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ABSTRACT: In the course of screening for neuroprotective natural products, Magnoliae Cortex showed potent inhibition of hippocampal neuronal HT22 cell death. Obovatol, honokiol, and magnolol were isolated from the ethanolic extract of Magnoliae Cortex. Isolated compounds obovatol, honokiol, and magnolol were protective against 5 mM glutamate-induced cell death. When cells were stressed using glutamate, cell viability decreased to 16.98 ± 4.58% over the control (100.00 ± 10.15%). In contrast, 10 μM obovatol, 10 μM honokiol, and 50 μM magnolol increased cell viability to 91.80 ± 1.70%, 93.59 ± 1.93%, and 85.36±7.40%, respectively. The neuroprotective effects of obovatol and honokiol were attributable to the inhibition of intracellular reactive oxygen species production, followed by protection of the mitochondrial membrane potential (ΔΨm), recovery of Bcl-2 and Bid levels, inhibition of apoptosis-inducing factor expression, and phosphorylation of mitogen-activated protein kinases such as p38 kinases, extracellular signal-regulated kinases, and c-Jun N-terminal kinases. On the contrary, magnolol did not show any significant effect on the ΔΨm and apoptotic factors. Among three compounds, obovatol most strongly scavenged 2,2-diphenyl-1-picrylhydrazyl radicals and inhibited the elevation of intracellular reactive oxygen species levels in glutamate-stressed HT22 cells. These data suggest that obovatol and honokiol may have clinical applications for preventing neurodegenerative disorders.
Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 02/2013; · 2.99 Impact Factor
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ABSTRACT: High mobility group box 1 (HMGB1) protein is a crucial cytokine that mediates response to infection, injury, and inflammation. Rosmarinic acid (RA) is an important component of the leaves of Perilla frutescens and has neuroprotective, anti-microbial, anti-oxidant, and anti-cancer effects but little is known of its effects on HMGB1-mediated inflammatory response. Here, we investigated this issue by monitoring the effects of RA on the lipopolysaccharide (LPS) or cecal ligation and puncture (CLP)-mediated release of HMGB1 and HMGB1-mediated modulation of inflammatory responses. RA potently inhibited the release of HMGB1 and down-regulated HMGB1-dependent inflammatory responses in human endothelial cells. RA also inhibited HMGB1-mediated hyperpermeability and leukocyte migration in mice. Furthermore, RA reduced CLP-induced HMGB1 release and sepsis-related mortality. Given these results, RA should be viewed as a candidate therapeutic agent for the treatment of various inflammatory diseases via inhibition of the HMGB1 signaling pathway. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.
Journal of Cellular Physiology 10/2012; · 3.87 Impact Factor
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ABSTRACT: Oleanolic acid (OA) is a triterpenoid known for its anti-inflammatory and anti-cancer properties; however, the anti-inflammatory effects of OA on lipopolysaccharide (LPS)-mediated pro-inflammatory responses have not been studied. Here, we first investigated the possible anti-inflammatory effects of OA against pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) induced by LPS and the associated signaling pathways. We found that OA inhibited LPS-induced barrier disruption, expression of cell adhesion molecules (CAMs), and adhesion/transendothelial migration of monocytes to HUVECs. OA also suppressed acetic acid-induced hyperpermeability and carboxymethylcellulose-induced leukocyte migration in vivo. Further studies revealed that OA suppressed the production of tumor necrosis factor-α and activation of nuclear factor-κB by LPS. Collectively, these results suggest that OA has anti-inflammatory effects by inhibiting hyperpermeability, the expression of CAMs, and the adhesion and migration of leukocytes, thereby endorsing its usefulness as a therapeutic agent for vascular inflammatory diseases.
