• Title/Summary/Keyword: 기질단백분해효소

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Comparison of the Influence of Carbon Substrates on the Fibrolytic Activities of Neocallimastix sp. NLRI-3 (탄소원의 종류가 반추위 혐기 곰팡이 Neocallimastix sp. NLRI-3의 섬유소 분해효소 활력에 미치는 영향 비교)

  • 손호진;송재용;최낙진;하종규;장종수
    • Journal of Animal Science and Technology
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    • v.48 no.3
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    • pp.415-424
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    • 2006
  • The purpose of this study was to investigate the fungal growth and enzyme production under different carbohydrate substrate conditions. The anaerobic fungus Neocallimastix sp. NLRI-3 isolated from the rumen of Korean native goat was incubated with different carbohydrate media containing 0.2% of glucose, starch, rice straw, filter paper, carboxymethyl cellulose(CMC), Sigmacell cellulose, xylan or xylose, respectively. The culture head gas production was the highest in the culture of filter paper medium, and the lowest in the culture of CMC medium at 96h incubation (P<0.05). The fungal zoospore production reached peak at 72h incubation, and its number was the highest in rice straw medium among the treatments (P<0.05). At 96h incubation, carboxymethyl cellulase(CMCase) activity was the highest in the culture of filter paper medium and the lowest in the culture of starch medium (P<0.05). While xylanase activity was the highest in the culture of rice straw medium and the lowest in the culture of xylose medium(P<0.05) at 72h incubation. There were no differences in culture supernatant protein expression among the treatments. However, the patterns of enzyme expression were different among the treatments with zymogram analysis. Six CMCases and 4 xylanase were detected from the results of zymogram analysis. Therefore the present study indicating that the fungal enzyme expression could be stimulated with insoluble substrates in the culture medium.

Expression of Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 after Administration of Endotoxin in Diabetic Rats (내독소로 자극된 당뇨 쥐에서 단백분해효소와 그 억제제 발현)

  • Seo, Ki Hyun;Choi, Jae Sung;Na, Joo Ok;Uh, Soo Taek;Kim, Yong Hoon;Park, Choon Sik
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.3
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    • pp.256-264
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    • 2006
  • Background: An acute lung injury(ALI) is characterized by the recruitment, activation, and apoptosis of inflammatory cells, numerous products released by inflammatory cells such as reactive oxygen species, inflammatory mediators, and a variety of proteolytic enzymes. It was reported that bacterial infections in diabetics showed impaired PMN functions such as reduced PMN respiratory burst and decreased microbicidal activity in inflamed tissue. However, the effect of the proteinase - inhibitor (MMP-9 vs TIMP-1) in ALI in diabetics is unclear. This study evaluated the differences in the expression of MMP-9 and TIMP-1 after the stimulation of endotoxin in a rat model. Methods: Six-week-old male Sprague-Dawley rats were classified into normal, DM, LPS and DM+LPS groups. The peripheral blood, BAL fluids, and lung tissues were obtained from individual rats. The MMP-9 activity was measured by gelatin zymography and the TIMP-1 level was measured by Western blotting. Results: The total BAL cells of the DM-LPS groups were significantly lower than the LPS groups (p < 0.01). The MMP-9 activities in the serum were higher in the DM+LPS groups than in the other groups. The MMP-9 activities in the BAL fluids were significantly higher in the DM+LPS group than in the normal and diabetic rats (p < 0.05). TIMP-1 expressions in the BAL fluids were significantly lower in the DM+LPS group than other groups (p < 0.05). The ratio between MMP-9 and TIMP-1 in the BAL fluids was significantly higher in the DM+LPS groups (p < 0.05). Conclusion: In ALI in diabetics the higher MMP-9 activity and lower TIMP-1 level are believed to prolonged and intensify the course of inflammation.

Proteolytic Properties of Cathepsin L, Chymotrypsin, and Trypsin from the Muscle and Viscera of Anchovy, Engraulis japonica (멸치 육과 내장으로부터 분리한 Cathepsin L, Chymotrypsin 및 Trypsin의 단백질분해 특성)

