• Title/Summary/Keyword: cell-free extract

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Reduction of hexavalent chromium by pseudomonas aeruginosa HP014

  • Oh, Young-Sook;Choi, Sung-Chan
    • Journal of Microbiology
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    • 제35권1호
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    • pp.25-29
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    • 1997
  • Microbial reduction of hexavalent (VI) to trivalent (III) chromium decreases its toxicity by two orders of magnitude. In order to investigate the nature of Cr-reduction, Cr-resistant Pseudomonas aeruginosa HP014 was isolated and tested for its reduction capability. At the concentration of 0.5 mM Cr(VI), cell growth was not inhibited by the presence of Cr(VI) in a liquid medium, and Cr(VI) reduction was accompanied by ell growth. When cell-free extract was tested, the reduction of Cr(VI) showed a saturation kinetics with the maximum specific activity of 0.33 .mu.mol min$\^$-1/ mg$\^$-1/ cell protein, and an apparent K. of 1.73 mM Cr(VI). The activity required either NADH or NADPH as an electron donor. However, NADPH gave 50% as mush activity as sequently the supernatant and pelleted membrane fractions were tested for Cr(VI) reduction activity. The supernatant of the centrifugation showed almost the same Cr(VI) reduction activity as compared with that of the cell-free extract, indicating that the Cr(VI)-reducing activity of P. aeruginosa HP-14 is due to soluble enzyme. Moreover, the activity appeared to be the highest among the known activities, suggesting that the strain might be useful for remediation of Cr(VI)-contaminated sites.

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연자육이 음경해면체 이완에 미치는 영향 (Relaxation Effects of Nelumbinis Semen in Isolated Corpus Cavernosum)

  • 박선영;김진택;김호현
    • 동의생리병리학회지
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    • 제29권2호
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    • pp.180-188
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    • 2015
  • This study was conducted to investigate the relaxation effects and its mechanisms of Nelumbinis Semen(NS) extract in isolated rabbit corpus cavernous tissues. In order to examine the relaxation effects and its mechanisms of NS, we treated the ethanol extract of NS(0.01-3.0 mg/ml) and indomethacin(IM), tetraethylammonium chloride(TEA), Nω -nitro-L-arginine (L-NNA), methylene blue(MB) were treated before NS extract to contracted strips induced by PE 1 μM. We also treated calcium chloride(Ca) 1 mM after pretreatment of NS extract in Ca2+-free krebs-ringer solution to contracted strips induced by PE. Cell viability and NO concentration on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, Griess reagent system. eNOS production was investigated by histochemical and immunohistochemical staining. NS extract was significantly affected on the relaxation of cavernous strips and NS extract-induced relaxation was not different by pretreatment of IM, TEA, MB, but inhibited by the pretreatment of L-NNA. And increase of contraction induced by Ca2+ addition, in a Ca2+-free solution, was decreased by pretreatment of NS. NO concentration on HUVEC was increased. When NS extract was applicated on corpus cavernosum of penis(CCP) in SHR, ratio of smooth muscles to collage fibers by PE was decreased and formation of eNOS around helicine artery was increased. These results suggest that CCP relaxation effects of NS extract are shown by suppressing influx of extracellular Ca2+ through the production of NO and eNOS.

소합향원(蘇合香元)이 혈액내 Free Cholesterol에 의한 혈관내피세포 손상에 미치는 영향 (The Effect of Sohaphyangwon water extract on Endothelial cells by Free choleserol in blood)

  • 이상헌;이소연;윤현덕;신오철;박창국;박치상
    • 대한한방내과학회지
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    • 제26권3호
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    • pp.575-586
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    • 2005
  • FC(free choresterol) plays an important role in normal and pathophysiological cells including that of messenger molecule or dilator of blood vessels in such illnesses as artheriosclerosis, hypertension and myocardial infarction. Smooth muscle and endothelial cell functions in the arteria wall are unified by complex intercellular signalling processes. In arteria comprised of one layer of smooth muscle cells surrounding the endothelium, the close apposition of the two cell types enables a signal derived from one cell to rapidly diffuse to neighboring cells. Experimentation was conducted to investigate the potential contribution of Sohaphyangwon(SHHW) on levels of FC generated by goaded microphages, and mechanisms of protection against ACAT inhibitor. It was found that J774 macrophages, which normally do not express FC were expressed by oxLDL and ACAT inhibitor. SHHW protected cells were found to be resistant to oxLDL and delayed death following the FC. Inhibition of FC formation abolished the protective effect against ACAT inhibitor exposure. Cadiovascular diseases include abnormalities of blood vessels dysfunction of the renin-angiotension system. What relation herbal medicine may have with vessel endothelium necrosis was here studied. In Oriental Medicine, SHHW water extract used for diseases in relation to cardiovascular systems. The resistence to cardiovascular disease of ACAT inhibitor induced J774 macrophage cells were studied through analysis of cell morphological patterns and immunochemistry of SHHW. The results of this study suggest that SHHW has protective effects on the cardiovascular system, and that it is effective in both prevention and treatment of diseases of the cardiovascular system, particularity against necrosis of blood.

