• Title/Summary/Keyword: 기질단백분해효소

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The effect of indomethacin on the matrix metalloproteinases in canine permanent tooth eruption (인도메타신의 투여가 치아 맹출 시 기질금속단백분해 효소의 분포에 미치는 영향)

  • Kang, Yoon-Goo;Nam, Jong-Hyun;Lee, Ki-Soo
    • The korean journal of orthodontics
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    • v.36 no.2 s.115
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    • pp.91-102
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    • 2006
  • Tooth eruption requires remodeling of surrounding tissues. This study was aimed to investigate the effect of indomethacin on the dental follicle and paradental tissues during tooth eruption by observing the distribution and expression of MMP by the immunohistochemical method. Ten mongrel dogs of ten to twelve weeks old were divided into 5 groups; four experimental groups administered indomethacin 2 mg/Kg/day and 8 mg/Kg/day orally 2 times a day for 14 days and 7 days respectively, and the control group was administered a placebo. Permanent teeth before eruption and their surrounding tissues were selected and excised. H&E staining and immunohistochemical stainings of MMP-3 and -9 were performed and examined under the light microscope. Osteoclasts, osteoblasts, periodontal ligament cells, ameloblasts and odontoblasts of the control group all expressed MMP-3 and -9. In the experimental group, osteoclasts, osteoblasts and periodontal ligament cells showed reduced expression of MMP-3 and -9. Magnitude of MMP reduction In the experimental group showed a time and dose of indomethacin administration dependent manner. These results show that indomethacin inhibited MMP-3 and -9 expression in the dental follicle and surrounding tissues and suggest that when indomethacin is administered for long periods, tooth eruption could be delayed.

Enzymatic Modification of Soy Proteins: Effects of Functional Properties of Soy Isolate upon Proteolytic Hydrolysis (대두단백질(大豆蛋白質)의 효소적(酵素的) 변형(變形) : 분리대두단백질(分離大豆蛋白質)의 기능성(機能性)에 미치는 단백질가수분해(蛋白質加水分解)의 영향(影響))

  • Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.211-217
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    • 1984
  • To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

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Characterization of the Strong Proteolytic Bacteria Isolated from Low Salt Fermented Anchovy and of Protease Produced by that Strain (저식염멸치젓에서 분리한 단백질분해력이 강한 세균 및 생산된 단백분해효소의 특성)

  • CHA Yong-Jun;LEE Eung-Ho;LEE Kang-Hee;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.71-79
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    • 1988
  • For the purpose of producing low salt fermented anchovy by accelerated method with a strong proteolytic bacteria, in this study, a strong proteolytic bacterium was isolated from low salt fermented anchovy and its bacteriological characteristics and properties of protease were experimented. The results obtained were as fellows : three proteolytic bacteria, Aeromonas anaerogenes Barillus subtilis and Staphylococcus saprophyticus were isolated from low salt fermented anchovy($4\%\;of\;salt,\;4\%\;of\;KCl,\;0.5\%\;of\;lactic\;acid,\;6\%$of sorbitol and $4\%$ of alcohol extract of red pepper) after 40 days fermentation. Among these strains, which grow best at $30^{\circ}C$, pH 7.0, B. subtilis was found the best proteolytic strain and benefit for industrial use as shown $0.95\;hr^{-1}$ of specific growth rate, $89{\mu}g-Tyr/hr.ml$ of maximum activity after 12 hrs culture in TPY broth. The protease produced by by B. subtilis showed maximum activity at $35^{\circ}C$, pH 7.0, and molecular weight was estimated to be 23,000 by Sephadex G-100 filtration, and it was supposed to be a kind of metal chelator sensitive neutral protease from the results of strong sensitivity against EDTA, o-phenanthroline and metal ions such as $Cu^{2+},\;Ni^{2+},\;Fe^{2+}.Km$ value of that by method of Lineweaver-Burk was determinded to be $0.73\%$ for casein as a substrate.