  • PYEUN Jae-Hyeung;HEU Min-Soo;CHO Deuk-Moon;KIM Hyeung-Rak
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.5
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    • pp.557-568
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    • 1995
  • Proteolytic properties of enzymes from the muscle and viscera of anchovy have been examined. Cathepsin L, chymotrypsin, and trypsin showed similar Km values for casein. However, they had higher Km values for myofibrillar proteins than those for casein. The $k_cat$ of cathepsin L and chymotrypsin for myofibrillar proteins were higher than that of trypsin, and also cathepsin L and chymotrypsin caused higher hydrolysis in myofibrillar proteins of anchovy and yellowtail. In the presence of sodium chloride$(0-25\%)$, proteolytic activity for myofibrillar proteins from yellowtail was higher than that for casein. Proteolytic activity was decreased with the increase of sodium chloride concentration. Cathepsin L had been less affected by NaCl concentration and temperature on the hydrolysis of myofibrillar proteins than chymotrypsin and trypsin. Cathepsin L and chymotrypsin were move responsible to the autolysis of muscle proteins from fish than trypsin.

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Stability and Optimization of Crude Protease Extracted from Korean Kiwifruits (국내산 키위에서 추출한 protease 조효소액의 안정성과 최적화에 관한 연구)

  • Kim, Mi-Hyun;Rho, Jeong-Hae;Song, Hyo-Nam
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.554-558
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    • 2010
  • In the study, the protease activity of kiwifruit (Actinidia deliciosa Planch) cultivated in Korea was estimated, with specific examination of proteolytic effects on myofibrilar protein. The crude protease extract of kiwifruit was prepared in two ways; one in which the kiwifruit was homogenized with buffer followed by centrifugation, and the other were the supernatant was precipitated by saturated ammonium sulfate followed by dialysis. The former had 21.23 mM/mL of protease activity, which corresponded to 112.28 mM/g kiwifruit utilized, and the latter had 11.58 mM/mL and 45.80 mM/g of kiwifruit. The crude protease extract of the kiwifruit showed high specificity for casein substrate followed by bovine serum albumin, egg white, collagen, and elastin, in order. The enzyme lost proteolytic activity in acidic conditions such as pH 2-3, and at high temperatures over $60^{\circ}C$. It showed optimal activity in both pH 3.0 and pH 7.5 as well as at $40^{\circ}C$ for casein substrate and at $50^{\circ}C$ for myofibrilar protein substrate. The proteolytic activity toward casein was high with up to 0.5M salt, followed by a sharp decrease beyond this concentration. On the other hand the proteolytic activity for myofibrilar protein decreased steadily with increasing of salt concentration. Kiwifruit has been used as a for meat tenderizer for in home cooking and these results support the its tenderizing effectiveness of kiwifruit especially for Korean style marinating of meat for cooking.

PZ-peptidase activities in Streptococcus sanguis and other oral bacteria (Streptococcus sanguis와 여타 구강세균이 생산하는 PZ-peptidase 활성)

  • 최선진
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.143-148
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    • 1983
  • The occurrence of PZ-peptidase in Streptococcus sanguis and other oral bacteria was investigated utilizing washed whole cells as the enzyme source and PZ-pentapeptide as its substrate. Under the culture conditions employed in the present study. Streptococcus sanguis strains, fresh isolates as well as laboratory strains, produced a broad range of the enzyme activity (0.5-7.9 unit/mg protein). The strains of both Streptococcus mutans and Lactobacilli showed low levels of activity (0-0.5 unit/mg protein for S. mutans). As compared with the enzyme activities of other bacteria, a moderate range of activity was produced by the strains of Strptococcus mitis nad Strptoccus salivarius. Actinomyces strains, like those of S. sanguis, produced a varying amount of activity (0-9.8 unit/ mg protein). A possible involvement of the oral bacterial PZ-peptidase in the metabolism of human saliva proteins is discussed.

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Hydrolysis of 7S and 11S Soy Proteins by Commercial Proteases (단백분해효소(蛋白分解酵素)에 의한 대두(大豆) 7S 및 11S 단백질(蛋白質)의 가수분해(加水分解))

  • Kang, Yeung-Joo;Lee, Ki-Chun;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.338-343
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    • 1988
  • Selected kinetic parameters and degree of hydrolysis(DH) were measured using commercial proteases(trypsin, alcalase and pronase) to study the affinity of these enzymes to 7S and 11S soy proteins. Electrophoretic patterns of the hydrolysates were also investigated. In general, the order of affinity between the proteins and the proteases was 11S(protein-rich fraction)and 7S PRF for unheated proteins, and 7S PRF and 11S PRF for preheated proteins. Substrate inhibition was present at a substrate concentration of 1.5% or higher when preheated protein was used as the substrate. The maximum DH values of alcalase were obtained from 7S PRF(60%) and 11S PRF(80%) at 1 hr hydrolysis, respectively. Trypsin hydrolyses did not affect 11S soy protein but the acidic subunits in contrast to alcalase and pronase hydrolyses which changed almost all subunits. Alcalase hydrolysis induced distinct changes on 2S soy protein.

