가를 보였다. 아밀로그램의 호화개시온도는 추청벼가 가장 낮고 IR 36이 높았으며 $50^{\circ}C$의 냉각 점도와 setback은 IR 36이 가장 컸다. 효소처리와 2-mercaptoethanol 처리는 아밀로그램의 점도 감소를 초래하여 쌀알 중의 단백질이 전분립의 팽윤을 제한하고 전분립에 강도를 부여하는 것을 알 수 있었다. 효소처리를 한 팔 전분 현탁액에서도 쌀가루만큼 뚜렷하지는 않았으나 아밀로그램의 점도가 감소하였다. 가용성 탄수화물의 분자량 분포는 세 품종간에 차이가 있었다. 추청벼가 용출된 가용성 탄수화물의 양이 가장 많았고, 큰 분자량의 분율이 다른 품종보다 높았다. 단백질 가수분해 효소처리는 쌀가루의 가용성 탄수화물 용출량, 특히 큰 분자량($K_{av}\;0.3$ 이상)의 용출을 증가시켰다. 이 증가량은 IR 36에서 가장 컸고 조생통일벼, 추청벼의 순이었다.

• 복식문화연구
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• 제12권5호
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• pp.737-742
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• 2004

The purpose of this study is restoration for tearing strength of the durable press (DP) finished 100% cotton fabric by enzymatic treatment. Dimethylol Dihydroxy Ethylene Urea (DMDHEU) was used as a DP finish chemical. Enzymes (cellulase, pectinase, protease, lipolase) were selected based on their specific reaction activities. Ideal application of the enzymes for this work was to remove cross-links created by DMDHEU on the surface of the fibers to offer migration property between microstructures of cellulose, yet cross-links that exist inside of the fibers are still remained to impart effect of wrinkle resistance. Physical characteristics (tearing strength, wrinkle recovery, FT-IR) of enzyme treated samples were measured and compared. It was found out that, in case of enzyme treatment, most of enzymes didn't have a great effect on tearing strength, but, in case of Protease, tearing strength increased at DMDHEU 2% treatment. As a result of an experiment on wrinkle recovery of the textiles treated with enzyme making density of DMDHEU different whenever respective experiment was made, it was discovered that density of DMDHEU increased as wrinkle recovery increased and, in the relation to enzyme treatment especially in Lipase enzyme treatment, the lesser density of DMDHEU, the more wrinkle recovery increased.

• 한국미생물·생명공학회지
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• 제21권6호
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• pp.513-519
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• 1993

The alkaline protease in the polyhedra preparation of Spodoptera litura nuclear polyhedrosis virus was successfully inactivated by heating at 100C for 20 minutes. SDS-PAGE analysis indicated that heat inactivated polyhedra is composed of major proteins of 31kDa and presumptive its polymer protein of 62kDa. However, this polyhedra was converted into several smaller molecular weight proteins when treated with midgut juice, but not by treatment with heat-inactivated midgut juice.

• 한국식품영양과학회지
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• 제27권6호
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• pp.1077-1085
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• 1998

The effects of yeast on the fermentation of kimchi were investigated. The treatments were divided into two groups; yeast treatment during salting of Chinese cabbage(YS) and yeast treatment added in kimchi preparation(YF kimchi). The edible periods of the kimchi after yeast treatment during salting (YS kimchi) was extended 4~5 days by the results of pH, acidity, sensory quality. The activities of amylase, polygalacturonase and galactosidase of YS kimchi were retained at low levels compared to non treated condition throughout all fermentation periods, whereas protease activity was not significant different from the non treated condition. In addition, the contents of total hexose and uronic acid did not show remarkable change throughout fermentation, but total pentose was decreased by more than 7% at the early middle stage of fermentation(7~14 day after soaking). The change of free amino acid content was decreased by 16~44% than the non treated condition. In contrast, in the YF kimchi, the sensory quality was not good. The activities of amylase, protease, polygalacturonase and gal actosidase were appreciably higher than that of the non treated condition. Meanwhile, the contents of total hexose, total pentose and uronic acid, as products of degradation of cell wall constituents by the above enzymes, were decreased by 18~68% throughout fermentation than the non treated con dition, and total free amino acids were higher than the YS kimchi. Thus, yeast treatment during salting was found to be more effective to extend the edible periods of the kimchi.

