• Title/Summary/Keyword: L-ribose

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Evaluation on Anticancer Effect Against HL-60 Cells and Toxicity in vitro and in vivo of the Phenethyl Acetate Isolated from a Marine Bacterium Streptomyces griseus

  • Lee, Ji-Hyeok;Zhang, Chao;Ko, Ju-Young;Lee, Jung-Suck;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.18 no.1
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    • pp.35-44
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    • 2015
  • We previously identified Streptomyces griseus as an anti-cancer agent (Kim et al., 2014). In this study, we isolated compounds from S. griseus and evaluated their anticancer effect and toxicity in vitro and in vivo. Preparative centrifugal partition chromatography (CPC) was used to obtain three compounds, cyclo($_{\small{L}}$-[4-hydroxyprolinyl]-$_{\small{L}}$-leucine], cyclo($_{\small{L}}$-Phe-trans-4-hydroxy-$_{\small{L}}$-Pro) and phenethyl acetate (PA). We chose PA, which had the highest anticancer activity, as a target compound for further experiments. PA induced the formation of apoptotic bodies, DNA fragmentation, DNA accumulation in $G_0/G_1$ phase, and reactive oxygen species (ROS) formation. Furthermore, PA treatment increased Bax/Bcl-xL expression, activated caspase-3, and cleaved poly-ADP-ribose polymerase (PARP) in HL-60 cells. Simultaneous evaluation in vitro and in vivo, revealed that PA exhibited no toxicity in Vero cells and zebrafish embryos. We revealed, for the first time, that PA generates ROS, and that this ROS accumulation induced the Bcl signaling pathway.

Purification and Characterization of the D-xylulokinase from Candida sp. L-16 (Candida sp. L-16이 생산하는 D-Xylulokinase의 정제 및 특성)

  • 이종수;주길재
    • Food Science and Preservation
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    • v.9 no.4
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    • pp.429-433
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    • 2002
  • The D-xylulokinase from Candida sp. L-16 was purified through a sequence of ammonium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-100 and Sephadex G-200 gel filtration. The specific activity of the purified Dxylulokinase was increased to 23.2 fold and the yield was 11.2%. The enzyme was showed to be a single protein band by SDS-PAGE. The molecular weight of the enzyme was 150,000 dalton, this enzyme was identified to be a dimer with two subunits. The optimum conditions of the enzyme were pH 8.0 and 40$\^{C}$, respectively. The enzyme was relatively stable between pH 7.0 to pH 9.0, but it was unstable over 30$\^{C}$. The enzyme showed substrate specificity on D-xylulose, D-arabinose and D-ribose, Km value and Vmax for D-xylulose were 0.042 mM and 117 units/ml, respectively. The activation energy of the enzyme was 4.75 Kcal/mol. The one was inhibited by metabolic intermediates such as 6-phosphogluconic acid, 2-keto-gluconic acid. The enzyme was activated by EDTA and thiol compounds such as cysteine-HCI, DTT and glutathione.

Comparative analysis of total sugar and sugar alcohol contents of the fruiting bodies in edible and medicinal mushrooms (식용 및 약용버섯류의 단당류, 이당류, 당알코올 함량 비교 분석)