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Enantioselective N-Acetylation of 3-Amino-3-phenylpropionic Acid by Cell-free Extracts of Streptomyces neyagawaensis

  • Chung, Myung-Chul;Lee, Ho-Jae;Lee, Choong-Hwan;Chun, Hyo-Kon;Kho, Yung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제7권5호
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    • pp.329-332
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    • 1997
  • Cell-free extracts of Streptomyces neyagawaensis SL-387 grown on a chemically defined medium supplemented with DL-3-amino-3-phenylpropionic acid (APP) produced N-acetyl-APP (Ac-APP) in the presence of APP and acetyl coenzyme A. The APP obtained by acid hydrolysis of the Ac-APP was D-configuration: $[\alpha]_D+6.5^{\circ}(H_2O)\;at\;20^{\circ}C$, optical purity 92% enantiomeric excesses (ee). These results suggest that an N-acetyltransferase exists in the cell-free extract as a novel enzyme with specificity for D-APP.

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Continuous Cell-Free Protein Synthesis Using Glycolytic Intermediates as Energy Sources

  • Kim, Ho-Cheol;Kim, Tae-Wan;Park, Chang-Gil;Oh, In-Seok;Park, Kyung-Moon;Kim, Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.885-888
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    • 2008
  • In this work, we demonstrate that glycolytic intermediates can serve as efficient energy sources to regenerate ATP during continuous-exchange cell-free (CECF) protein synthesis reactions. Through the use of an optimal energy source, approximately 10 mg/ml of protein was generated from a CECF protein synthesis reaction at greatly reduced reagent costs. Compared with the conventional reactions utilizing phosphoenol pyruvate as an energy source, the described method yields 10-fold higher productivity per unit reagent cost, making the techniques of CECF protein synthesis a more realistic alternative for rapid protein production.

흰털오가피 부위별 물추출물의 항산화활성 (Antioxidative Activity of Water Extract of Different Parts of Acanthopanax divaricatus var, albeofructus)

  • 유수연;김지영;노빛나;박원봉
    • 약학회지
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    • 제50권3호
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    • pp.191-198
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    • 2006
  • Acanthopanax species have traditionally been used as a tonic, a sedative as well as in the treatment of rheumatism, hypertension and diabetes. In the present study, oxidative stress was induced in Vero cells by incubating the cells with glucose and the cell viability was measured by MTT assay. The concentration of glucose which 50% of cell viability was 125 mM $(IC_{50})$ and the cell viability was increased to $87.6{\pm}8.8%$ by treatment of the extracts of Acanthopanax divaricatus var. albeofructus. The antioxidative activity of water extract of different parts of the Acanthopanax plant was investigated by DPPH (1,1-diphenyl-2-picrylhydrazyl) assay, xylenol orange assay, TBARS (thiobarbituric acid reactive substances) assay and enzyme (superoxide anion and catalase) assay. Each extract (leaves, root, stem and fruits) of the plant showed free radical and $H_2O_2$ scavenging activity. The extract also inhibited lipid peroxidation and recovered enzyme (superoxide anion dismutase and catalase) activity in Vero cells treated with glucose.

Inhibitory effects of the extract of Viscum album on the proliferation of human tumor cell lines

  • Seo, Jee-Hee;Choi, Yeon-Hee;Kim, Jung-Sook;Kim, Seong-Kie;Choi, Sang-Un;Kim, Young-Sup;Ryu, Shi-Yong;Kim, Young-Kyoon
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.202.2-202.2
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    • 2003
  • A bioassay-guided fractionation of the whole extract of Viscum album (a parasitic plant : Loranthaceae) led to the isolation of two triterpenoidal components, oleanolic acid (1), ${\beta}$-amyrin acetate (2), homoflavoyadorinin B (3) as well as large quantity of free fatty acid mixtures as active ingredients of the extract responsible for the antitumoral property. The EtOAc soluble part and BuOH soluble part of the extract demonstrated a significant inhibition on the proliferation of cultured human tumor cells such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT-15 (colon) in vitro, whereas the remaining water soluble part exhibited a poor inhibition. (omitted)