Inflammation 08/2012; · 1.75 Impact Factor
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ABSTRACT: Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but its anticoagulant activities have not been studied. Here, the anticoagulant properties of OA were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrin polymerization as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed OA prolonged aPTT and PT significantly and inhibited thrombin catalyzed fibrin polymerization. In addition, OA inhibited the activities of thrombin and FXa and inhibited the generation of thrombin or FXa in human endothelial cells. OA also inhibited TNF-α-induced tissue factor expression on human endothelial cells. In accordance with these anticoagulant activities, OA showed an anticoagulant effect in vivo. These results indicate that OA possesses antithrombotic activities and suggest that daily consumption of a herb containing OA may be preventing thrombosis in pathological states.
BMB reports 07/2012; 45(7):390-5. · 1.72 Impact Factor
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ABSTRACT: Gynostemma pentaphyllum is widely used in Asian countries as a herbal medicine to treat dyslipidemia, type 2 diabetes and inflammation. An ethanol extract of G. pentaphyllum lessened obesity by activating AMP-activated protein kinase (AMPK). The levels of damulins A and B, components responsible for AMPK activation in the extract, were increased by autoclaving in a time-dependent manner. Heat-processed G. pentaphyllum extract, actiponin containing damulins A (0.93 %, w/w) and B (0.68 %, w/w), significantly stimulated fat oxidation and glucose uptake via AMPK activation in L6 myotube cells. Oral administration of actiponin to ob/ob mice for 8 weeks decreased body weight gain, liver weight, and blood cholesterol levels with AMPK activation in the soleus muscle. Our results demonstrate the beneficial effect of G. pentaphyllum on improving obesity and have elucidated the underlying molecular mechanisms.
Biotechnology Letters 05/2012; 34(9):1607-16. · 1.68 Impact Factor
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ABSTRACT: To analyze the growth inhibitory mechanism of a 2-aminobenzoic acid (2-AA) derived fromBacillus cereus EJ-121, we treatedArabidopsis thaliana plants with 2-AA, 2-AA analogs, auxin (NAA), a known auxin transport inhibitor [2,3,5-triiodobenzoic acid (TIBA)], and an
ethylene action inhibitor [silver thiosulfate (Ag)]. Root development was significantly inhibited by 50 μM 2-AA, whereas the
growth of bacteria and yeast was undeterred. The application of two 2-AA analogs - 3-aminobenzoic acid (3-AA) and 4-aminobenzoic
acid (4-AA) - did not impairArabidopsis root growth at concentrations below 100 μM. These results suggest that the effect of 2-AA is not due to its chemical structure,
but because of its conversion to another metabolite, IAA. To confirm this, we supplemented TIBA in the growth medium, and
found that the degree of inhibition was significantly reduced. Similarly, when plants were co-treated with 100 μM Ag, the
negative effect of 50 μM 2-AA was greatly diminished. All of these observations support the proposal that this inhibition
results from the conversion of 2-AA to IAA. Furthermore, the increased auxin level leads to a rise in ethylene synthesis,
which then blocks root growth and, ultimately, retards overall plant development.
Journal of Plant Biology 05/2012; 50(4):514-516. · 1.07 Impact Factor
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ABSTRACT: This study investigated an ethanol extract from Glycyrrhizae radix (GR), the root of Glycyrrhiza uralensis (Leguminosae), for possible neuroprotective effects on neurotoxicity induced by amyloid β protein (Aβ) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to 10 μM Aβ (25-35) for 36 h induced neuronal apoptotic death. GR (10-50 μg/mL) prevented the Aβ (25-35)-induced neuronal apoptotic death, as assessed by a MTT assay and Hoechst 33342 staining. Furthermore, GR decreased the expression of Bax and active caspase-3, proapoptotic proteins, and increased Bcl-2, an antiapoptotic protein. GR also significantly inhibited Aβ (25-35)-induced elevation of the intracellular Ca(2+) concentration ([Ca(2+)](i)) and generation of reactive oxygen species (ROS) measured by fluorescent dyes. Isoliquiritigenin (1-20 μM), isolated from GR as an active component, inhibited Aβ (25-35)-induced neuronal apoptotic death, elevation of [Ca(2+)](i), ROS generation, and the change of apoptosis-associated proteins in cultured cortical neurons, suggesting that the neuroprotective effect of GR may be, at least partly, attributable to this compound. These results suggest that GR and isoliquiritigenin prevent Aβ (25-35)-induced neuronal apoptotic death by interfering with the increases of [Ca(2+)](i) and ROS, and GR may have a possible therapeutic role for preventing the progression of neurodegenerative disease such as Alzheimer's disease.
