• Title/Summary/Keyword: protease inhibition

Search Result 218, Processing Time 0.02 seconds

Nutritional Value and Bioactive Properties of Enzymatic Hydrolysates prepared from the Livers of Oncorhynchus keta and Oncorhynchus gorbuscha (Pacific Salmon)

  • Yoon, Ho Dong;Karaulova, Ekaterina P.;Shulgina, Lilia V.;Yakush, Evgeni V.;Mok, Jong Soo;Lee, Su Seon;Xie, Chengliang;Kim, Jeong Gyun
    • Fisheries and Aquatic Sciences
    • /
    • v.18 no.1
    • /
    • pp.13-20
    • /
    • 2015
  • Calculated chemical scores (computed in relation to the FAO/WHO reference protein) for salmon liver protein hydrolysates indicated that all amino acids (other than methionine and threonine) were present in adequate or excess quantities; thus, the raw liver material is a good source of essential amino acids. The hydrophobic amino acids contents in hydrolysates prepared from Oncorhynchus keta and O. gorbuscha were 38.4 and 39.1%, respectively. The proportion of released peptides exceeding 500 kDa was reduced when hydrolysates were treated with the commercial enzyme Alcalase, although proportions in the following MW ranges were elevated: 100-500 kDa and <50 kDa. The optimal conditions for enzymatic hydrolysis were as follows: pH 7.0, $50^{\circ}C$, and a reaction time of 1 h. Of the different proteases tested, Alcalase was the most efficient for production of salmon liver hydrolysate with the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity. The hydrolysates prepared from salmon liver had a balanced amino acid composition. The liver protein hydrolysates contained low molecular weight peptides, some of which may be bio-active; this bio-active potential should be investigated. Inhibition of the DPPH radical increased with increased degree of hydrolysis (DH), regardless of protease type. DPPH radical scavenging abilities, antithrombotic effects and ${\alpha}$-glucosidase enzyme inhibition effects of O. keta liver hydrolysate increased in a dose-dependent manner. Thus, salmon liver hydrolysate may be useful in functional food applications and as a source of novel products.

Inhibitory effect of Ulmus davidiana Planch extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts

  • Park, Jun-Sung;Kim, Kyung-Ho;Jo, Hyun-Seog;Kim, Kap-Sung;Hwang, Min-Seob
    • Journal of Acupuncture Research
    • /
    • v.22 no.2
    • /
    • pp.55-70
    • /
    • 2005
  • Objective: Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammnatory in the traditional Korean medicine. UD has been reported as a good enhancer for bone healing. Methods : In this experiment, we investigate the Inhibitory effects of UD on bone resorption using the bone cells culture. Different concentrations of crude extract of UD were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MIT assay. It was demonstrated that UD has potential effects on bone cells culture without any cytotoxicity. The most effective concentration of UD on bone cells were $100\;{\mu}g/ml$. Cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. Results : When mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-Kinase inhibitor, wortmannin (WT), WT prevented the osteoclast-mediated intracellular processing of Cat K. Similarly, treatment of osteoclasts-containing long bone cells with UD extracts prevented the intracellular maturation of Cat K, suggesting that UD may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose-6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and UD in the absence or presence of M6P. Inhibition of Cat K processing by WT or UD was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and UD. Conclusion : UD dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing.

  • PDF

A Caspase Inducing Inhibitor Isolated from Caesalpinia sappan (소목(Caesalpinia sappan)으로부터 분리한 caspase 유도 저해 물질)

  • Son, Eun-Jung;Kim, Jin-Hee;Kim, Hyun-A;Baek, Seung-Hwa;Kho, Yung-Hee;Kim, Mee-Ree;Lee, Choong-Hwan
    • Korean Journal of Food Science and Technology
    • /
    • v.35 no.4
    • /
    • pp.680-683
    • /
    • 2003
  • Through the screening of caspase-3 inducing inhibitors in U937 human monocytic leukemia cell from natural sources, Caesalpiniae sappan, which showed a high level of inhibition, was selected. The inhibition compound was purified from methanol extract by silica gel column chromatography and HPLC. The inhibitor was identified as brazilin by spectroscopic methods of ESI-MS, $^1H-NMR$, and $^{13}C-NMR$. Brazilin showed inhibitory activity of caspase-3 induction, a major protease of apoptosis cascade, with $IC_{50}$ value of $4.5\;{\mu}g/mL$ after 7 hr of treatment in U937 cells.