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Characteristics and Action Pattern of Polygalacturonase from Penicillium sp.CB-20 (Penicillium sp. CB-20이 생성하는 Polygalacturonase의 특성 및 작용양상)

  • Cho, Young-Je;Ahn, Bong-Jeon;Lim, Seong-Il;Lee, Woo-Je;Choi, Cheong
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.580-586
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    • 1989
  • Penicillium sp. CB-20 was selected for strong polygalacturonase activity among various strains of molds found in soil. The optimum pH for the enzyme activity was 5.0 and optimum temperature was 4$0^{\circ}C$. The enzyme was relatively stable in acidic condition and unstable by heat treatment. The activation energy, Km and V$_{max}$ for the polygalacturonase were 2.499 Kcal/mol, 2.13$\times$10$^{-2}$mol/l, and 104.17 $\mu$mol/min. The activity of polygalacturonase was inhibited by Ag$^{+}$, Cu$^{++}$, Pb$^{++}$, Fe$^{+++}$, $Ca^{++}$, Na$^+$, Mn$^{++}$. The enzyme can be inactivated by the treatment ethylenediamintetra acetic acid, 2,4-dinitrophenol and $H_2O$$_2$. The results indicate the possible involvement of histidine, chelate and terminal amino group as active site. The enzyme was endo-type polygalacturonase.

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Characteristics and Action Pattern of Polygalacturonase from Rhizopus oryzae CJ-2114 (Rhizopus oryzae CJ-2114가 생성하는 Polygalacturonase의 특성 및 작용양상)

  • Chung, Yung-Gun;Cho, Young-Je;Kwon, Oh-Jin;Choi, Cheong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.2
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    • pp.195-200
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    • 1992
  • Rhizopus oryzae CJ-2114 was selected for its strong polygalacturonase activity among various strains of mold found in soil. The optimum pH for the enzyme activity was 4.0 and optimum temperature was 4$0^{\circ}C$. The activation energy for the polygalacturonase was calculated by Arrhenius equation was 2.048㎉/㏖. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the Km value of 54.05mM with the $V_{max}$ of 13.9m mole/min. The enzyme is relatively stable in acidic condition. The activity of polygalactur-onase was inhibited completely by C $u^{2+}$, P $b^{2+}$ and Z $n^{2+}$, $_Mn^{2+}$ at concentration of 1 mM. The enzyme can be inactivated by the treatment with maleic anhydride and iodine. The results indicate the possible involvement of histidine at active site. When polygalacturonase from Rhizopus oryzae CJ-2114 was reacted with poly-galacturonic acid as a substrate mono-, di-, and oligogalacturonic acid were produced at early and mono-, digalacturonic acid produced at late incubation time. time.

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Bacillus licheniformis SSA3-2M1 이 생산하는 Proteinases

  • 장영채;이경형;김성영;조윤래;김종규
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.239-245
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    • 1992
  • Buci1llr.s 11c~h~n~1rnSiSi.As 3-2MI which is responsible for the special taste of traditionalKorean soy sause produced two kinds of proteinase. The activity of the proteinasc I washigher about two fold than that of proteinase 11. The optimai, reaction pH of proteinaseI and I1 wcre found to be 7-1 1.5 and 7-9. respectively. Proteinase I1 was more stable andactive than proteinase I at pH ranges around 3 to 5. The optimal te~tlperature of proteinaseI and I1 were 502. The temperature stabilitl of proteinase I1 was Inore stable thanproteinase 1 at temperature range around 30-quot;~A. ctivities of proteinase I and I1 graduallydeclined above $30^{\circ}$C and 45C. respectively. Proteinasc 1 was more active than proteinaseI1 at salt concentration range around 25-3500. The K,,, values of casein and soy proteinfor proteinase I were 6.89 mglml and 3.98 mglml. In case of proteinase 11. they were 9.00mgiml anti 11.44 111g/ml. respectively. The activity of the crudc enzyme was increased by1 rnM Pb(CH3COO). but was decreased by 5 n1M and 10 rnM of HgS04 and ZnS04. Thetwo proteinases produced amino acids and peptides from the soybean protein. The peptideswere digested into amino acids. Both protcinases were found to be the main enzymes thatproduced amino acids which make the main taste of traditional Korean soy sauce.al Korean soy sauce.

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Anti-sclerotic Effect of Cinnamomi Ramulus Via Suppression of MMP-9 Activity and Migration of TNF-$\alpha$-induced HASMC (인간대동맥평활근의 유주능 및 기질금속단백분해효소의 억제를 통한 계지의 항동맥경화능)

  • Kim, Jai-Eun;Lee, Chang-Sup;Choi, Sung-Kyu;Choi, Dall-Yeong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.974-979
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    • 2009
  • Proliferation of vascular smooth muscle cell(VSMC) is one of the key features in onset of atherosclerosis and restenosis after vascular surgery such as stent implant. Atherosclerotic plaques are usually composed of collagen, elatsin and smooth muscle cells. Release of matrix metalloproteinases(MMPs) is considered to have correlation with development of atherosclerotic plaques. Based on the hypothesis that MMP inhibition would be helpful in the treatment of atherosclerosis, we investigated inhibition of MMP activity and migration of TNF-$\alpha$-induced human aortic smooth muscle cell(HASMC) by Cinnamomi Ramulus(CC). The result from gelatin zymography showed that CC inhibited MMP-9 activity in a dose-dependent manner. In addition, CC considerably inhibited the migration of HASMC induced by TNF-$\alpha$, while it showed little cytotoxic effect on HASMC. These results suggest that CC can be a potential anti-atherosclerotic agent through inhibition of MMP-9 activity and SMC migration.

MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide (Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구)

  • Yang, Won-Kyung;Lee, Woo-Cheol;Kim, Mi-Ri;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.372-384
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    • 2005
  • The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

Ferment Red Ginseng Suppresses the Expression of Matrix Metalloproteinases in UVA-irradiated Human Dermal Fibroblast Cells (발효홍삼의 인간진피섬유모세포에서 UVA로 유도한 염증 및 기질단백분해효소 발현 억제 효능)

  • Lee, Keun-Hyeun;Jeong, Seung-Il;Lee, Chang-Hyun;Shin, Sang Woo;Jeong, Han-Sol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.2
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    • pp.105-110
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    • 2017
  • Prolonged exposure to solar ultraviolet A (UVA) radiation has been known to cause premature skin aging (photo-aging). UVA radiation generates ROS thereby induce degenerative changes of skin such as degradation of dermal collagen, elastic fibers. Matrix metalloproteinases (MMPs), the proteolytic enzymes have been implicated as a major player in the development of UVA-induced photo-aging. Many studies have been conducted to block the harmful effects of UV radiation on the skin. Recently, we are interested in the availability of fermented red ginseng (FRG) as natural matrix metalloproteinases inhibitors (MMPIs). The efficacy difference between red ginseng and FRG has been compared. Both RG and FRG have no cytotoxic effects below the concentration of $300{\mu}g/ml$. Human dermal fibroblasts (HDFs) were pretreated with FRG or RG for 24h, followed by irradiation of UVA. Then, we measured the intracellular ROS production and the expression of MMP, $IL-1{\beta}$ at the mRNA level. We also examined the intracellular localization of $NF-{\kappa}B$ and MMP-9 on the FRG or RG treated and UVA-irradiated HDFs. FRG decreased the intracellular ROS production elicited by UVA. In addition, FRG decreased the mRNA expression of MMP-3, MMP-9, and $IL-1{\beta}$ more efficiently than RG. Furthermore, FRG suppressed the nuclear localization of $NF-{\kappa}B$, and the expression of MMP-9. Taken together, our results suggest that FRG is promising agents to prevent UVA-induced photo-aging by suppressing MMP expression and inflammation.

Activity of Matrix Metalloproteinase-2 and its Significance after Resection of Stage I Non-small Cell Lung Cancer (제1기 비소세포폐암 환자의 수술적 절제 후 Matrix Metalloprotainase-2 활성도에 따른 재발 및 예후)

  • Kim Sang Hui;Hong Young-Sook;Lee Jinseon;Son Dae-Soon;Lim Yu-Sung;Song In-Seung;Lee Hye-Sook;Kim Do Hun;Kim Jingook;Choi Yong Soo
    • Journal of Chest Surgery
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    • v.38 no.1 s.246
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    • pp.38-43
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    • 2005
  • Matrix metalloproteinase-2 (MMP-2) is a class of proteolytic enzymes that digest collagen type IV and other components of the basement membrane. It plays a key role in the local invasion and the formation of distant metastases by various malignant tumors. The aim of this study was to evaluate the activity of MMP-2 and its significance as a prognostic marker in resected stage I non-small cell lung cancer (NSCLC). Material and Method: In this study we obtained fresh-frozen samples of tumor and non-tumor tissues from 34 patients with stage I NSCLC who underwent resection without preoperative radiotherapy or chemotherapy. After the extraction of total protein from tissue samples, MMP-2 activities were assessed by gelatin-substrate-zymography. The activities were divided into the higher or lower groups. Result: The MMP-2 activities were higher in tumor tissues than in non-tumor tissues. The MMP-2 activity of non-tumor tissues in recurrent group was higher than in non-recurrent group (p<0.01). Also the patients with higher MMP-2 activity of non-tumor tissues showed poor 5 year survival (p<0.01). Conclusion: This result indicates that the higher level of MMP-2 activity in the non-tumor tissue is associated with the recurrence and survival after the resection of stage I NSCLC. Therefore, MMP-2 activity in the non-tumor tissue could be used as a potential prognostic marker for the resected stage I-NSCLC.