Archives of Pharmacal Research 05/2012; 35(5):897-904. · 1.59 Impact Factor
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ABSTRACT: Glycosaminoglycans (GAGs) were isolated from the porcine testis, and their immuno-modulating and anticoagulant activity was
investigated. From anion exchange chromatography (Dowex Macropolous Resin) used for further isolation of porcine testis GAGs
(PT-GAGs), two fractions (PT-GAG-1.5 and PT-GAG-16) eluted by different salt concentration were obtained. In immunomodulating
activity test, PT-GAG-1.5, but not PT-GAG-16, significantly enhanced the growth of murine peritoneal macrophages. In addition,
treatment with PT-GAG-1.5 induced the production of cytokines, interleukin-1β (IL-1β), interferon-γ (IFN-γ) and tumor necrosis
factor-α (TNF-α), from murine microphages. Unexpectedly, both of PT-GAGs had no effect on the growth of murine splenocytes.
The anticoagulant activity of PT-GAG-1.5 and PT-GAG-16 was examined by activated partial thromboplastin time (aPTT) assay
and thrombin time (TT) assay. Both of PT-kGAGs significantly increased the clotting times of aPTT and TT in a dose-dependent
manner. The anticoagulant activity of PT-GAG-16 was found to be higher than that of PT-GAG-1 .5. These results suggest that
PT-GAGs possess biological activities such as immunomodulating activity and anticoagulant activity.
Archives of Pharmacal Research 04/2012; 25(5):669-674. · 1.59 Impact Factor
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ABSTRACT: Anin vitro bioassay-guide revealed that the methanol (MeOH) extract of the stem bark ofPopulus davidiana showed considerable inhibitory activity against cyclooxygenase (COX-1, COX-2). Continuous phytochemical study of the MeOH
extract of this plant led to the isolation of ten flavonoids; sakuranetin (1), rhamnocitrin (2), 7-O-methylaromadendrin (3), naringenin (4), eriodictyol (5), aromadendrin (6), kaempferol (7), neosakuranin (8), sakuranin (9) and sakurenetin-5,4′-di-β-D-glucopyranoside (10). Their structures were identified on the basis of their physicochemical and spectroscopic analyses. The isolated compounds,1–10, were tested for their inhibitory activities against COX-1 and COX-2. Compound7 was found to have potent inhibitory effect on COX-1 and a moderate effect on COX-2, meanwhile, compounds1–6 showed moderate inhibition against COX-1 only. Moreover, compounds5–8 exhibited suppressive effects on xanthine oxidase (XO). These results may explain, in part, the traditional uses ofP. davidiana in ethnomedicine.
Archives of Pharmacal Research 04/2012; 29(12):1102-1108. · 1.59 Impact Factor
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ABSTRACT: Glutamate-mediated excitotoxicity, which is associated with reactive oxygen species (ROS), is hypothesized to be a major contributor to pathological cell death in the mammalian central nervous system, and to be involved in many acute and chronic brain diseases. Here, we showed that isoliquiritigenin (ISL) isolated from Glycyrrhiza uralensis (Gu), one of the most frequently prescribed oriental herbal medicines, protected HT22 hippocampal neuronal cells from glutamate-induced oxidative stress. In addition, we clarified the molecular mechanisms by which it protects against glutamate-induced neuronal cell death. ISL reversed glutamate-induced ROS production and mitochondrial depolarization, as well as glutamate-induced changes in expression of the apoptotic regulators Bcl-2 and Bax. Pretreatment of HT22 cells with ISL suppresses the release of apoptosis-inducing factor from mitochondria into the cytosol. Taken together, our results suggest that ISL may protect against mitochondrial dysfunction by limiting glutamate-induced oxidative stress. In conclusion, our results demonstrated that ISL isolated from Gu has protective effects against glutamate-induced mitochondrial damage and hippocampal neuronal cell death. We expect ISL to be useful in the development of drugs to prevent or treat neurodegenerative diseases.
