• Title/Summary/Keyword: Extracellular products

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Evaluation of Effective MMP Inhibitors from Eight Different Brown Algae in Human Fibrosarcoma HT1080 Cells

  • Bae, Min Joo;Karadeniz, Fatih;Ahn, Byul-Nim;Kong, Chang-Suk
    • Preventive Nutrition and Food Science
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    • v.20 no.3
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    • pp.153-161
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    • 2015
  • Matrix metalloproteinases (MMPs) are crucial extracellular matrices degrading enzymes that have important roles in metastasis of cancer progression as well as other significant conditions such as oxidative stress and hepatic fibrosis. Marine plants are on the rise for their potential to provide natural products that exhibit remarkable health benefits. In this context, brown algae species have been of much interest in the pharmaceutical field with reported instances of isolation of bioactive compounds against tumor growth and MMP activity. In this study, eight different brown algae species were harvested, and their extracts were compared in regard to their anti-MMP effects. According to gelatin zymography results, Ecklonia cava, Ecklonia bicyclis, and Ishige okamurae showed higher inhibitory effects than the other samples on MMP-2 and -9 activity at the concentrations of 10, 50, and $100{\mu}g/mL$. However, only I. okamurae was able to regulate the MMP activity through the expression of MMP and tissue inhibitor of MMP observed by mRNA levels. Overall, brown algae species showed to be good sources for anti-MMP agents, while I. okamurae needs to be further studied for its potential to yield pharmaceutical molecules that can regulate MMP-activity through cellular pathways as well as enzymatic inhibition.

Characteristics of Mannitol Production by Leuconostoc sp. KY-002 (Leuconostoc sp. KY-002의 Mannitol 생산 특성)

  • 류병호;김동현윤종원
    • KSBB Journal
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    • v.11 no.6
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    • pp.636-641
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    • 1996
  • The production of extracellular mannitol by a new mannitol-producing bacterium, Leuconostoc sp. KY-002 was studied in shake flask cultures. The new isolate has a capability of utilizing fructose and sucrose for mannitol formation. Maximum mannitol production was obtained with fructose as the sole carbon source. Under the optimal culture conditions, within 70 hours of incubation, a final concentration of 26 g/L of mannitol from 50 g/L fructose was obtained with an indicated yield of 52% based on fructose consumed. However, higher concentrations of fructose ranging from 100 to 250 g/L could not effectively be transformed to mannitol due to a lack of osmotolerance. The strain produced no other polyols such as glycerol and sorbitol as by-products. Yeast extract was the best nitrogen source and high levels of inorganic phosphate up to 10 g/L promoted mannitol formation. Any mineral ions and salts did not play important role in both cell growth and mannitol production. Nicotinic acid enhanced mannitol production by 16%. The optimum culture temperature and initial pH were $35^{\circ}C$ and 6, respectively.

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STUDIES OF ACNE TREATMENT USING ORIENTAL HERBS (New Approach to select anti-acne agents)

  • Chunja Nam;Y.G. Han;Kim, S.J.;Kim, J.H.;Oh, J.Y.
    • Proceedings of the SCSK Conference
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    • 1999.10a
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    • pp.111-121
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    • 1999
  • Acne vulgaris, the most common skin disease. can be formed as only a few comedons or severe inflammatory lesions. The pathogenesis of acne involves various factors; excessive androgen, excessive sebum production, abnormal alteration of follicular epithelium, proliferation of Propionibacterium acnes, and inflammation. We investigated acne therapy using oriental herbs described in the Korean traditional medical book (Dong-ui-bo-gam). Oriental herbs (Angleica daurica. Arctium lappa. Coptidis rhizoma, and glycyrrhiza glabra) were chosen based on their respective property of sebum control, anti-inflammatory activity, and anti-bacterial activity. We examined the effect of acne treatment, in terms of chemotactic inhibition, lipogenesis inhibition, and anti-bacterial activity for P. acnes. 1. Neutrophil chemotaxis assay ; P acnes secrete chemotactic factors and other pro-inflammatory extracellular products. Neutrophil chemotactic activity of P. acnes was measured by 48-well chemotaxis method. Angelica daurica clearly suppressed chemotactic activity of P. acens. 2. Using sebaceous gland of hamster ear lipogenesis assay; Sebaceous lipogenesis was measured using ear biopsies by incubation of $C^{14}$ -acetate in culture media. The $C^{14}$ -labeled lipids were extracted and determined by liquid scintilation counting, Coptidis rhizoma markedly inhibited sebum production, 3. Anti-bacterial assay for P. acnes (MIC test) Glycyrrhiza glabra showed anti-bacterial activity. P. acnes did not develop resistance against Glycyrrhiza glabra. Retinoids are effectively to inhibit sebum production and regulate follicular keratinization process, with little anti-inflammatory activity. Angelica daurica suppressed neutrophil chemotaxis. Coptidis rhizoma inhibited sebum production, and Glycyrrhiza glabra showed anti-bacterial activity against P. acnes. A combined formulation of Angelica daurica. Coptidisr hizoma and Glycyrrhiza glabra is expected to provide effective acne treatment.ent.ive acne treatment.

