• Journal of Life Science
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• v.15 no.2 s.69
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• pp.231-235
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• 2005

Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-dependent enzymes responsible for proteolytic degradation of extracellular matrix components such as base membrane or interstitial stroma. MMPs play an important role in a variety of physiological and pathological tissue remodeling processes, including wound healing, embryo implantation, tumor invasion and metastasis. Since MMP-9 (gelatinase B) has unique ability to cleave type IV collagen, gene expression of MMP-9 has been focused on as a pharmacological target. Flavonoids are a class of compounds that are widely spread in plants. In the coures of screening for the suppressors of MMP-9 gene expression from natural products, Metasequoia glyptostroboides was selected. Six flavonoids, sciadopitysin, isoginkgetin, bilobetin, 2,3-dihydrohinokiflavone, luteolin and apigenin were purified as suppressors of MMP-9 gene expression from M. glyptostroboides. The suppressing activity of the isolated flavinoids on the MMP-9 gene expression was measured by gelatin zymography and Nothern blot analysis.

• Korean Journal of Food Science and Technology
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• v.18 no.4
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• pp.270-277
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• 1986

Ficin, a proteolytic enzyme in Fig latex, was extracted and purified with using ammonium sulfate and CM-cellulose column chromatography, respectively, and studied for its chemical properties. The disc gel electrophoresis showed one major and three minor bands for $(NH_4)_2SO_4$ extract and only one band showed after CM-cellulose chromatography. The optimum conditions for ficin activity was found to be pH 7.0 and $50^{\circ}C$. The amino acids composition of the purified ficin were 21.8% as acidic, 3.5% as basic and 74.7% as neutral amino acids. The amino acids analysis indicated that the ficin was composed of 174 amino acids residue having molecular weight of 19,500.

• Applied Biological Chemistry
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• v.23 no.2
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• pp.115-122
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• 1980

An attempt was made to isolate suitable kinds of fungi for the preparation of Sufu, one of the fermented soybean products, and also to investigate the chemical changes during the aging period of Sufu. 1. First, from the isolated fungi 14 strains of Mucor sp. were selected on the basis of their morphological texture and agreeable odor. Secondly, 7 strains of these fungi with relatively high proteolytic activity were selected. 2. All of the pehtzes that were fermented with the selected fungi have raised the water soluble protein content to about 10% on average. 3. The amount of hydrolyzed protein increased to the 18th. day and then did not show any visible increase during the aging period. Finally, the 3 strains of suitable fungi for preparing Sufu could be selected. It seemed that one of them was probably better than the other two which were almost same as the standard fungi, Actinomucor elegans, in protein hydrolyzation.

• Journal of the Korean Wood Science and Technology
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• v.35 no.6
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• pp.159-165
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• 2007

An extracellular xylanase from the brown-rot fungus Fomitopsis palustris grown on rice straw culture was purified to a single protein band. On SDS-PAGE, the molecular mass of purified enzyme was estimated to be about 43 kDa. The amino acid sequence of the proteolytic fragments showed high homology with fungal glycoside hydrolase family 10 xylanases. The $K_m$, $K_{cat}$ and $V_{max}$ for birch xylan were $31mg/m{\ell}$, $2.3{\times}10^4/min$ and 252.3 U/mg, respectively. The optimal activity of the purified xylanase from F palustris was observed at pH 4.0~5.0 and $70^{\circ}C$.

• The Korean Journal of Food And Nutrition
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• v.23 no.3
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• pp.399-404
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• 2010

This study was conducted to determine the influence of culture conditions such as temperature, time, water content, koji-thickness, and agitation on the production of protease by Rhizopus sp. ZB9, isolated from Korean Nuruk, during the preparation of rice koji, which is used in brewing the Korean rice wines, Takju and Yakju. Rice koji was made under different culture conditions, and the proteolytic activity of each koji was tested. The temperature range suitable for the production of protease was $28~32^{\circ}C$. Based on the protease and color, 60 hours of cultivation at $28^{\circ}C$ was shown to produce optimum results. The production of protease increased in proportion to the increase in water content of steamed rice from 25% to 35%. An increase in koji-thickness induced no adverse effects on the production of protease, and agitation during cultivation showed beneficial effects.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1252-1258
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• 2018

Objective: This study was carried out to determine the effects of cumin essential oil on the silage fermentation, aerobic stability and in vitro digestibility of alfalfa silages. Methods: Alfalfa was harvested at early bloom (5th cutting) stage in October and wilted for about 3 hours. The research was carried out at three groups which were the control group where no additive control was done (CON), cumin essential oil (CMN3) with 300 mg/kg and CMN5 with 500 mg/kg cumin essential oil addition. Alfalfa was ensiled in plastic bags. The packages were stored at $8^{\circ}C{\pm}2^{\circ}C$ under laboratory conditions. All groups were sampled for physical, chemical and microbiological analysis 120th day after ensiling. At the end of the ensiling period, all silages were subjected to an aerobic stability test for 7 days. In addition, enzimatic solubility of organic matter (ESOM), metabolizable energy (ME), and relative feed value (RFV) of these silages were determined. Results: pH level decreased in the cumin groups compared to CON (p<0.05), thus inhibiting proteolytic enzymes from breaking down proteins into ammonia. In addition, it increased ESOM amount, and concordantly provided an increase of ME contents. Similarly, dry matter intake and RFV ratio increased. After opening the silage, it kept its aerobic stability for three days. Conclusion: Cumin essential oil improved fermentation, and affected chemical and microbiological characteristics of silages. Especially the addition of 300 mg/kg cumin provided cell wall fractionation through stimulating the activities of enzymes responsible. It also increased the number and activity of lactic acid bacteria (LAB) through providing a development of LAB.