Functional Cosmetic Effect of Porcine Placeta (Porcine Placenta의 기능성 화장품소재 특성)

  • Kim, Bo Young;Kim, Tagon;Kang, Whan Yul;Baek, Hyun;Cheon, Hae Young;Kim, Donguk
    • Korean Chemical Engineering Research
    • /
    • v.48 no.3
    • /
    • pp.327-331
    • /
    • 2010
  • Porcine placenta was treated with alkali, acid and enzyme treatment to obtain extracts. Heavy metal contents such as Pb, As, and Hg were low enough to satisfy cosmetic agent standard. As a result of safety test(MTT assay), porcine placenta extracts showed over 80% of cell viability at $50{\mu}g/ml$, and cell toxicity was relatively lower. From antioxidation test using DPPH free radical scavenging assay, antioxidation effect was highest as 63% at $50{\mu}g/ml$ when porcine placenta was treated with alkali in pH 9. From whitening effect test using tyrosinase inhibition assay, tyrosinase inhibition effect was 30% at $50{\mu}g/ml$ concentration in alkali treated procine placenta, however, the efficiency was lower compared with arbutin or vitamin C. In anti-wrinkle effect test from elastase inhibition assay, elastase inhibition effects were 20~30% at $50{\mu}g/ml$ for 5 kinds of porcine placenta treatments, which was superior to standard, and especially, protease treated extracts showed best results. Skin formulation including 1% porcine placenta was made and the formulation was very stable for temperature and storage period. From this research, porcine placenta extract showed high potential for anti-wrinkle functional cosmetic agent.

The Protective Effects of Sasim-tang on the Brain Cell Damage in Photothrombotic Ischemia Mouse Model (광화학적 허혈성 뇌졸중 모델에서 사심탕(瀉心湯)의 뇌세포 손상 보호효과)

  • Kang, Baek-Gyu;Yun, Jong-Min;Moon, Byung-Soon
    • The Journal of Internal Korean Medicine
    • /
    • v.33 no.4
    • /
    • pp.572-586
    • /
    • 2012
  • Objectives : This study was designed to investigate the effects of Sasim-tang (SST) on proinflammatory cytokine production in a photothrombotic ischemia mouse model. Methods : Photothrombotic ischemia was induced in stereotactically held male Balb/c mice using rose bengal (10 mg/kg) and cold light. The target of photothrombotic ischemic lesion was 1 mm anterior to bregma and 3 mm lateral to midline with 2 mm in diameter, which are decreased by oral administration of SST. Results : SST protected ischemic death of brain cells through inhibition of pro-inflammatory cytokines production and catalytic activation of caspase-3 protease in photothrombotic ischemia mouse model. Conclusions : The results of this study suggest that SST can have protective effects on brain cell damage in a photothrombotic ischemia mouse model.

Protective Effects of Jihwangeumja on Oxidative Stress-induced Injury of Human Umbilical Vein Endothelial Cells (혈관내피세포의 산화적 손상에 대한 지황음자의 방어기전 연구)

  • 정용준;장재호;이대용;이민구;전인철;정대영;이인;신선호;문병순
    • The Journal of Korean Medicine
    • /
    • v.25 no.2
    • /
    • pp.173-183
    • /
    • 2004
  • Objectives : Oxidative stress can induce negative responses such as growth inhibition or cell death by necrosis or apoptosis due to the intensity of the oxidative stress, as well as positive responses such as cellular proliferation or activation. We examined the effect of Jihwangeumja on this process. Methods and Results : We analyzed the influence of oxidative stress and agents that modify its effect in human umbilical vein endothelial cell (HUVEC). Oxidative stress was induced by $B_2O_2$. With induced oxidative stress the results obtained indicate that it has a harmful effect over cell function and viability, and that this effect is dose and time dependent. When oxidative stress increased, Jihwangeumja reduced cell damage and had protective functions. $B_2O_2$, induced the apoptosis of HUVEC through the activation of intrinsic caspases pathway as well as mitochondrial dysfunction. A significant increase in cell survival was observed in culture cells with oxidative stress when they were treated with Jihwangeumja. Conclusions : These results suggest that Jihwangeumja may be potentially useful to treat HUVEC against oxidative damages mediated by modulation of caspase protease and mitochondrial dysfunction.