• Journal of Nutrition and Health
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• 제51권6호
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• pp.599-606
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• 2018

Purpose: Various plants, herbal medicines, and marine foodstuffs have been used in kimchi preparation to improve its overall quality. Teff, which is rich in minerals and starches, facilitates stable blood glucose levels and is well-suited for use in gluten-free products; hence, it can be used to reinforce the mineral composition of kimchi. In this study, we probed the antioxidant activities of hydrolysates prepared by treatment of brown teff with three proteases under different conditions. Methods: The mineral composition of brown teff was determined by inductively coupled plasma spectrophotometry-mass spectrometry, and we established optimal hydrolysis conditions by determining the total phenol and flavonoid contents of teff hydrolysates obtained using three different proteases (protamax, flavourzyme, and alcalase), two different protease concentrations (1 and 3 wt%), and three different incubation times (1, 2, and 4 h). The antioxidant activity of the hydrolysates was further investigated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, total antioxidant capacity (TAC), and ferrous reducing antioxidant power (FRAP) assays. Results: Brown teff was rich in I, K, Mg, and Ca, and the highest total phenol content ($24.16{\mu}g/mL$), total flavonoid content ($69.08{\mu}g/mL$), and TAC were obtained for 1 wt% protamax treatment. However, the highest DPPH scavenging activity and FRAP values were observed for hydrolysates produced by alcalase and flavourzyme treatments, respectively. Conclusion: Treatment of brown teff with proteases affords hydrolysates with significantly increased antioxidant activities and high total phenol and flavonoid contents, and these antioxidant activities of teff hydrolysates have the potential to enhance the quality and functionality of kimchi in future applications.

• 생명과학회지
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• 제16권4호
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• pp.598-604
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• 2006

$Ca^{2+}$-농도 의존적으로 활성화되는 neutral protease calpain에 의한 단백질 분해는 세포의 성장을 조절하는데 중요한 단백질들의 역할에 매우 중요한 역할을 한다. Cyclin의 분해는 세포주기의 진행을 위한 필연적인 과정이다. D-type cyclins는 외부자극이나 신호에 의하여 세포주기의 G1 초기에 합성이 된 후 cyclin-dependent kinases (cdk4 및 cdk6)와의 결합하여 세포주기 S기 진입을 촉진하는 역할을 한다. 본 연구에서는 MDA-MB-231 인체 유방암세포에서 cyclin D3 단백질이 calpain protease에 의하여 전사 후 수준에서 조절 받고 있음을 제시하였다. 본 실험의 조건에서 lovastatin과 actinomycin D가 처리된 MDA-MB-231 세포에서 cyclin D3 단백질의 발현이 완전히 사라졌지만, calpain inhibitor인 LLnL의 처리에 의하여 정상 수준으로 회복되었음을 알 수 있었다. 그러나 26S proteasome의 선택적 억제제인 lactacystin, the lysosome 억제제인 ammonium chloride 및 chloroquine, serine protease 억제제인 PMSF는 동일 조건에서 lovastatin과 actinomycin D 처리에 의한 cyclin D3의 발현저하를 억제하지는 못하였다. In vitro 조건에서 순수 분리된 calpain은 cyclin D3 단백질을 $Ca^{2+}$ 농도 의존적으로 분해하였으며, cyclin D3 단백질의 half-life는 LLnL 처리에 의하여 매우 유의적으로 증가되었다. 이러한 결과는 cyclin D3 단백질이 $Ca^{2+}$에 의해 활성화 되는 protease calpain에 의해 조절됨을 보여준다.

• Toxicological Research
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• 제19권3호
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• pp.197-203
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• 2003

Apicidin, a histone-deacetylase inhibitor, has been successfully used to inhibit the growth of cancer cells. In this study, the apoptotic potential and mechanistic insights of apicidin were investigated in human myeloid leukemia U937 cells. Treatment of U937 cells with apicidin resulted in a decrease of cell viability with apoptotic characteristics, including chromatin condensation and ladder-pattern fragmentation of genomic DNA. Apicidin converted the procaspase-3 protease to catalytically active effector protease, resulting in subsequent cleavage of poly (ADP-ribose) polymerase (PARP) and inhibitor of caspase-activated deoxyribonuclease (ICAD). In addition, apicidin induced the activation of caspase-9 protease and the cytosolic release of mitochondrial cytochrome c with mitochon-drial membrane potential transition. Moreover, apicidin transiently increased the expression of Fas and Fas ligand proteins. Taken together, the results suggest that apicidin induces apoptosis of U937 cells through activation of intrinsic caspase cascades and Fas/FasL system with mitochondrial dysfunction.

• 한국식품영양과학회지
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• 제24권3호
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• pp.427-433
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• 1995

The optimal condition for hydrolysis of oyster was evaluated with proteases using response surface methodology(RSM). Among 11 commerical proteases, APLTM 440 was selected as the suitable protease for producing oyster hydrolysate on the basis of cost per unit enzyme activity. The effect of autolysis on degree of hydrolysis in oyster was negligible comparing to that of APL 440 protease treatment. From RSM and ridge analysis, the conditions favoring the highest degree of hydrolysis were pH 9.95, 61.1$^{\circ}C$, 2.64 hr reaction time, 49.2% substrate, and 0.35% enzyme/substrate ratio. Oyster hydrolysate prepared under optimal conditions shwoed virtually 51.98% of hydrolysis.

• BMB Reports
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• 제33권2호
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• pp.112-119
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• 2000

Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.