Biochemical and Biophysical Research Communications 04/2012; 421(4):658-64. · 2.48 Impact Factor
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ABSTRACT: In the course of screening neuraminidase inhibitors from herbal medicines, Reynoutria elliptica exhibited high inhibitory
activity. Four active compounds were isolated from the ethyl acetate soluble fraction by consecutive purification using sillica
gel, Sephadex LH-20 chromatography, and recrystallization. The chemical structures of these compounds were identified as 1,3,8-trihy-droxy-6-methylanthraquinone
(emodin) 1,8-dihydroxy-3-methoxy-6-methylanthraquinone (emodin 3-methyl ether; physcion), 1,3,8-trihydroxy-6-hydoxymethylanthraquinone
(ω-hydroxyemodin), and 3,5,4′-trihydroxystilbene (trans-resvertrol) by spectral data including MS,1 H-, and13 C-NMR. The IC50 values of emodin, emodin 3-methyl ether, ω-hydroxyemodin, andtrans-resvertrol were 2.81, 74.07, 10.49, and 8.77 μM, respectively. They did not inhibit other glycosidase such as glucosidase,
mannosidase, and galactosidase, indicating that they were relatively specific inhibitors of neuraminidase.
Archives of Pharmacal Research 04/2012; 26(5):367-374. · 1.59 Impact Factor
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ABSTRACT: Three diterpenes (1, 8, and9), three triterpenes (3, 4, and7), one saponin (11), four sterols (2, 5, 6, and12), and one cerebroside (10) were isolated from the EtOH extract of the aerial parts ofAralia cordata by repeated silica gel column chromatography. Their chemical structures were identified by comparing their physicochemical
and spectral data with those published in literatures. All isolated compounds were evaluated for their cytotoxicity against
L 1210, K562, and LLC tumor cell lines using MTT assay. Of which, 3β, 5α-dihydroxy-6β-methoxyergosta-7,22-diene (6) showed a potent cytotoxicity against all cell lines with IC50 values of 11.7, 11.9, and 15.1 μM, respectively, while compounds1, 5, and11 showed a moderate or weak cytotoxicity. These isolates were also examined for their inhibitory activity against COX-1 and
COX-2. Although most compounds, except for2, 10, and12, showed a strong inhibitory activity against COX-1, they exhibited a moderate or weak inhibitory activity against COX-2.
Archives of Pharmacal Research 04/2012; 29(7):548-555. · 1.59 Impact Factor
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ABSTRACT: A prolyl endopeptidase inhibitor was isolated from the ethyl acetate soluble fraction ofPhyllanthus ussurensis. The active compound was identified as an ellagitannin, corilagin. It was shown to non-competitively inhibit prolyl endopeptidase
(PEP) with the IC50 value of 1.17x10-6 μM. TheKi value was 6.70x10-7 M. Corilagin was less inhibitory to other serine proteases such as chymotrypsin, trypsin, and elastase, indicating that it
was relatively a specific inhibitor of PEP. Corilagin also effectively inhibited reactive oxygen species such as hydroxide
and superoxide anion radical, hydrogen peroxide, and DPPH. Especially, corilagin showed potent scavenging activity on the
superoxide anion radical in the ESR method (IC50 = 3.79x10-6 M) as well as xanthine oxidase system.
Archives of Pharmacal Research 04/2012; 26(12):1024-1028. · 1.59 Impact Factor
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ABSTRACT: Cytotoxic activity of seven hederagenin saponins isolated from the root ofDipsacus asper were investigatedin vitro against L1210, HL-60, and SK-OV-3 tumor cell lines by the MTT method. 3-O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin (2), 3-O-β-D-xylopyranosyl-(1→3)-α-L-Rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin (6) and 3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl hederagenin (7) exhibited the potent cytotoxicity against the three tumor cell lines with IC50 values ranging from 4.7 to 8.7 μg/mL, with the exception of compound7, which exhibited weak cytotoxic activity against SK-OV-3 (IC50 22.5 μg/mL). Other compounds did not exhibit any cytotoxic activity (IC50>30 μg/mL).
