• Title/Summary/Keyword: enzyme inducer

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Novel Vancomycin Resistance System in Streptomyces coelicolor

  • Hong, Hee-Jeon
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.143-147
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    • 2005
  • The non-pathogenic, non-glycopeptide-producing actinomycete Streptomyces coelicolor carries a cluster of seven genes (vanSRJKHAX) that confers inducible, high-level resistance to vancomycin. The van genes are organised into four transcription units, vanRS, vanJ, vanK and vanHAX, and these transcripts are induced by vancomycin in a vanR-dependent manner. vanHAX are orthologuous to genes found in vancomycin resistant enterococci that encode enzymes predicted to reprogramme peptidoglycan biosynthesis such that cell wall precursors terminate in D-Ala-D-Lac, rather than D-Ala-D-Ala. vanR and vanS encode a two-component signal transduction system that mediates transcriptional induction of the seven van genes. vanJ and vanK are novel genes that have no counterpart in previously characterised vancomycin-resistance clusters from pathogens. VanK is essential for vancomycin resistance in S. coelicolor and it is required for adding Gly branch to stem peptides terminating D-Ala-D-Lac. Because VanK can recognise D-Lac-containing precursors but the constitutively expressed femX enzyme, encoded elsewhere on the chromosome, cannot recognize D-Lac-containing precursors as a substrate, vancomycin-induced expression of VanHAX in a vanK mutant is lethal. Further, femX null mutants are viable in the presence of glycopeptide antibiotics but die in their absence. Bioassay using vanJp-neo fusion reporter system also showed that all identified inducers for van genes expression were glycopeptide antibiotics, but teicoplanin, a membrane-anchored glycopeptide, failed to act as an inducer.

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Studies on the Microbial Glucose Isomerase Part 1. Isolation and Characterization of Streptomyces species Producing Glucose Isomerase (미생물의 포도당 이성화효소에 관한 연구 (제1보) 포도당 이성화효소 생산균주의 분리 및 성질에 관하여)

  • Chung, Tai-Wha;Kim, Hyun U.;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.4 no.4
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    • pp.138-144
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    • 1976
  • five strains of Streptomyces spp. with high Productivity of glucose isomerase (15-30 units/ml) were obtained among 280 microbial strains isolated from 150 soil samples. These strains produced glucose isomerase with xylose as an inducer. These 5 strains were also identified to be different strains of Streptomyces spp.:streptomyces sp. K-14, K-53, K-71, K-77 and K-733. It was found that Streptomyces sp. K-14 produced the highest enzyme activity. The spore chains of these strains were rectiflexible and spore surface was smooth except Steptomyces sp. K-77 and K-733, with spiny surface.

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Purification and Comparison of Properties of the C-Terminus Truncated Agarase of Pseudomonas sp. W7

  • Yoon, Soo-Cheol;Lee, Jong-Hee;Ahn, Sun-Hee;Lee, Eun-Mi;Park, Eun-Mi;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.767-772
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    • 2003
  • Three plasmids derived from the ${\beta}-agarase$ gene (PjaA) of Pseudomonas sp. W7 were expressed in Escherichia coli AD494(DE3) pLysS with lactose as an inducer. These products corresponded to the complete (PjaA) and the two C-terminal truncated (PjaAI and PjaAII) forms of ${\beta}-agarase$. The PjaAI and the PjaAII were originated from exonuclease L treatment from PjaA by deleting 127 and 182 amino acid residues-encoded nucleic acids at 3' region, respectively. The molecular weights of the purified proteins were 71 kDa, 58 kDa, and 50 kDa on SDS-PAGE, respectively. The $K_m$ value of PjaAI was lower than that of the PjaA, and the catalytic efficiency ($k_{cat}/K_m$) of PjaAI was increased to 5 times. The enzyme of PjaAI retained more than 90% activity at $50^{\circ}C$. In contrast to the PjaAI, the remaining activity of the PjaA was only 20% at the same temperature.