  • Cho, Jae-Han;Park, Hye-Sung;Han, Jae-Gu;Lee, Gang-Hyo;Sung, Gi-Ho;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.12 no.4
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    • pp.316-323
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    • 2014
  • We analyzed saccharide by dividing and comparing Monosaccharide, Disaccharide and sugar Alcohol. At first, Glucose had outstanding contained quantity of ASI 7114 with 81.11 g/l even comparing with other mushrooms for medical use and edibility. And 119.98 g/l of Fructose was observed at Hericium erinaceum that was more contained quantity than Flammulina velutipes and Lentinus edodes. But, the most contained quantity observed in Ganoderma lucidum was ASI 7015 with 15.70 g/l that was the level of 1/8 approximately against Hericium erinaceum. Ribose was found at low level generally that was hardly contained. Xylose was also observed low level. ASI 7004 was detected at 0.96g/l that was the most content with imperceptible difference by comparing with other mushrooms for medical use and edibility. Next, 35.21 g/l of Trehalose, disaccharide was observed at Agaricus bisporus that was around 11 times of content than ASI 3.09 g/l that was the most content of Ganoderma lucidum. For ${\alpha}$-Lactose, Sparassis crispa has the most amount of 3.38 g/l that was around 12.5 times of ASI 7060 0.27 g/l that was the most content of Ganoderma lucidum. For Glycerol, sugar alcohol, 64.74 g/l was observed at Pleurotus eryngii. We knew it was around 8 times of ASI 7004 8.61 g/l that was the most content of Ganoderma lucidum. 0.72 g/l of Solbitol was observed at Flammulina velutipes. We knew it was around 2times of ASI 7003 0.31 g/l that was the most content of Ganoderma lucidum. Moreover most of Ganoderma lucidum didn't contain Solbitol. 2.96 g/l of Mannitol was observed at Agaricus bisporus. that was the most content among other mushrooms. Also Mannitol was contained in Lentinus edodes and leurotus cornucopiae only. Even Ganoderma lucidum didn't have Mannitol. At last, as a result of myo-Inosito content analysis, it was seemed not to be involved in any of mushrooms.

In Vitro Antiproliferative and Apoptosis Inducing Activity of Bibimbap on HT-29 Human Colorectal Adenocarcinoma Cells

  • Tae Eun, Guon;Ha Sook, Chung
    • The Korean Journal of Food And Nutrition
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    • v.35 no.6
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    • pp.464-472
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    • 2022
  • The present study was designed to investigate the antiproliferative activity and molecular mechanisms of Bibimbap in HT-29 human colorectal adenocarcinoma cells. Bibimbap extract inhibited the proliferation of HT-29 cells by 50% at a concentration of 10.1±0.17 mg/mL for 48 h. The population of live cells decreased slightly, and the morphology changed with a reduction in cell volume (pyknosis) with Bibimbap. Treatment with 5 mg/mL of Bibimbap resulted in slight cell shrinkage. Furthermore, as the Bibimbap dose increased to 10 mg/mL, these characteristics were more evident, and HT-29 cells exhibited partial detachment by staining with the DNA-binding dye Hoechst 33342. Flow cytometric analysis by Annexin V and PI double staining showed that Bibimbap increased the levels of apoptosis. Analysis of the mechanism of these events showed that Bibimbap-treated cells exhibited a mitochondria-dependent apoptotic pathway through the modulation of caspase-3, caspase-8, caspase-9, and poly-ADP ribose polymerase, as well as Bax and Bcl-2 expression in dose- and time-dependent manners. Consequently, Bibimbap exerts a significant antiproliferative effect on HT-29 human colorectal adenocarcinoma cells.

Biological Active Substance Produced by a Strain of Streptomyces sp. (Part.III) Purification and Nutritional Requirement. (Streptomyces 속 균주가 생성한 물질의 생물활성 (제삼보) 정제 및 영양요구성)

  • 송방호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.5 no.1
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    • pp.36-45
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    • 1977
  • A piscicidal substance was isolated from the culture medium of Streptomyces umbrosus by avicel column chromatography and avicel thin layer chromatography after extration with chroloform. Bluegreen fluorescence was emitted under UV irradiation. Factors which govern toxin production and nutrition requirement for high toxin titres were observed. Nutritional uptake for toxin production was not curresponded with that for cell growth. Alanine, valine, serine asparagine, arginine, histidine, urea and sodium nitrate as a carbon source and glucose, mannose, rhamnose, xylose, arabitol and starch as a carbon source were recognized as a favorable nutrient for high toxin production. Magnesium was essential factor whereas vitamins were not of effective. Most of toxin was formed simultaneously with cell growth in esponential phase. Maximal production was observed for six day culture at 3$0^{\circ}C$. Tissues of gill, kidney and pnacreas in Cyprinus carpio were denatured extreamly after treating with the substance. Atrophied nucleous, indented membrane and degradated cytoplasm with necrotic affectness were noted on each tissue. The chemical formula of the substance was designated as $C_{38}$ $H_{66}$ $NO_4$.