Archives of Pharmacal Research 04/2012; 28(9):1053-1056. · 1.59 Impact Factor
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ABSTRACT: In the course of screening for reactive oxygen species scavengers from natural products, an antioxidant was isolated from
the mycelial culture broth ofPhellinus linteus and identified as hispidin. The hispidin content was reached its maximum level at 12 days after onset of inoculation. About
2.5 mg/mL of hispidin was produced byP. linteus in a yeast-malt medium (pH 5.8, 25°C). Hispidin inhibited 22.6 and 56.8% of the super oxide anion radical, 79.4 and 95.3%
of the hydroxyl radical, and 28.1 and 85.5% of the DPPH radical at 0.1 and 1.0 mM, respectively. The positive control oc-tocopherol
scavenged 25.6 and 60.3%, 74.6 and 96.3%, and 32.7 and 77.5% of each radical, respectively, at the same concentrations. However,
hispidin showed no significant activity on the hydrogen peroxide radical.
Archives of Pharmacal Research 04/2012; 27(6):615-618. · 1.59 Impact Factor
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ABSTRACT: Yellow onion (Allium cepa) extract showed enhanced antioxidative effects in 2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC) and 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate and acetyl ester (CM-H(2)DCFDA) assay after being treated with a crude enzyme extract from soybean paste fungi, Aspergillus kawachii. HPLC analysis showed two increased and two decreased peaks after enzyme treatment. The decreased peaks were identified as quercetin-3,4'-di-O-β-d-glucoside (1) and quercetin-4'-O-β-d-glucoside (2), and peaks that increased were quercetin-3-O-β-d-glucoside (3) and quercetin (4), respectively. It was expected that 3 and 4 were originated from the glucosidic cleavage of their glucosides, 1 and 2. Among the increased compounds, only quercetin (4) showed strong antioxidative activity in the DPPH assay. In addition, the protective effect against glutamate-induced neurotoxicity in HT22 cells was increased when treated with 25 μg/ml of fermented onion. The enhanced neuroprotective effect was also originated from the increased quercetin content. As a consequence, fermentation raised the quercetin content in onion, and subsequently increased the antioxidative and neuroprotective activities.
Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 04/2012; 50(6):2042-8. · 2.99 Impact Factor
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ABSTRACT: As a late mediator of inflammation, high mobility group box 1 (HMGB1) protein up-regulates pro-inflammatory cytokines in several inflammatory diseases. Further, high plasma levels of HMGB1 correlate with poor prognosis and increased mortality in patients with severe inflammation. Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but the effects of OA on HMGB1-mediated pro-inflammatory responses of human endothelial cells is not well-studied. In this study, we investigated this question by monitoring the effect of OA on lipopolysaccharide (LPS)-mediated release of HMGB1 and the HMGB1-mediated modulation of inflammatory responses in human umbilical vein endothelial cells (HUVECs). OA potently inhibited the release of HMGB1 by HUVECs as well as down-regulated HMGB1-dependent adhesion and migration of the monocytic cell line THP-1 to activated HUVECs. OA also down-regulated the cell surface expression of the receptor of HMGB1, thereby inhibiting HMGB1-dependent pro-inflammatory responses by inhibiting activation of nuclear factor-κB (NF-κB) and production of tumor necrosis factor-α (TNF-α) by HMGB1. Given these results, OA showed anti-inflammatory activities and could be a candidate as a therapeutic agent for various inflammatory diseases through the inhibition of the HMGB1 signaling pathway.
Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 02/2012; 50(5):1288-94. · 2.99 Impact Factor
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ABSTRACT: This study investigated a methanol extract from the leaf and stem of Vitis amurensis (Vitaceae) for possible neuroprotective effects on neurotoxicity induced by amyloid β protein (Aβ) (25-35) in cultured rat cortical neurons and also for antidementia activity in mice. Exposure of cultured cortical neurons to 10 μM Aβ (25-35) for 36 h induced neuronal apoptotic death. At concentrations of 1-10 μg/mL, V. amurensis inhibited neuronal death, the elevation of intracellular calcium ([Ca(2+)](i)) and the generation of reactive oxygen species (ROS), all of which were induced by Aβ (25-35) in primary cultures of rat cortical neurons. Memory loss induced by intracerebroventricular injection of ICR mice with 16 nmol Aβ (25-35) was inhibited by chronic treatment with V. amurensis extract (50 and 100 mg/kg, p.o. for 7 days), as measured by a passive avoidance test. Amurensin G, r-2-viniferin and trans-ɛ-viniferin isolated from V. amurensis also inhibited neuronal death, the elevation of [Ca(2+)](i) and the generation of ROS induced by Aβ (25-35) in cultured rat cortical neurons. These results suggest that the neuroprotective effect of V. amurensis may be partially attributable to these compounds. These results suggest that the antidementia effect of V. amurensis is due to its neuroprotective effect against Aβ (25-35)-induced neurotoxicity and that the leaf and stem of V. amurensis have possible therapeutic roles for preventing the progression of Alzheimer's disease.
Archives of Pharmacal Research 10/2010; 33(10):1655-64. · 1.59 Impact Factor
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ABSTRACT: The medicinal herb Jinpi, derived from the dried stem barks of Fraxinus rhynchophylla belonging to Oleaceae is widely used as a variety of Korean folk remedies for anti-inflammatory, febricide, antidiarrhea, and antileukorrhea diseases. In the course of screening antidementia agents from natural products, F. rhynchophylla showed significant inhibitory activity toward Abeta(25-35)-induced neuronal cell death. An active principle was isolated and identified as syringin. When the neuroblastoma cells were exposed to 50 microM Abeta(25-35), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction rate (survival rate) decreased to 60.21 +/- 2.16% over control while syringin treated ones recovered cell viability up to 79.12 +/- 1.39% at 20 microM. In addition, 20 microM syringin almost completely removed Abeta(25-35)-induced reactive oxygen species. The neuroprotective effect of syringin seemed to be originated from the reduction of apoptosis since decrease in caspase-3 activity and expression, reduction in cleaved PARP, and DNA fragmentation were observed. These results suggest that F. rhynchophylla and syringin are expected to be useful for preventing Abeta(25-35)-induced neuronal cell damage.
Archives of Pharmacal Research 04/2010; 33(4):531-8. · 1.59 Impact Factor
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ABSTRACT: Amylases have significant importance in broad industrial application including bio-ethanol production. Although amylases are widely distributed in microbes, plants and animals, it has been sought for new amylases from various sources with special industrial potential. In this study we firstly isolated and characterized a novel thermostable alpha-amylase from Korean pine seed. Enzyme was purified to homogeneity level with purification fold of 1286.1 using several techniques such as self-precipitation, (NH(4))(2)SO(4) fractionation, DEAE anion exchange and starch affinity chromatography. The purified alpha-amylase showed two bands in SDS-PAGE with molecular weight of 44 and 45 kDa. The apparent molecular weight of native enzyme was calculated to be 46.7 kDa. Internal peptide sequencing confirmed that the purified alpha-amylase was a novel enzyme. The optimum pH and temperature for enzyme activity were pH 4.5 and 65 degrees C, respectively. This enzyme was fully stable for 48h at 50 degrees C and retained 80% activity up to 96h. The K(m) and V(max) were 0.84 mg/ml and 3.71 micromol/min, respectively. On the basis of high thermal stability and a broad range of pH stability, the pine seed alpha-amylase showed a good prospect of industrial application.
New Biotechnology 09/2009; 26(3-4):143-9. · 2.76 Impact Factor