Induction of Quinone Reductase and Glutathione S-Transferase in Murine Hepatoma Cells by Flavonoid Glycosides

  • Kim, Jung-Hyun;Lee, Jeong-Soon;Kim, Young-Chan;Chung, Shin-Kyo;Kwon, Chong-Suk;Kim, Young-Kyoon;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.365-371
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    • 2003
  • The potential of seven flavonoid glycosides to induce quinone reductase (QR), an anticarcinogenic marker enzyme, in murine hepatoma cells (hepalc1c7) and its mutant cells (BPRc1) was evaluated. Among test compounds, kaempferol-3-O-glucoside, luteolin-6-c-glucoside, and quercetin-3-O-glucoside (Q-3-G) induced QR in hepalc1c7 cells in a dose-dependent manner. However, in BPRc1 cells lacking arylhydrocarbon receptor nuclear translocator (ARNT), only Q-3-G caused a significant induction of quinone reductase at the concentration range of 0.5 to 8 ug/mL, suggesting that it is a monofunctional inducer. Q-3-G induced not only phase 2 enzymes, including QR and glutathione-S-transferase, but also nitroblue tetrazolium reduction activity in HL-60 cells, a biochemical marker for cell differentiation promoting agents. In conclusion, Q-3-G merits further study to evaluate its cancer chemopreventive potential.

Effects of bacterial LPS and DNA on the induction of IL-1β, IL-10 and IL-12 by mouse peritoneal macrophages in vitro

  • Samad, D. Abdel;Abdelnoor, AM
    • Advances in Traditional Medicine
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    • v.6 no.2
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    • pp.134-143
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    • 2006
  • The capacities of bacterial DNA, extracted from Salmonella typhimurium, and lipopolysaccharide (LPS), extracted from Salmonella minnesota, to activate mouse peritoneal macrophages in vitro were compared. Activation was assessed by estimating e levels of 3 cytokines, IL-10, IL-12, and $IL-1{\beta}$, at time intervals of 3, 6, 9, and 24 h after addition of LPS and/or DNA to macrophage cultures. Cytokine levels in culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and cytokine mRNA levels were estimated based on band intensity in cultured cells by reverse transcriptase-polymerase chain reaction (RT-PCR). Results obtained demonstrated the ability of DNA and LPS to elicit increased production of all 3 cytokines as compared to controls. In the amount tested, LPS appeared to be a more potent inducer of IL-12, and $IL-1{\beta}$, whereas DNA induced higher levels of IL-10. DNA and LPS, used in combination, exhibited neither an additive nor a synergistic effect. Rather, an antagonist effect appeared to occur. RT-PCR results correlated well with ELISA.

Optimization of Bovine Testicular PH-20 hyaluronidase Production in Pichia pastoris (소의 히아론산 분해효소(PH-20)의 Pichia pastoris에서의 생산 최적화)

  • Shin, Hwa Shook;Kim, Eunki
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.764-768
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    • 2008
  • Bovine testicular hyaluronidase PH-20 was cloned into pPIC9 vector and expressed in Pichia pastoris. Recombinant PH-20 was 75 kDa MW and 7460 units/L activity. Extracellular hyaluronidase activity was two times higher than that of intracellular activity. Non-buffered medium and $30^{\circ}C$ cultivation was favorable for PH-20 production. 1M sorbitol as an osmotic pressure and 0.3% methanol inducer increased cell growth and enzyme activity. 0.4 M arginine augmentation decreased the proteolytic degradation of recombinant hyaluronidase.