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Effects of Pentoses on 2-deoxy-D-Glucose Transport of the Endogenous Sugar Transport Systems in Spodoptera frugiperda Clone 9 Cells

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.55-60
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    • 2009
  • Insect cells such as Spodoptera frugiperda Clone 9 (Sf9) cells are widely chosen as the host for heterologous expression of a mammalian sugar transport protein using the baculovirus expression system. Characterization of the expressed protein is expected to include assay of its function, including its ability to transport sugars and to bind inhibitory ligands such as cytochalasin B. It is therefore very important first to establish the transport characteristics and other properties of the endogenous sugar transport proteins of the host insect cells. However, very little is known of the transport characteristics of Sf9 cells, although their ability to grow on TC-100 medium strongly suggested the presence of endogenous glucose transport system. In order to investigate the substrate and inhibitor recognition properties of the Sf9 cell transporter, the ability of pentoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. To determine the time period over which of sugar into the Sf cells was linear, the uptake of 2dGlc 0.1mM extracellular concentration was measured over periods ranging from 30 seconds to 30 minutes. The uptake was linear for at least 2 minutes at the concentration, implying that uptake made over a 1 minute time course would reflect initial rates of the sugar uptake. The data have also revealed the existence of a saturable transport system for pentose uptake by the insect cells. The transport was inhibited by D-xylose and D-ribose, although not as effective as hexoses. However, L-xylose had a little effect on 2dGlc transport in the Sf9 cells, indicating that the transport is stereoselective. Unlike the human erythrocyte-type glucose transport system, D-ribose had a somewhat greater apparent affinity for the Sf9 cell transporter than D-xylose. It is therefore concluded that Sf9 cells contain an endogenous sugar transport activity that in some aspects resembled the human erythrocyte-type counterpart, although the Sf9 and human transport systems do differ in their affinity for cytochalasin B.

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Involvement of Oxidative Stress and Poly(ADP-ribose) Polymerase Activation in 3-Nitropropionic Acid-induced Cytotoxicity in Human Neuroblastoma Cells

  • Nam, Eun-Joo;Lee, Young-Jae;Oh, Young-Ah;Jung, Jin-Ah;Im, Hye-In;Koh, Seong-Eun;Maeng, Sung-Ho;Joo, Wan-Seok;Kim, Yong-Sik
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.325-331
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    • 2003
  • 3-Nitropropionic acid (3-NP) inhibits electron transport in mitochondria, leading to a metabolic failure. In order to elucidate the mechanism underlying this toxicity, we examined a few biochemical changes possibly involved in the process, such as metabolic inhibition, generation of reactive oxygen species (ROS), DNA strand breakage, and activation of Poly(ADP-ribose) polymerase (PARP). Exposure of SK-N-BE(2)C neuroblastoma cells to 3-NP for 48 h caused actual cell death, while inhibition of mitochondrial function was readily observed when exposed for 24 h to low concentrations (0.2${\sim}$2 mM) of 3-NP. The earliest biochemical change detected with low concentration of 3-NP was an accumulation of ROS (4 h after 3-NP exposure) followed by degradation of DNA. PARP activation by damaged DNA was also detectable, but at a later time. The accumulation of ROS and DNA strand breakage were suppressed by the addition of glutathione or N-acetyl-L-cysteine (NAC), which also partially restored mitochondrial function and cell viability. In addition, inhibition of PARP also reduced the 3-NP-induced DNA strand breakage and cytotoxicity. These results suggest that oxidative stress and activation of PARP are the major factors in 3-NP-induced cytotoxicity, and that the inhibition of these factors may be useful in protecting neuroblastoma cells from 3-NP-induced toxicity.