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The penetration enhancement and the lipolystic effects of TAT-GKH, in both In vitro, Ex vivo, and In vivo.

  • Lim, J.M.;Chang, M.Y.;Park, S.G.;Kang, N.G.;Song, Y.S.;Lee, Y.H.;Yoo, Y.C.;Cho, W.G.;Han, S.G.;Kang, S.H.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.87-107
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    • 2003
  • It was demonstrated that Transactivating transcriptional activator(TAT) protein from HIV-1 shown to enter cells when added to the surrounding media. TAT peptide chemically attached to various proteins was able to deliver these proteins to various cell and even in tissues in mice with high levels in heart and spleen. In this study, the tripeptide GKH(Glycine-Lysine-Histidine) derived from Parathyroid hormone (PTH), which was known as lipolytic peptide, is attached to 9-poly Lysine(TAT) to be used as a cosmetic ingredient for slimming products. When Glycerol release, expressed as extracellular glycerol concentration, is lipolysis index, TAT-GKH at $10^{-5}$mo1/L induces approximately 41.5% maximal lipolytic effects in epididymal adipocytes isolated from rats, compared with basal lipolysis. Epididymal adipose tissues of male rats is assessed ex vivo by microdialysis. Probes are perfused with Ringer solution in which increasing concentrations of TAT-GKH. The perfusion of TAT-GKH induces lipolytic effect. Penetration study showed that TAT-GKH efficiently elevates 36 times higher penetration into the excised hairless mice skin than GKH. in vivo study showed that TAT-GKH had a better effect upon the relative volume of eye bag after 28 days of application on twenty(+2) healthy female volunteers. It was identified that TAT-GKH increases penetration enhancement and lipolytic effects in both in vitro, ex vivo and in vivo.

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The effect of plant extracts on the activity and the expression of MMPs (matrix metalloprotease) induced by UVA

  • Lee, Dong-hwan;Lee, Bum-chun;Yoon, Eun-jeong;Lee, Kyung-eun;Park, Sung-min;Pyo, Hyeong-bae;Choe, Tae-boo
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.32-43
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    • 2003
  • UV irradiation on a skin brings about the qualitative and quantitative alterations of the extracellular matrix. Repeated-UV irradiation suppressed the synthesis of collagen and activated the expression of the matrix metalloprotease (MMP). In this paper, on the purpose of development of novel anti-aging agents from natural sources, effects of several natural products on in vitro MMP-1 activity and UVA induced MMP-1 synthesis in human dermal fibroblast (HDF) culture were studied. We measured MMP-1 activities by fluorescence assay using gelatin as substrates. As a result, the extract of Dicentra spectabilis, and flower buds of Tussilago farfara showed strong inhibitory effect. Among them, the extract of flower buds of Tussilago fartara and Dicentra spectabilis inhibited MMP-1 activity by 92% and 87% at 0.05% (w/v). And UVA induced MMP-1 expression were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in HDF culture. The extract of flower buds of Tussilago farfara and Dicentra spectabilis suppressed the UVA induced expression of MMP-1 by similar level of Vitamin C 200$\mu$M at 0.1% (w/v). These results suggest that the extract of Dicentra spectabilis, and flower buds of Tussilago farfara effectively prevent skin from the UV-induced photoaging. So the extracts are thought to have potential as effective raw materials for anti-aging cosmetics.

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Isolation and characterization of anaerobic microbes from marine environments in Korea (한반도 주변 해역으로부터 혐기성 미생물의 분리 및 분리 미생물의 특성 분석)

  • Kim, Wonduck;Lee, Jung-Hyun;Kwon, Kae Kyoung
    • Korean Journal of Microbiology
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    • v.52 no.2
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    • pp.183-191
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    • 2016
  • Marine bacteria have represented unique physiologies and products which are not discovered from terrestrial organisms. There has been great interest to utilize and develop marine bacteria in many industrial sectors. Recently, we isolated and characterized anaerobic bacteria from various marine environments in Korea to search organic acids fermenting strains. From our enrichment performed under anaerobic condition, 65 strains were isolated and identified by the 16S rRNA gene sequence analysis. Among them, eleven strains were selected for phylogenetical and biochemical analysis. All tested strains were affiliated with Class Clostridia except one with Class Bacteroidia. Most of strains produce acetate (6 strains) with butyrate (2 strains) and/or formate (4 strains). Strain MCWD5 transformed 40% of glucose to extracellular polymeric substances. These results indicate that many novel anaerobic microorganisms which have great potential in commercial application are distributed in the marine environments of Korean Peninsula.

