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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Applied Biological Chemistry
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Journal DOI :
The Korean Society for Applied Biological Chemisty
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Volume 45, Issue 4 - 00 2002
Volume 45, Issue 3 - 00 2002
Volume 45, Issue 2 - 00 2002
Volume 45, Issue 1 - 00 2002
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Sequence Analysis of a Putative Membrane Transporter cDNA of Arabidopsis thaliana
Park, Sang-Gyu ; Thornburg, Robert W. ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 153~159
Six different clones of Arabidopsis thaliana expressed sequence tags (ESTs) were sequenced, among which ATTS1488 clone appeared to code a putative membrane transporter of purines. Comparison of the nucleotide sequence of ATTS1488 with that of a genomic putative membrane transporter gene showed that one nucleotide (A) was missing in the EST clone and another nucleotide (T) of the genomic sequences was missing in the ATTS1488 sequences. These changes caused three amino acid differences between genomic DNA and cDNA. Deduced amino acid sequence of the cDNA has a 99.4% identity with that of the membrane transporter genomic DNA, and a 51% amino acid identity with those of rice (Oryza sativa) permease 1 and another putative membrane transporter of Arabidopsis. Prediction of membrane protein structure indicates that the EST clone has 13 trans-membrane domains in the deduced amino acid sequences.
Extracellular Production of 5-Aminolevulinic Acid and Affinity Purification of Bradyrhizobium japonicum 5-Aminolevulinic Acid Synthase in Escherichia coli
Ha, Suk-Bong ; Lee, Da-Eun ; Back, Kyung-Whan ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 160~163
5-Aminolevulinic acid synthase (ALAS) catalyzes the condensation of glycine and succinyl-CoA into 5-aminolevulinic acid (ALA) together with the release of CO
and CoA. An Escherichia coli expression system was used to generate hexahistidyl-tagged Bradyrhizobium japonicum ALAS, which was readily purified using single step affinity chromatography. The gene encoding ALAS was cloned in frame with the N-terminal hexahistidine tag in cloning vector pET-28b(+). ALAS accumulated up to 70-80% of the total soluble protein when the expression construct was induced in E. coli, and was purified to over 98% homogeneity. Recombinant ALAS was active in E. coli producing up to 2 mM extracellular ALA 3 h after the induction with IPTG. Purified recombinant ALAS was used as an antigen to produce a mouse antibody that crossreacts specifically with ALAS.
Molecular Cloning and Functional Expression of Acyl-Acyl Carrier Protein Thioesterase cDNA from Iris germanica
Kim, Kyung-Hwan ; Kim, Jung-Bong ; Lim, Myung-Ho ; Lee, Jai-Youl ; Hwang, Young-Soo ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 164~169
The plant acyl-acyl carrier protein (ACP) thioesterases play an essential role in chain termination during de novo fatty acid synthesis. An acyl-ACP thioesterase cDNA, designated IgFatB1, from Iris germanica, which has an unusual fatty-acyl composition containing 57% myristic and 15.5% lauric acids in root, was isolated from Iris cDNA root library. IgFatB1, a cDNA coding 421 amino acids, showed high degree of sequence similarity to corresponding enzymes from various plants, and expressed in E. coli, functional recombinant protein has thioesterase (TE) activity for 16:0-ACP substrate. When expressed in beta-oxidation mutants of E. coli, lipid analysis revealed that cells transformed with the IgFatB1 fusion plasmid produced high amount of free fatty acids, mostly consisting of 14:0 and some 16:
Apigenin, Tyrosinase Inhibitor Isolated from the Flowers of Hemisteptia lyrata Bunge
Ha, Tae-Joung ; Hwang, Sun-Woo ; Jung, Hwa-Jin ; Park, Ki-Hun ; Yang, Min-Suk ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 170~172
From the flowers of Hemisteptia lyrata Bunge, two flavonoids, apigenin (1) and acacetin (2), were isolated. Their structures were established by UV, one- and two-dimensional NMR, and mass spectroscopic methods. Apigenin showed significant inhibition against mushroom tyrosinase with an IC
value of 4.56
, and appeared to inhibit the polyphenol oxidase activity of tyrosinase in a competitive manner (K
M) when L-tyrosine was used as a substrate, but did not inhibit the melanin production of Streptomyces bikiniensis.
