• Title/Summary/Keyword: Methanol dehydrogenase

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Protective Effects of Houttuynia cordata Thunb on Carbon Tetrachloride-induced Hepatotoxicity in Rats (어성초 분획물이 사염화탄소로 유발된 흰쥐의 간손상에 대한 보호효과)

  • Kim, Ok-Kyung
    • Korean Journal of Pharmacognosy
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    • v.33 no.4 s.131
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    • pp.324-331
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    • 2002
  • This study was performed to investigate the protective effect of Houttuynia cordata Thunb on hepatotoxicity in carbon tetrachloride$(CCI_4)$ intoxicated rats. The examined effects hexane, chloroform, butanol and water fractions prepared from the Houttuynia cordata Thunb methanol exlract and rats were administrated with those orally once a day for successive 6 days, fellowed by treaoent with $CCl_4$ on the sixth day. After 6 days, the activities of aminotransferase, alkalinephosphatase, ${\gamma}-glutamyl$ transpeptidase, lactate dehydrogenase and contents of triglyceride, hepatic lipid peroxide in butanol fraction pretreated rats were significantly decreased compared to the only $CCl_4$ treated rats, also depletion glutathione content induced by treatment with $CCl_4$ was prevented by butanol fraction pretreated rats. In addition, activities of hepatic superoxide dismutase, catalase, glutathione peroxidase in butanol fiaction pretreated rats were significantly decreased compared to the only $CCl_4$ treated rats, but the activity of hepatic glutathione-S-transferase was not significantly effect. These results suggest that butanol fiuction of Houttuynia cordata Thunb methanol extract have potent hepatoprotective effect against carbon tetrachloride intoxicated rats.

Nitric Oxide Production Inhibitory Effect and Antibacterial Activity of the Extract and Fractions from Paeoniae Radix (작약 메탄올 추출물 및 분획물의 Nitric Oxide 생성 억제 효과와 피부질환 원인균에 대한 항균활성)

  • Im, Do-Youn;Lee, Kyoung-In
    • Korean Journal of Pharmacognosy
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    • v.43 no.2
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    • pp.173-178
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    • 2012
  • In this study, we investigated antibacterial activity and nitric oxide production inhibitory effect of the methanol extract and its fractions from Paeoniae Radix. In antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and P. aeruginosa, the ethyl acetate fraction showed stronger antibacterial activity than other fractions and the extract. Moreover, the ethyl acetate fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell. However, in NO scavenging ability, the chloroform fraction was higher than the other fractions and the extract. In the lactate dehydrogenase (LDH) assay against RAW 264.7 cell, the extract and fractions were exhibited normal LDH release level as nontoxic result without the ehtyl acetate fraction of 100 ${\mu}g/ml$. As a result, the ethyl acetate fraction and chloroform fraction of the methanol extract from Paeoniae Radix could be applicable to functional materials for antibacterial and anti-inflammatory related fields, respectively.

Solvent effect on enzymatic steroid transformation

  • Kim, Doo-Ha;Lee, S.B.;Ryu, D.Y.
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.1-6
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    • 1980
  • As part of our endeavor to increase the productivity of steroid by enzymatic transformation of corticosteroids, attempts have been made to increase the solubility of steroids by using some organic solvents. When the solubility of steroids is the rate limiting factor in the steroid transformation, it was found that the use of solvents significantly improved the yield. Hydrocorisone as a substrate and 3-ketosteroid .DELTA.$^{1}$ dehydrogense as an immobilized whole cell enzyme were employed as the model system for this study. It was found that the yield of product, prodnisolone, goes through a maximum with an increase in the solvent concentration. At a high solvent concentration, the solvent showed a toxic effect and it causes a decrease in the product yield by the second order inhibition mechanism. Among the solvents evaluated, methanol and ethanol were found to be the best. These alcohols are not only good solvents but also showed minimal toxic effect. Based on the experimental results, it was concluded that the productivity of steroid can be increased by usign well selcted solvents systems for the enzymatic transformation of steroids.

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Identification and Characterization of Protein Encoded by orf382 as $\small{L}$-Threonine Dehydrogenase

  • Ma, Fei;Wang, Tianwen;Ma, Xingyuan;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.748-755
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    • 2014
  • In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).