  • PDF

Identification and Partial Characterization of Lacticin SA72, a Bacteriocin Produced by Lactococcus lactis SA72 Isolated from Jeot-gal

  • Koo, Kyoung-Mo;Lee, Na-Kyoung;Hwang, Young-Il;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • v.10 no.4
    • /
    • pp.488-495
    • /
    • 2000
  • Strain SA72 was isolated from Jeot-gal and identified as producer of a bacteriocin, which showed some bactericidal activity against Lactobacillus delbrueckii ATCC 4797. Strain SA72 was tentatively identified as Lactococcus lactis according to the AOI test. Lactococcus lactis SA72 showed a broad spectrum of microorganisms, tested by the modified deferred method. The activity of lacticion SA72, named tentatively as a bacteriocin produced by Lactococcus lactis SA72, was detected during the mid-lon growth phase, reached a maximum during the early stationary phase, and then declined after the late stationary phase. Lacticin SA72 also showed a relatively broad spectrum of activity against non-pathogenic and pathogenic microorganisms when assessed by the spot-on-lawn method. Its anitimicrobial activity on sensitive indicator cells disappeared completely by protease XIV treatment. The inhibitory activity of lacticin SA72 remained after treatment for 15 min at $121^{\circ}C$, 문 was stable in a pH range of 2.0 to 9.0 and all organic solvents examined. It demonstrated a typical bactericidal mode of inhibition against Lactobacillus delbrueckii ATCC 4797. The apparent molecular mass of lacticin SA72 was in the region of 3-3.5 kDa, determined by SDS-PAGE.

  • PDF

재래식 메주에서 분리한 효모들의 각종 효소활성과 가능성

  • Lee, Jong-Soo;Yi, Sung-Hun;Kwon, Su-Jin;Ahn, Cheol;Yoo, Jin-Young
    • Microbiology and Biotechnology Letters
    • /
    • v.25 no.5
    • /
    • pp.448-453
    • /
    • 1997
  • Enzyme activities, production of killer toxin and some functionality of forty seven yeasts isolated from traditional Meju were investigated in culture broth and cell free extracts. Activities of $\alpha$-galactosidase, invertase and inulinase were detected in cell free extracts of 38 strains, 43 strains and 45 strains, respectively and acidic and neutral protease activities also were detected in culture broth of all the strains, $\beta$-Galactosidase activity was detected in cell free extracts of OE-20 and S-14 strains. Killer toxins were produced by OE-12, S-8 (Candida spp.), OE-19 (Zygosaccharomyces spp.) and S-3 (Saccharomyces spp.). Culture broth of OE-23 and S-9 showed 61.3% and 59.2% of antioxidant activity to $\alpha$, $\alpha$-diphenyl-$\beta$-picrylhydrazyl(DPPH), but nitrite-scavenging ability as well as inhibition of tyrosinase and polyphenol oxidase were not appeared in all the strains.

  • PDF

Hepatitis E Virus Papain-Like Cysteine Protease Inhibits Type I Interferon Induction by Down-Regulating Melanoma Differentiation-Associated Gene 5

  • Kim, Eunha;Myoung, Jinjong
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.11
    • /
    • pp.1908-1915
    • /
    • 2018
  • Upon viral infection, the host cell recognizes the invasion through a number of pattern recognition receptors. Melanoma differentiation associated gene 5 (MDA5) and retinoic acid-inducible gene-I (RIG-I) recognize RNA molecules derived from invading viruses, activating down-stream signaling cascades, culminating in the induction of the type I interferon. On the other hand, viruses have evolved to evade type I interferon-mediated inhibition. Hepatitis E virus has been shown to encode a few antagonists of type I interferon and it is not surprising that viruses encode multiple mechanisms of viral evasion. In the present study, we demonstrated that HEV PCP strongly down-regulates MDA5-mediated activation of interferon ${\beta}$ induction in a dose-dependent manner. Interestingly, MDA5 protein expression was almost completely abolished. In addition, polyinosinic polycytidylic acid (poly(I:C))- and Sendai virus-mediated activation of type I interferon responses were similarly abrogated in the presence of HEV PCP. Furthermore, HEV PCP down-regulates several molecules that play critical roles in the induction of type I IFN expression. Taken together, these data collectively suggest that HEV-encoded PCP is a strong antagonist of type I interferon.

Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage

  • Lee, Kang Pa;Kim, Jai-Eun;Park, Won-Hwan
    • Nutrition Research and Practice
    • /
    • v.9 no.6
    • /
    • pp.586-591
    • /
    • 2015
  • BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.