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Studies on the Improvements of Functional Properties of Sardine Protein by Plastein Reaction -1. Synthetic Conditions of Plasteins from the Enzymatic Hydrolysate of Sardine Protein- (Plastein반응을 이용한 정어리 단백질의 기능성 개선에 관한 연구 -1. 정어리 분말단백질의 pepsin가수분해물을 이용한 plastein의 합성조건-)

  • Kim, Se-Kwon;Kwak, Dong-Chae;Cho, Duck-Jae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.3
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    • pp.233-241
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    • 1988
  • In order to develop a new type of food source for the effective utilization of fish protein, plastein reaction was applied to improve the functional properties of sardine protein. Conditions necessary for optimal plastein productivity from sardine protein using pepsin, ${\alpha}-chymotrypsin$, protease(from Aspergillus saitoi) and papain were established. Sardine protein concentrate was hydrolyzed with pepsin yielding an approximate degree of hydrolysis of 78.4%. Enzyme induced plastein was optimized at : pH 4 for pepsin, pH 7 for ${\alpha}-chymotrypsin$, pH 5 for pretense and pH 6 for papain : Substrate concentrate 40% for pepsin and ${\alpha}-chymotrypsin$, 50% for pretense and papain : the time of incubation, 24hr : enzyme/substrate ratio, 1 : 100(w/v) incubation temperature, $50^{\circ}C$.

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Development of Natural Meat-like Flavor Based on Maillard Reaction Products (Maillard 반응 생성물을 이용한 천연 육류향의 제조)

  • Moon, Ji-Hye;Choi, In-Wook;Park, Yong-Kon;Kim, Yoon-Sook
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.129-138
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    • 2011
  • Hydrolyzed wheat gluten (HWG) and low glutamic acid (Glu) hydrolyzed wheat gluten with different quantities of NaCl were reacted with several precursors to develop natural meat flavor based on Maillard reaction products (MRP). The MRP based flavors were analyzed for their pH, browning index, DPPH radical scavenging effect, and sensory properties. Synthetic meat flavor from low Glu hydrolyzed wheat gluten with 7% NaCl and ribose, cysteine, methionine, thiamin, lecithin, and garlic powder reacted at $140^{\circ}C$ for 30 min and were most favorable for a roasted meat flavor. Based on an omission test, cysteine was selected as the most important precursor for producing meat flavor compared to methionine, thiamine, and lecithin. Natural precursors including mushroom powder and fat medium were applied to compensate for the synthetic precursors. The optimum formula for meat flavor was 5% ribose, 7.7% cysteine, 6.9% garlic juice powder, 2.1% Lentinusedodes powder digested with protease, and 1% lard. The sulfuric pungent, oily, and salty attributes of the formula decreased and a mild roasted meat flavor was expressed.

Comparison of antioxidant capacity of protein hydrolysates from 4 different edible insects (식용곤충별 단백가수분해물의 항산화 활성 비교)

  • Jang, Hyun-Young;Park, Chae-Eun;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.480-485
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    • 2019
  • The present study was conducted to compare antioxidant capacities of protein hydrolysates from four different edible insects (Protaetia brevitarsis larvae, Allomyrina dichotoma larvae, Gryllus bimaculatus imago, and Tenebrio molitor larvae) which have recently been registered as food varieties in Korea. Protein hydrolysates were prepared from each insect using enzymatic hydrolysis using alcalase, and were then separated into a fraction containing ${\leq}3kDa$. According to $RC_{50}$ values and trolox equivalent antioxidant capacity results obtained from five different antioxidant analyses, the Gryllus bimaculatus (GB) hydrolysate showed relatively high levels of antioxidant capacity and, in particular, the GB hydrolysate showed considerably strong antioxidant activities in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and in ferric reducing antioxidant power (FRAP) assays. The GB hydrolysate also showed the strongest inhibitory effect on peroxidation of linoleic acid, and its rate of inhibition at $100{\mu}g/mL$ on day 3 of treatment was 60.26%. These results suggest that protein hydrolysates from edible insects including GB represent potential sources of natural antioxidants.

Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat (방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현)

  • Kwag, Hyon-Joo;Lee, Kyoung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.66-74
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    • 2003
  • Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.