Antioxidant and Neuronal Cell Protective Effects of Methanol Extract from Schizandra chinensis using an in vitro System (In vitro system에서 오미자 메탄올 추출물의 항산화 및 신경세포 보호효과)

  • Kim, Ji-Hye;Jeong, Chang-Ho;Choi, Gwi-Nam;Kwak, Ji-Hyun;Choi, Sung-Gil;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.712-716
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    • 2009
  • In this study, the antioxidant and neuronal cell protective effects of methanol extract from Schizandra chinensis were evaluated. The proximate composition and total phenolics content of the extract were as follows: 64.88% nitrogen free extract, 10.56% crude fiber, 10.22% moisture, 8.33% crude protein, 5.05% ash, 0.96% crude fat, and 83.04 mg/g of total phenolics. In assays the methanol extract of Schizandra chinensis presented ferric reducing/antioxidant power (FRAP) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity in a dose-dependent manner. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), the methanol extract showed protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into medium was also inhibited by various concentrations of extracts (68-80%). Cell viability after treatment of the methanol extract was higher than that shown for vitamin C ($100\;{\mu}M$) using a neutral red uptake (NRU) assay. Therefore, these data suggest that the methanol extract of Schizandra chinensis may be useful for neurodegenerative diseases including Alzheimer's disease.

Inhibitory Effects of Allium sacculiferum Max. Methanol Extracts on ROS Production and Lipid Accumulation during Differentiation of 3T3-L1 Cells (참산부추(Allium sacculiferum Max.) 메탄올 추출물의 지방세포 내 ROS 생성 및 지질 축적 억제 효능)

  • Choi, Hye-Young;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.6
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    • pp.822-828
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    • 2014
  • Allium sacculiferum Max. (ASM) is a perennial plant of the Liliaceae family and grows over the entire regions of Korea. Obesity is a serious health problem worldwide and has currently become a prevalent chronic disease. Adipocytes produced by preadipocyte differentiation during adipogenesis and adipocytes combined with abnormal accumulation cause obesity. Recently, intracellular reactive oxygen species (ROS) were shown to accelerate lipid accumulation in 3T3-L1 cells. In this study, we investigated the effects of ASM methanol extracts on ROS production and lipid accumulation in 3T3-L1 adipocytes. Our results indicate that the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of ASM methanol extracts increased in a dose-dependent manner. ASM methanol extracts suppressed ROS production and lipid accumulation during adipogenesis. In addition, ASM methanol extracts inhibited the mRNA expression of both pro-oxidant enzymes such as glucose-6-phosphate dehydrogenase as well as the transcription factors, including sterol regulatory element-binding proteins 1c, peroxisome proliferator-activated receptor ${\gamma}$, and CCAAT/enhancer-binding protein ${\alpha}$. Our results suggest that ASM methanol extracts inhibit ROS production and lipid accumulation by controlling ROS regulatory genes and adipogenic transcription factors. Thus, ASM has potent natural antioxidant, anti-adipogenic properties and have potential in the development of a potent anti-obesity agent.

Neuronal Cell Protective Effects of Methanol Extract from Cheonggukjang Using in vitro System (In vitro system에서 청국장 메탄올 추출물의 신경세포 보호효과)

  • Jeong, Chang-Ho;Kwak, Ji-Hyun;Kim, Ji-Hye;Choi, Gwi-Nam;Jeong, Hee-Rok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.6
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    • pp.768-772
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    • 2010
  • In this study, the neuronal cell protective effects of methanol extract from cheonggukjang were evaluated. The proximate composition and total phenolics of the methanol extract were 40.95% crude protein, 22.49% crude fat, 15.99% nitrogen free extract, 7.91% moisture, 6.74% crude ash, 5.92% crude fiber, and 28.43 mg/g of total phenolics. Intracellular ROS accumulation resulting from $H_2O_2$ treatment of PC12 cells was significantly reduced when methanol extract was present in the media compared to PC12 cells treated with $H_2O_2$ only. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT), the methanol extract showed protective effects against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into the medium was also inhibited. Furthermore, the inhibitory effect of the methanol extract against acetylcholinesterase was dose-dependent.