Isolation and Identification of Xylanase Secreting Yeast (Xylanase를 분비하는 효모 균주의 분리 및 성질)

  • 배명애;서정훈
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.499-504
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    • 1988
  • Among the new yeast strains which were isolated from soils by incubating in the xylan containing minimal medium at 3$0^{\circ}C$, one strain(XB-33) was finally selected by the results of extracellular xylanase production test. The characteristics of XB-33 was almost consistent with those of the Cryptococcus ater. The formation of xylanase activity was induced by xylan and repressed by xylose or glucose. The xylanase was partially purified from the culture supernatant with DEAE-Sephadex A5O chromatography. The enzyme had a pH optimum for activity at 5.0 and its stability range was pH 5-7. The temperature optimum was at 5$0^{\circ}C$, but the enzyme activity was greatly lost by heating at 7$0^{\circ}C$ for 60 minutes. The hydrolysis products from xylan by crude enzyme detected by TLC, were xylose and n series of higher oligosaccharides. The Km value of xylanase was 20 (mg/ml).

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Purification and Characterization of Antifungal Chitinase from Pseudomonas sp. YHS-A2

  • Lee, Han-Seung;Lee, Hyun-Jung;Choi, Sung-Won;Her, Song;Oh, Doo-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.107-113
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    • 1997
  • A strain producing a high amount of chitinase was isolated from soil, identified as Pseudomonas sp., and tentatively named Pseudomonas sp. YHS-A2. An extracellular chitinase of Pseudomonas sp. YHS-A2 was purified according to the procedure of ammonium sulfate saturation, affinity adsorption, Sephadex G-100 gel filtration and Phenyl-sepharose CL-4B hydrophobic interaction column chromatography. The molecular weight of the purified enzyme was estimated to be 55 kDa on SDS-PAGE was confirmed by active staining. Optimal pH and temperature of the enzyme are pH 7.0 and $50^{\circ}C$, respectively, and the enzyme is stable between pH 5.0 and 8.0 and below $50^{\circ}C$. The main products of colloidal chitin by the chitinase were N-acetyl-D-glucosamine and N,N'-diacetylchitobiose both of which were detected by HPLC analysis. The enzyme is supposed to be a random-type endochitinase which can degrade any position of ${\beta}$-l,4-linkages of chitin and chitooligosaccharides. The chitinase inhibited the growth of some phytopathogenic fungi, Fusarium oxysporum, Botrytis cineria, and Mucor rouxii and these antifungal effects were thought to be due to the characteristics of endochitinase.

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Production and Characterization of Mannanase from a Bacillus sp. YB-1401 Isolated from Fermented Soybean Paste (된장 분리균 Bacillus sp. YB-1401의 Mannanase 생산성과 효소특성)

  • Joen, Ho Jin;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.99-105
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    • 2014
  • A Bacillus strain capable of hydrolyzing locust bean gum was isolated as a producer of extracellular mannanase by way of an enrichment culture in an acidic medium from homemade soybean pastes. The isolate YB-1401 showed a biochemical identity of 61.1% with Brevibacillus laterosporus, while the nucleotide sequence of its 16S rDNA had the highest similarity with that of Bacillus amyloliquefaciens. The mannanase productivity of the Bacillus sp. YB-1401 was drastically increased by mannans. Particularly, maximum mannanase productivity was reached at approximately 265 U/ml in LB medium supplemented with konjac glucomannan (4.0%). The mannanase was the most active at $55^{\circ}C$ and pH 5.5. Mannanase activity was completely maintained after pre-incubation at pH 3.5 to 11.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannobiose and mannotriose for LBG, guar gum or mannooligosaccharides. A small amount of mannose was also detected in the hydrolyzates.

Extracts of Aster species Inhibit Invasive Phenotype and Motility of H-ras MCF10A Human Breast Epithelial Cells Possibly via Downregulation of MMP-2 and MMP-9

  • Ahn, Seong-Min;Lee, Kang-Ro;Moon, A-Ree
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.240-245
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    • 2002
  • Cancer metastasis represents the most important cause of cancer death and antitumor agents that may inhibit this process have been extensively pursued. Invasion and metastasis of malignantly transformed cells involve degradation of the extracellular matrix (ECM) components by matrix metalloproteinases (MMP), especially MMP-2 and -9. We previously showed that H-ras-induced invasive phenotype may involve MMP-2, rather than MMP-9, in MCF10A cells. In the present study, we investigated the chemopreventive effect of Aster, a widely used culinary vegetable in Korea. We screened twelve extracts from three Aster species (Aster scaber, Aster oharai and Aster glehni) for the inhibitory effect on MMP activities of H-ras MCF10A human breast epithelial cells. All of the extracts tested in this study efficiently inhibited the gelatinolytic activities of MMP-2 and MMP-9. A more prominent inhibition was observed in MMP-2 activity compared to MMP-9. Out of twelve extracts, eight extracts showed>90% inhibition of MMP-2 activity in H-ras MCF10A cells while only one extract showed>90% inhibition of MMP-9 activity. We selected three extracts (AO-3, AG-3 and AS-EA) for further studies since they exerted a marked inhibition in the ratio of MMP-2 to MMP-9. Treatment with AO-3, AG-3 and AS-EA in H-ras MCF10A cells caused a significant inhibition of invasive phenotype and migration, proving a chemopreventive potential of these extracts. Taken together, our results demonstrate that extracts of Aster effectively inhibit invasion and migration of highly malignant human breast cells, possibly via downregulation of MMP-2 and MMP-9.