Effects of Extracts from Rhus Verniciflua S. on Activity of Catalase and TPA Mediated-Apoptosis in Cultured Mouse Hepatocytes
Lim, Kye-Taek ; Heo, Kyung-Sun ; Shim, Jae-Han ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 173~179
The ethanol extract of Rhus verniciflua S. was subsequently isolated and fractioned into two portions using H
O and 99% ethanol as elution buffers through silica gel column chromatography. To study the antioxidative effect of Rhus verniciflua S. extracts, cultured hepatocytes were exposed to hydroxyl radical generated by 20 mU ml
glucose oxidase for 4 h in the presence or absence of water or ethanol eluted extracts. Addition of 100
water extract to the culture medium protected from hydroxyl radical-mediated cytotoxicity of hepatocytes almost equivalently to the control. When the hepatocytes after culture for 24 h were incubated with 100
water or ethanol extract without glucose oxidase for 4 h, activity of catalase was increased by 1.40- and 1.31-fold, compared to the control, whereas with the commercial catalase (100 units ml
) was 1.64-fold. These results show that in vitro both extracts has a potential reducing cytotoxicity effect on hydroxyl radical and enhancing activities of the cellassociated catalase in cultured hepatocytes. Apoptosis induced by 12-O-tetradecanoylphorbol 13-acetate (TPA, 100 nM) in mouse hepatocytes were blocked by the addition of 100
of water or ethanol extract. In the biological effects of two extracts, water extract was more effective than that of ethanol extract.
A Protocol for Batch Fermentation for Production of Soluble Alpha Interferon by Recombinant Escherichia coli
Kim, Dong-Chung ; Jung, Jin ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 180~182
Improvement of alpha interferon (IFN
) production was attempted using recombinant E. coli by optimizing fermentation conditions. A protocol that ensures an increased productivity of IFNa mostly in active, soluble form without affecting the growth activity of cells was devised. Although the cells grew well in a neutral medium, a slight decrease in medium pH, i.e. to pH 6.7, resulted in a significant increase in the production of soluble IFN
. The growth activity and total IFN
productivity were not altered by a change in fermentation temperature in the range of 30-37
. Ratio of soluble form to inclusion body increased with decreasing temperature, with over 90% of total interferon being of soluble form when cultivated at 30
. An increase in the nitrogen concentration of Terrific Broth-based medium supplemented with L-Ieucine gave remarkably improved results. A fermentation protocol derived from these observations enabled us to enhance the productivity level of IFN
IU/-broth by 14 h fermentation.
Hepatocyte Primary Cell Culture as a Model System for Investigating the Effect of Stearic Acid on Cholesterol and Glycerol Lipid Metabolism
Pai, Tong-Kun ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 183~186
The optimum condition of rat hepatocyte primary cell culture for use as a model system to study the role of various fatty acids on cholesterol and glycerol lipid metabolism was determined. Up to 1.5
cells/well, a linear relationship (r = 0.96) was observed between the cell number and the quantity of cellular protein recovered after the incubation. Stearic acid bound to bovine serum albumin (BSA) at high levels in the medium (>97%) at all fatty acid-BSA ratios. Results reveal that the cellular protein (mg) can be used as the base unit to express the fatty acid metabolism in the rat hepatocyte primary culture and that stearic acid-BSA ratio of 5.6 : 1 is optimum for studying the effects of various fatty acids on lipid metabolism when considering the difficulty of handling the medium at high ratios.
Detection of Irradiated Medicinal Herbs by Viscosity Measurement and ESR Spectroscopy
Yi, Jin-Hee ; Yang, Jae-Seung ; Song, Kyung-Bin ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 187~190
Ionizing radiation can be used to prevent microbial spoilage of herb and extend its shelf life. To elucidate the physicochemical change of herb caused by irradiation. electron spin resonance (ESR) spectroscopy and viscosity measurement were performed after irradiation of two typical medicinal herbs, Rehmannia giutinosa and Havenia dulcis Thumb. With increasing irradiation dose up to 10 kGy, viscosity values of all the samples decreased proportionally. ESR spectral signal intensity increased with increasing irradiation dose. These results clearly imply that ESR analysis and viscosity measurement can be used for a simple and reliable detection of irradiated herbs.