• Korean Journal of Agricultural Science
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• v.40 no.3
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• pp.221-226
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• 2013

Degrees of hydrolysis by alkaline protease produced from Serratia marcescens S3-R1 is 3.95-6.30% of whey proteins during 5, 15, 30, 60, 90, 120,180, 240 min incubation at $40^{\circ}C$. Proteolytic pattern of the whey proteins showed that various low molecular weight peptides were generated during the incubation periods. The biological function of in Raw 264.7 cells treated with whey protein hydrolytic peptides, anti-inflammatory effect showed exhibit in the expression of pro-inflammatory cytokines such as TNF-${\alpha}$, IL-6, COX-2 and iNOS by PCR analysis. COX-2 and iNOS gene expression inhibited in Raw 264.7 cells on whey protein hydrolysates below 3,000 dalton. The protease from Serratia marcescens S3-R1 showed a potential in production of low molecular weight whey protein hydrolysates which could be used for industrial application.

• Molecules and Cells
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• v.26 no.1
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• pp.41-47
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• 2008

Mitogen-activated protein kinase (MAPK) signaling is a crucial component of eukaryotic cells; it plays an important role in responses to extracelluar stimuli and in the regulation of various cellular activities. The signaling cascade is evolutionarily conserved in the eukaryotic kingdom from yeast to human. In response to a variety of extracellular signals, MAPK activity is known to be regulated via phosphorylation of a conserved $T{\times}Y$ motif at the activation loop in which both threonine and tyrosine residues are phosphorylated by the upstream kinase. However, the mechanism by which both residues are phosphorylated continues to remain elusive. In the budding yeast, Saccharomyces cerevisiae, Fus3 MAPK is involved in the mating signaling pathway. In order to elucidate the functional mechanism of MAPK activation, we quantitatively profiled phosphorylation of the $T{\times}Y$ motif in Fus3 using mass spectrometry (MS). We used synthetic heavy stable isotope-labeled phosphopeptides and nonphosphopeptides corresponding to the proteolytic $T{\times}Y$ motif of Fus3 and accompanying data-dependent tandem MS to quantitatively monitor dynamic changes in the phosphorylation events of MAPK. Phosphospecific immunoblotting and the MS data suggested that the tyrosine residue is dynamically phosphorylated upon stimulation and that this leads to dual phosphorylation. In contrast, the magnitude of threonine phosphorylation did not change significantly. However, the absence of a threonine residue leads to hyperphosphorylation of the tyrosine residue in the unstimulated condition, suggesting that the threonine residue contributes to the control of signaling noise.

• Molecules and Cells
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• v.24 no.2
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• pp.224-231
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• 2007

$H_2O_2$, as an example of oxidative stress, induces cardiac myocyte apoptosis. Bcl-2 family proteins are key regulators of the apoptotic response while their functions can be regulated by post-translational modifications including phosphorylation, dimerization or proteolytic cleavage. In this study, we examined the role of various protein kinases in regulating total BAD protein levels in adult rat cardiac myocytes undergoing apoptosis. Stimulation with 0.1 mM $H_2O_2$, which induces apoptosis, resulted in a marked down-regulation of BAD protein, which is attributed to cleavage by caspases since it can be restored in the presence of a general caspase inhibitor. Inhibition of PKC, p38-MAPK, ERK1/2 and PI-3-K did not influence the reduced BAD protein levels observed after stimulation with $H_2O_2$. On the contrary, inhibition of PKA or specifically $PKC{\delta}$ resulted in up-regulation of BAD. Decreased caspase 3 activity was observed in $H_2O_2$ treated cells after inhibition of PKA or $PKC{\delta}$ whereas inhibition of PKA also resulted in improved cell survival. Furthermore, addition of okadaic acid to inhibit selected phosphatases resulted in enhanced BAD cleavage. These data suggest that, during oxidative stress-induced cardiac myocyte apoptosis, there is a caspase-dependent down-regulation of BAD protein, which seems to be regulated by coordinated action of PKA, $PKC{\delta}$ and phosphatases.

• International Journal of Oral Biology
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• v.35 no.4
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• pp.153-158
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• 2010

Angelica decursiva has been used in Korean traditional medicine as an antitussive, an analgesic, an antipyretic and a cough remedy. However, the anti-cancer properties of Angelica decursiva have not yet been well defined. In our current study the cytotoxic activity of ethanol extracts of Angelica decursiva root (EEAD) and the mechanism of cell death exhibited by EEAD were examined in FaDu human head and neck squamous cell carcinoma cells. The cytotoxic effects of EEAD upon the growth of FaDu cells were examined with an MTT assay. In addition, the mechanism of cell death induced by EEAD was evaluated by DNA fragmentation analysis, immunoblotting and caspase activation measurements. EEAD induced apoptotic cell death in FaDu cells in a concentration- and time-dependent manner, as determined by MTT assay and DNA fragmentation analysis. Furthermore, the proteolytic processing of caspase-3, -7 and -9 was increased by EEAD treatment of FaDu cells. In addition, the activation of caspase-3 and -7 was detected in living FaDu cells by fluorescence microscopy. These results suggest that EEAD can induce apoptosis and suppress cell growth in cancer cells and may have utility as a future anticancer therapy.