• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.82-83
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• 2000

수산 생물의 종에 따른 생물학적 다양성은 각기 독특한 성질을 갖는 다양한 효소들이 분포한다는 것을 의미하기도 한다. 이러한 이유로 다양한 효소자원으로서 수산동물의 가공 부산물 또는 폐기물로부터 소화 관련 효소(digestive enzymes)의 회수와 산업적 이용의 잠재성은 매우 크다고 할 수 있다. 수산동물의 소화 관련 효소는 pepsin(Gildberg et al., 1990;1991), chymotrypsin, trypsin(Heu et al, 1995), gastricsin(Sanchea- Chiang and Ponce, 1981) 그리고 elastase(Bjarnason et al., 1993)을 상으로 연구가 이루어지고 있다. 특히 수산동물과 같이 저온에 적응한 냉혈동물의 단백질분해효소는 육상의 온혈동물의 동종 효소보다 낮은 최적온도와 온도안정성 때문에 식품산업에 응용성이 넓다고 하겠다. 따라서 우리나라의 다양한 수산자원의 폐기물 및 가공부산물로부터 소화 관련 효소를 분리하여 식품산업에 있어서 가공보조제(processing aids)로서 이용 가능하리라 생각된다. (중략)

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.4
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• pp.608-616
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• 1996

To propose the use of skipjack processing by-product(SPB) as a food material, the optimal condition for the production of the SPB hydrolysate through enzyme treatment was obtained using RSM(Response Surface Methodology). Among eight pretenses test, Pretense $P^{TM}$ was screened primarily on the aspect of production cost and taste of the product. The extent of autolysis accompanied by endogenous enzyme in the SPB was almost negligible as compared with that of Protense $P^{TM}$ treatment. The derived model equation was within the satisfiable range as indicated by coefficient of $determination(R^2=0.9460)$ and lack of fit(p>0.1) values. From the results of RSM and ridge analysis, the conditions favoring the higllest degree of hydrolysis were: PH 7.2, $51^{\circ}C,$ reaction time of 3.94 hr, substrate concentration of 33.3%, and enzym $e_strate ratio of 0.48%.48%.8%.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.1
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• pp.194-199
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• 2009

An efficient production method of food-grade heamatococcus extract was developed based on stepwise enzymatic hydrolysis. In the first step, Haematococcus pluvialis cells hydrolysis carried out with commercially available exopeptidase(Flavourzyme) and endopeptidase (Alcalase), resulted in increased astaxanthin content. In the second step, proteolytic hydrolyzed H. pluvialis cells treated with hetero-polysaccharides hydrolytic enzyme (Viscozyme). By two-stage treatments using Alcalase and Flavourzyme and Viscozyme, the highest astaxanthin content was obtained. The astaxanthin content was remarkably enhanced by 320% $(529{\mu}g/g\rightarrow2,256{\mu}g/g)$ than that of the non-treated extract. And then, antioxidative activities determined by DPPH method were increased with increasing the astaxanthin content in haematococcus extract prepared by enzymatic hydrolysis.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.73-77
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• 1998

The effects of soymilk residues solubilization by cellulase, protease, koji and yeast were investigated on dry matter and protein yields, amino acid and organic acid contents. Co-treatment of soymilk residues by cellulase and protease gave high dry matter yield and protein yield. Koji treatment followed by yeast fermentation was effective for increasing organic acid content and producing soy sauce-like taste and odor. Organic acid content of fermented hydrolysates was improved by cellulase treatment. Protease treatment rather than koji treatment gave high amino acid content, and cellulase treatment seemed to have little effect on increasing free amino acid content. In sensory evaluation, koji-treated hydrolysate showed higher overall acceptance than other hydrolysates, however it showed lower overall acceptance than commercial fermented soy sauce.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.139-142
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• 1997

Cleaving host immunoglobulins is a well known mechanism of evading host immune reactions exploited by helminth parasites, Secreted cysteine proteases of helminth are a part of enzymes cleaving host IgG. Porogonimw westemani produces at least six different species of the cysteine protease in its developmental stages. This study was undertaken to evaluate comparatively the activities against human IgG by the different enzymes. When the metacercariae, which secrete 27 and 28 kDa cysteine proteases, were incubated in human IgG solution, IgG was degraded at its hinge region. Further incubation resulted complete hydrolysis. From 4-week and 7-week old juveniles and 16-week old adults of p. westemani, five different enzymes at 15, 17. 27 28 and 53 kDa have been purified, if the enzyme with the same molecular mass is regarded to be identical. In cleaving human IgG, each cysteine protease exhibited decreasing activities with age.