Characterization and ACE Inhibitory Activity of Fermented Milk with Probiotic Lactobacillus plantarum K25 as Analyzed by GC-MS-Based Metabolomics Approach

  • Zhang, Min;Jiang, Yunyun;Cai, Miao;Yang, Zhennai
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.903-911
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    • 2020
  • Addition of probiotics to yogurt with desired health benefits is gaining increasing attention. To further understand the effect of probiotic Lactobacillus plantarum on the quality and function of fermented milk, probiotic fermented milk (PFM) made with probiotic L. plantarum K25 and yogurt starter (L. delbrueckii ssp. bulgaricus and Streptococcus thermophilus) was compared with the control fermented milk (FM) made with only the yogurt starter. The probiotic strain was shown to survive well with a viable count of 7.1 ± 0.1 log CFU/g in the PFM sample after 21 days of storage at 4℃. The strain was shown to promote formation of volatiles such as acetoin and 2,3-butanediol with milk fragrance, and it did not cause post-acidification during refrigerated storage. Metabolomics analysis by GC-MS datasets coupled with multivariate statistical analysis showed that addition of L. plantarum K25 increased formation of over 20 metabolites detected in fermented milk, among which γ-aminobutyric acid was the most prominent. Together with several other metabolites with relatively high levels in fermented milk such as glyceric acid, malic acid, succinic acid, glycine, alanine, ribose, and 1,3-dihydroxyacetone, they might play important roles in the probiotic function of L. plantarum K25. Further assay of the bioactivity of the PFM sample showed significant (p < 0.05) increase of ACE inhibitory activity from 22.3% at day 1 to 49.3% at day 21 of the refrigerated storage. Therefore, probiotic L. plantarum K25 could be explored for potential application in functional dairy products.

Apicidin-Mediated Apoptosis Signaling in Human Promyelocytic Leukemia U937 Cells (Apicidin, Histone-Deacetylase Inhibitor에 의한 Promyelocytic U937 세포고사)

  • 정은현;박찬희;임창인;이황희;송훈섭;염성섭;정은배;이병곤;김영훈
    • Toxicological Research
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    • v.19 no.3
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    • pp.197-203
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    • 2003
  • Apicidin, a histone-deacetylase inhibitor, has been successfully used to inhibit the growth of cancer cells. In this study, the apoptotic potential and mechanistic insights of apicidin were investigated in human myeloid leukemia U937 cells. Treatment of U937 cells with apicidin resulted in a decrease of cell viability with apoptotic characteristics, including chromatin condensation and ladder-pattern fragmentation of genomic DNA. Apicidin converted the procaspase-3 protease to catalytically active effector protease, resulting in subsequent cleavage of poly (ADP-ribose) polymerase (PARP) and inhibitor of caspase-activated deoxyribonuclease (ICAD). In addition, apicidin induced the activation of caspase-9 protease and the cytosolic release of mitochondrial cytochrome c with mitochon-drial membrane potential transition. Moreover, apicidin transiently increased the expression of Fas and Fas ligand proteins. Taken together, the results suggest that apicidin induces apoptosis of U937 cells through activation of intrinsic caspase cascades and Fas/FasL system with mitochondrial dysfunction.

Induction of Apoptosis in Human Oral Epidermoid Carcinoma Cells by Essential Oil of Chrysanthemum boreale Makino

  • Cha, Jeong-Dan;Jeong, Mi-Ran;Lee, Young-Eun
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.350-354
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    • 2005
  • The effect of the essential oil obtained from Chrysanthemum boreale Makino on the apoptosis of KB cells was investigated. Cytotoxicity and cellular DNA content were analyzed by MTT assay, flow cytometry, agarose gel electrophoresis, and Hoechst 33258 staining. The caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were estimated by Western blotting method. The various cytotoxic effects of the essential oil which are hallmarks of apoptosis, including DNA fragmentation, apoptotic body formation, and sub-G1 DNA content, all progressed in a dose-dependent manner. Treatment with an apoptosis-inducing concentration of the essential oil caused rapid and transient induction of caspase 3 activity. Further, the efficacious induction of PARP cleavage and caspase-3 activation was observed at an essential oil concentration of 0.1 and 0.2 mg/mL for 12 hr.