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Aspergillus oryzae에 있어서 L-Tyrosine의 분해효소에 관한 연구 (Studies on the Degradation of L-Tyrosine by Aspergillus oryzae)

  • 정동효;박성오;김영진
    • Applied Biological Chemistry
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    • 제14권2호
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    • pp.131-135
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    • 1971
  • 1. Aspergillus oryzae의 균체에는 L-tyrosine-${\alpha}$-ketoglutaric acid transaminase와 p-hydroxyphenlypyruvic acid oxidase가 존재해 있다. 2. L-Tyrosine 산화효소는 액침배양한 Aspergillus oryzae의 acetone powder, cell free extract 및 배양액에도 존재하며 L-tyrosine은 ${\alpha}$-ketoglutaric acid의 첨가로 더욱 빨리 전환되었다. 3. ${\alpha}$-Ketoglutaric acid와 pyridoxal phosphate는 transamination의 amino기의 수용체로 생각되었다. 4. 이들 효소계는 L-tyrosine와 p-hydroxyphenlypyruvic acid를 homogentisic acid로 산화시켰다. 5. Ascorbic acid는 p-hydroxyphenlypyruvic가 homogentisic acid로 산화되는데 특별한 역할을 하는 것 같다. 6. L-Tyrosine-${\alpha}$-ketoglutaric acid transaminase와 p-hydroxyphenlypyruvic acid oxidase의 최적 pH는 각 각 pH $6.0{\sim}6.5$와 pH 7.5이었다.

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무막줄기세포추출물의 3T3-L1 세포에서 포도당 흡수 촉진 효과 (Membrane Free Stem Cell Extract from Adipose Tissue Enhances Glucose Uptake in 3T3-L1 Cells)

  • 김지현;김민정;박혜숙;김영실;조은주
    • 한방비만학회지
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    • 제19권2호
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    • pp.89-96
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    • 2019
  • Objectives: We investigated whether membrane free stem cell extract from adipose tissue (MFSCE) has anti-diabetic effect. Methods: To determine glucose uptake effect of MFSCE, we carried out glucose uptake assay in 3T3-L1 adipocytes. The regulatory mechanisms of MFSCE on glucose uptake were examined by Western blot analysis. Results: When MFSCE was treated to adipocytes at the concentration of 0.5, 1, 2.5, and 5 ㎍/mL, 2-deoxyglucose-6-phosphate uptake was elevated approximately 1.8-fold compared to cells not treated with MFSCE. It indicated that MFSCE enhances glucose uptake in 3T3-L1 adipocytes. In addition, MFSCE reduced phosphorylation of insulin receptor substrate-1 at serine 307 and induced Akt and glucose transporter 4 protein expressions that were related to insulin signaling. Furthermore, MFSCE regulated adenosine monophosphate-activated protein kinase (AMPK) pathway by increases of increase phosphorylation of AMPK and acetyl-CoA carboxylase that were related to AMPK pathway. Conclusions: These results indicated that MFSCE promotes glucose uptake via modulation of insulin signaling and AMPK pathway. Therefore, MFSCE could be a promising agent for treatment of diabetes mellitus.

시호사물탕이 t-BHP로 유도된 간세포의 산화적 손상 및 자유기에 의한 지질과산화 반응에 미치는 영향 (Preventive Effects of Sihosamultang Extract on Lipid Peroxidation by Free Radicals and Oxidative Damage of Hepatocytes by tert-Butyl Hydroperoxide)

  • 김태기;박선동;문진영
    • 대한한의학방제학회지
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    • 제8권1호
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    • pp.241-255
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    • 2000
  • Sihosamultang(SST) has been used for the treatment of puerperal fever, liver disease in traditional medicine. The present study was carried out to evaluate the antioxidant effects of SST extract in vitro. The inhibitory effect of SST extract on lipid peroxidant was examined in the linoleic acid autoxidation system. In this test, SST extract significantly inhibited the time course of the lipid peroxidation. And SST extract showed about 73% scavenging effect on DPPH radical. And this extract inhibited not only the lipid peroxide formation induced by hydroxyl radical derived from $ H_{2}O_{2}-Fe^{2+}$ in the rat liver homogenate, but also the superoxide generation from xanthine-xanthine oxidase system in a dose-dependent manner. In addition, SST extract protected the hepatic cell death induced by tert-butyl hydroperoxide. These data indicated that SST might play a protective role against oxidative injury by free radicals.

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