• Korean Journal of Microbiology
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• v.28 no.1
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• pp.65-70
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• 1990

The objective of the current study is to elucidate the biological roles of proteinase inhibitor in microorganisms. As the first step, a strain of Streptomyces fradiae was selected as a producer of extracellular proteinase inhibitor. The proteinase inhibitor was purified from culture broth through ultrafiltration, gel-filtration and ion-exchange chromatography. Molecular weight of the proteinase inhibitor was estimated to be 16, 800 by SDS polyacrylamide gel electrophoresis. It was found that the proteinase inhibitor inhibited only alkaline serine proteinases such as subtilisin, $\alpha$-chymotrypsin and Promase E but not trypsin and other proteinases. The mode of inhibition against Pronase E with succinyl-phenylalanine-p-nitroanilide as a substrate was competitive.

• Journal of Life Science
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• v.29 no.10
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• pp.1126-1135
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• 2019

From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.

• Journal of Life Science
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• v.25 no.4
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• pp.385-389
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• 2015

In this study, it is reported that a large polyprotein can be stoichiometrically cleaved by the use of caspase-3-dependent proteolysis. Previously, it has been shown that the proteolytic IETD motif was partially processed when treated with caspase-3, while the DEVD motif was completely cleaved. The cleavage efficiency of the DEVD-based substrate was approximately 2.0 times higher than that of the IETD substrate, in response to caspase-3. Based on this, 3 protein genes of interest were genetically linked to each other by adding two proteolytic cleavage sequences, DEVD and IETD, for caspase-3. Particularly, glutathione-S transferase (GST), maltose binding protein (MBP), and red fluorescent protein (RFP) were chosen as model proteins due to the variation in their size. The expressed polyprotein was purified by immobilized metal ion affinity chromatography (IMAC) via a hexa-histidine tag at the C-terminal end, showing 93 kDa of a chimeric GST:MBP:RFP fusion protein. In response to caspase-3, cleavage products, such as MBP:RFP (68 kDa), MBP (42 kDa), RFP (26 kDa), and GST (25 kDa), were separated from a large precursor GST:MBP:RFP (93 kDa) via SDS-PAGE. The results obtained from this study indicate that a multi-protein can be stoichiometrically produced from a large poly-protein by using proteolytic recognition motifs, such as DEVD and IETD tetra-peptides, for caspase-3.

• Food Engineering Progress
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• v.13 no.2
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• pp.85-91
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• 2009

High hydrostatic pressure and enzymatic hydrolysis (HPEH) was applied to anchovy in order to produce a natural seasoning ingredient. Total soluble solid, amino nitrogen, total nitrogen and the degree of hydrolysis of anchovy hydrolysates were investigated depending on the process parameters such as temperature, pressure, enzyme concentration and enzyme type. The optimal condition for anchovy hydrolysis was confirmed as temperature 50$^{\circ}C$, reaction time 24 hrs, pressure 50 MPa and enzyme concentration 0.6% in HPEH treatment. HPEH treatment showed more effective in overall properties of anchovy hydrolysis than those of control. All anchovy hydrolysates produced by HPEH treatment were increased more 1.5-2.6 times of total free amino acid than that of control. From these results, the HPEH treatment appears to be an effective and economic process to produce a natural seasoning ingredients.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.237-244
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• 1989

A soluble intracellular protease from cells of Pseudomonas carboxydovorans, a carboxydobacterium, grown on nutrient broth was purified 68-fold in five steps to better than 95% homogeneity with a yield of 2.4% using azocasein as a substrate. The enzyme activity was not detected from cells grown on pyruvate, succinate, acetate, or CO as a sole source of carbon and energy. The molecular weight of the native enzyme was determined to be 53,000. Sodium dodecyl sulfate-gel electrophoresis revealed the purified enzyme a monomer. The enzyme was found to be a serine-type protease. The enzyme activity was inhibited completely by several divalent cations such as $Cd^{2+}, Cu^{2+}, Hg^{2+}$, and $Fe^{2+}$. The enzyme was also inhibited by EGTA, but was stimulated by iodoacetamide. The optimal pH and temperature for the enzyme reaction were found to be 8.0 and $50^{\circ}C$, respectively. The enzyme was inactive on CO dehydrogenase.