Effects of Dietary Prosomillet on cholesterol and Fatty Acid Metabolism in Rats Fed High Cholesterol Diets

  • Cho, Sung-Hee;Jung, Seung-Eun;Lee, Hye-Kyung;Ha, Tae-Youl
    • Nutritional Sciences
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    • v.3 no.1
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    • pp.25-30
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    • 2000
  • To study the effect of prosomillet (Panicum milaceum) on lipid metabolism, male Sprague-Dawley rats weighing 190$\pm$8g were fed six experimental diets for four weeks. The six diets based on AIN-76 composition consisted of one cholesterol-free(normal) and five 1%(w/w) cholesterol diets, i.e. control, two diets containing additional 0.3 and 0.6%(w/w) methanol extracts of prosomillet and another two diets containing 15 and 30% (w/w) prosomillet powder. There was no difference in weight gains between the groups but relative liver weights increased under the cholestrol diets. Plasma levels of total cholesterol and triglyceride(TG) decreased by 23-27% and by 37-52%, respectively, in the four prosomillet diet groups compared to those of the normal and control groups. Whereas in the liver, only TG levels decreased in the prosomillet diet groups. Fecal excretions of bile acid and cholesterol significantly with methanol extracts of prosomillet. There was a significant increase in the activity of hepatic microsomal cholesterol 7$\alpha$-hydroxylase when feeding 1% cholesterol but prosomillet in the diet, either as in the form of powder or methanol extract, appeared to have only slight additional effects, namely increases in enzyme activity. The activity of liver cytosolic glucose-6-phophate dehydrogenase (G6PDH) tended to be reduced with high cholesterol diets and dropped markedly by 15% using additional prosomillet powder. Those of the liver cytoxolic malic enzyme had a similar tendency to those of G6PDH. The results indicate that certain active components in prosmillet other than fiber have the potential to exert hypolipidemic effects via regulating cholesterol excretions and lipogenesis.

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Neuroprotective Effect of Extracts from Root Bark of Morus alba on Glutamate-induced Cytotoxicity in Neuronal Cells. (Glutamate가 유도하는 세포독성으로부터 신경세포를 보호하는 상백피 추출물의 효과)

  • Kim, Hyun-Jung;Kim, Ji-Hyun;Son, Eun-Soon;Lee, Jeung-Min;Park, Hae-Ryong
    • Journal of Life Science
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    • v.19 no.7
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    • pp.963-967
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    • 2009
  • This study evaluated the neuroprotective effect of extracts from the root bark of Morus alba (MA) against glutamate-induced cytotoxicity in neuronal cells. Glutamate-induced cytotoxicity was shown by MTT reduction assay. The neuroprotective effects of methanol, ethanol, and acetone extracts from MA against glutamate-induced cytotoxicity were measured. Among the three extracts, the methanolic extracts showed the highest protective effect, as determined by the results of an morphological assay, a lactate dehydrogenase release assay. Furthermore, the methanol extracts were fractionated sequentially with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited a neuroprotective effect against glutamate-stressed N18-RE-105 cells. Therefore, these results suggest that extracts of MA could be a new potential candidate as a protective substance against glutamate-induced cytotoxicity.

Anti-oxidative Effects of Dendrobii Herba on Toxic Agent Induced Kidney Cell Injury (석곡(石斛)의 항산화 효과)

  • Kim, Young-Gyun;Yang, Gi-Ho;Cho, Su-In
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.53-60
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    • 2005
  • Objectives : This study was carried out to determine if Dendrobii Herba have protective effect against cell injury induced by various toxic agents in rat kidney slices. Water(DWe) and methanol(DMe) extracts were prepared for this experiment. Methods : Cell injury was estimated by measuring lactate dehydrogenase(LDH). Lipid peroxidation was examined by measuring malondialdehyde, a product of lipid peroxidation. Results : DMe prevented the LDH release by $CCl_4$, menadione, tert-butyl hydroperoxide and mercury treatment in vitro in kidney slices, but DWe prevented the LDH release by $CCl_4$ and mercury. DMe also prevented reduction in GSH and lipid peroxidation induced by $CCl_4$ and mercury. Conclusion : Thus, DMe may have more powerful efficacy on anti-oxidative effects when compared with DWe. And further studies have to be followed concerned with extraction of Dendrobii Herba and its change of effects.

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