Water Stress Enhances Nitrate Accumulation in Lettuce at Low Light Intensity
Jin, Sun-Jae ; Cho, Hyun-Jong ; Chung, Jong-Bae ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 191~195
Accumulation of nitrate in edible crops is undesirable due to potential risks to human health. Since nitrate has a role in the osmotic regulation of plants, salt accumulation in soil is expected to stimulate nitrate accumulation in plants. Lettuce was grown in nutrient solutions of different water potentials prepared by adding PEG in a greenhouse, and the effect of water stress on nitrate accumulation in lettuce was investigated. Content of nitrate in the lettuce increased significantly as water potential in the nutrient solution decreased under low light intensity and ample supply of nitrate in root media. Total soluble sugar and oxalate contents also increased with water stress treatments. These results indicate that higher nitrate content in plants under water stress is a positive accumulation to adapt to the water stress condition. Phosphate, Cl, and
contents in lettuce were not significantly different among the nutrient solutions of different water potentials. Among the cations, K content was significantly increased in plants under water stress and Na, Ca, and Mg contents were not influenced. The accumulation of nitrate in vegetables grown in plastic film houses is known to be due to the heavy fertilization and low light intensity, but salt accumulation in the soil, which can lower soil water potential, is expected to stimulate the nitrate accumulation further.
Effects of Organic Acids on the Growth of a Phytopathogenic Fungus, Colletotrichum Species
Kang, Han-Chul ; Go, Seung-Joo ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 196~201
Acetic, oxalic, malic, and citric acid anions significantly inhibited the growth of Colletotrichum, a phytopathogenic fungus in a concentration- dependent manner at pH 5.0. Acetic acid showed the strongest inhibition with no growth at 50 mM. The acetic or citric acid-mediated growth inhibition was further confirmed with three additional strains, C. cocodes, C. gloeosporioides, and C. dematium. Optimum growth of Colletotrichum was observed around pH 6.0; in the presence of acetic or citric acid, the growth inhibition accelerated along with a decrease in pH from 6.0 to 4.0, suggesting that the inhibition might be more enhanced by undissociated form of organic acids. Colletotrichum did not grow with 50 mM acetic acid at pH 3.0. After elimination of acetic acid, the fungus began to grow in a normal culture medium. Addition of acetic acid into the culture medium stopped the fungus from utilizing glucose, suggesting that the growth inhibition may be attributed to a metabolic inhibition, and not to the cell damage. The addition of acetic and ethylenediaminetetraacetic acids (EDTA) showed a synergistic effect in the fungal growth inhibition. Controlling of Colletotrichum could be developed using acetic acid, which is generally less dangerous than chemical reagents.
Fungicidal Activities of 67 Herb-Derived Oils against Six Phytopathogenic Fungi
Cho, Jang-Hee ; Kim, Jin-Cheol ; Kim, Moo-Key ; Lee, Hoi-Seon ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 202~207
The oils from 67 herbs were tested for their fungicidal activities against 6 phytopathogenic fungi in a greenhouse at 10 and 5 mg/pot. The efficacy varied with both the plant pathogen and herb oil species used. The oils of Anthemis nobilis and Rosemarinus officinalis gave over 80% control value (CV) against Pyricularia grisea, and that of Curcuma longa exhibited potent fungicidal activity (CV >90%) against Rhizoctonia solani. In a test with Botrytis cinerea, the oils of Cuminum cyminum and Lavendula augustifolia exhibited strong fungicidal activities. Against Phytophthora infestans, the oil of C. longa showed strong fungicidal activity. The oils of Dioscorea villosa and Vetiveria zizanioides produced strong fungicidal activities against Puccinia recondita, and those of C. longa and Melaleuca viridiflora showed strong activities against Erysiphe graminis. Potent fungicidal activities were produced from the oil of C. cyminum against SSR and SRR, and L. augustifolia against SSR and RSR strains. As a naturally occurring fungicide, these herb-derived oils could be useful as new fungicidal products against phytopathogenic fungi.
Screening of Prolyl Endopeptidase Inhibitors from Natural Products
Lee, Si-Young ; Lee, Jung-Han ; Hahn, Tae-Ryong ; Paik, Young-Sook ;
Journal of Applied Biological Chemistry, volume 45, issue 4, 2002, Pages 208~210
Prolyl endopeptidase (PEP, EC 18.104.22.168) plays an important role in the degradation of biologically active proline-containing neuropeptides such as vasopressin, substance P, and thyrotropin-releasing hormone (TRH) and has been speculated to participate in learning and memory processes. In the course of search for anti-amnestic constituents, 105 fractions were prepared from 20 traditional medicinal plants and screened for PEP inhibitors. Specified fractions of Eugenia caryophyllata (flos), Ganoderma lucidum (fungus), Ginkgo biloba (folium), Paeonia suffruticosa (cortex), Pueraria thunbergiana (flos), and Rhodiola sachalinensis (radix) showed 54-99% PEP inhibitory activities at 4