Mutanase Induction in Trichoderma harzianum by Cell Wall of Laetiporus sulphureus and its Application for Mutan Removal from Oral Biofilms

  • Wiater, Adrian;Szczodrak, Janusz;Pleszczynska, Malgorzata
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1335-1341
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    • 2008
  • The cell wall material from fruiting bodies of Laetporus sulphureus has been suggested as a new alternative to mutan for the mutanase induction in Trichoderma harzianum. Structural analyses revealed that the cell wall fraction from this polypore fungus contained 56.3% of (1$\rightarrow$3)-linked $\alpha$-glucans. When the strain T. harzianum F-340 was grown on a cell wall preparation from L. sulphureus, the maximal enzyme productivity obtained after 3 days of cultivation was 0.71 U/ml. This yield was about 1.8-fold higher than that achieved on mutan, known so far as the best, but expensive and inaccessible, inducer of mutanase production. Cell-wall-induced mutanase showed a high hydrolytic potential in reaction with a dextranase-pretreated mutan, where maximal degrees of saccharification and solubilization of this biopolymer (80% and 100%, respectively) were reached in 3 h at 45$^{\circ}C$. The mutanase preparation was also effective in degradation of streptococcal mutan and its removal from oral biofilms, especially in a mixture with dextranase.

Toxicity and Changes in Hepatic Metabolizing Enzyme System Induced by Repeated Administration of Pectenotoxin 2 Isolated from Marine Sponges (해면체에서 추출한 Pectenotoxin 2의 마우스에서의 반복적인 투여에 의한 독성 및 간대사효소계에 주는 영향)

  • Yoon, Mi-Young;Kim, Young-Chul
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.280-285
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    • 1997
  • Pectenotoxin 2 (PTX2), isolated from marine sponges, was examined for its hepatotoxic potential using male ICR mice. PTX2 $(20\;or\;100\;{\mu}g/kg/day,\;ip)$ was administered to mice repeatedly for one or two week. Histopathological examination revealed an increase in granularity in the liver from the mice treated with PTX2. PTX2 did not alter the parameters for hepatotoxicity and nephrotoxicity such as sorbitol dehydrogenase (SDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and blood urea nitrogen (BUN). Cytochrome P-450, cytochrome $b_5$, or NADPH cytochrome c reductase was net changed by repeated administration of PTX2. Hepatic microsomal activity of p-nitroanisole O-demethylase, but not aminopyrine N-demethylase, was slightly depressed by PTX2 administerd repeatedly $(100\;{\mu}g/kg/day,\;ip)$ fur 2 weeks. The toxicity of PTX2 $(200\;{\mu}g/kg/day,\;ip)$ was determined in mice pretreated with a metabolic inducer or inhibitor such as phenobarbital, 3-methyl-cholanthrene, $CoCl_2$, or SKF 525-A. Significant alterations in lethality and hepatotoxicity of PTX2 were observed in mice pretreated with a metabolic modulator. The results suggest that liver seems to be the target organ for PTX2 toxicity and also that induction of the PTX2 toxicity may be associated with hepatic drug metabolizing activity.

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Quinone Reductase Inducer from Radish Leaf Cultivated in the Soil Containing Sulfur (유황시비처리가 열무의 Quinone Reductase 유도물질 생성에 미치는 영향)

  • 김경아;노치웅;최경락;황해준;최혜선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.946-950
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    • 2004
  • Young radishes which were grown in the soil containing sulfur increased quinine reductase (QR) activity in Hepa 1clc cells and isothiocyanate-like compound analyzed by HPLC. QR inducing activity was maximum in young radishes grown with 1,818 g/㎥ sulfur and was decreased when the soil was neutralized with lime mortar in order to improve a recovery. These results have suggested that consumption of young radishes, especially grown in the presence of sulfur, would prevent from cancer incidence through inducing detoxification enzymes and could have therapeutic effects for chemoprevention.

Inhibitory effects of artemether on collagen-induced platelet aggregation via regulation of phosphoprotein inducing PI3K/Akt and MAPK

  • Lee, Dong-Ha
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.167-172
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    • 2022
  • Pathophysiological reaction of platelets in the blood vessel is an indispensable part of thrombosis and cardiovascular disease, which is the most common cause of death in the world. In this study, we performed in vitro assays to evaluate antiplatelet activity of artemether in human platelets and attempted to identify the mechanism responsible for protein phosphorylation. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artemether was clarified. Artemether inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artemether decreased TXA2 production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artemether strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC50 of 157.92 μM. These results suggest that artemether has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.