(Vol.69 No.02 2005)
The Administration of Retinoic Acid Down-Regulates cAMP-Responsive Element Modulator (CREM) mRNA in Vitamin A-Deficient Testes
Mami Matsuda1, Tomomasa Hyoudou2, Masanori Kadowaki1, Kaori Onuki1, Shoichi Masushige1,2, and Satoshi Kida1,2 p.261
Effects of Fermentation Temperature on the Content and Composition of Isoflavones and 2-Glucosidase Activity in Sufu
Li-jun Yin1,2, Li-te Li1, Huan Liu1, Masayoshi Saito2, and Eizo Tatsumi2 p.267
The Necessity of Niacin in Rats Fed on a High Protein Diet
Naoko Kimura, Tsutomu Fukuwatari, Ryuzo Sasaki, and Katsumi Shibata p.273
Structural and Functional Characterization of Polyphenols Isolated from Acerola (Malpighia emarginata DC.) Fruit
Takayuki Hanamura1, Toshihiko Hagiwara1, and Hirokazu Kawagishi2 p.280
Three Oxygenated Cyclohexenone Derivatives Produced by an Endophytic Fungus
Yoshihito Shiono1, Tetsuya Murayama1, Koetsu Takahashi2, Katsuhide Okada3, Shigeyoshi Katohda1, and Michimasa Ikeda1 p.287
Indole Hydroxylation by Bacterial Cytochrome P450 BM-3 and Modulation of Activity by Cumene Hydroperoxide
Qing-Shan Li1, Jun Ogawa1, Rolf D. Schmid2, and Sakayu Shimizu1 p.293
Short-Term Feeding of Fish Oil Down-Regulates the Expression of Pyruvate Dehydrogenase E1 Alpha Subunit mRNA in Mouse Brain
Yoshihito Nogusa1, Noriyuki Yanaka1, Naoki Sumiyoshi1, Yurika Kaseda1, Nobuyo Tsuboyama-kasaoka2, Osamu Ezaki2, and Norihisa Kato1 p.301
Nobiletin, a Citrus Flavonoid, Down-Regulates Matrix Metalloproteinase-7 (matrilysin) Expression in HT-29 Human Colorectal Cancer Cells
Kyuichi Kawabata, Akira Murakami, and Hajime Ohigashi p.307
Biosynthetic Origin of the Carbon Skeleton and Nitrogen Atom of Pamamycin-607, a Nitrogen-Containing Polyketide
Makoto Hashimoto, Haruhiko Komatsu, Ikuko Kozone, Hiroshi Kawaide, Hiroshi Abe, and Masahiro Natsume p.315
Formation of trans-Verbenol and Verbenone from ±-Pinene Catalysed
by Immobilised Picea abies Cells
Tomäa Vank1, Jan Hal\ik1, Radmila VaHkovä2, and Irena Valterovä1 p.321
Control of Tomato Bacterial Wilt without Disinfection Using a New Functional Polymer That Captures Microbial Cells Alive on the Surface and Is Highly Biodegradable
Nariyoshi Kawabata1, Hitoshi Kishimoto2, Takayuki Abe1, Tomokatsu Ikawa2, Katsumi Yamanaka2, Hisaya Ikeuchi1, and Chizuko Kakimoto1 p.326
Distribution of Gibberellins and Expressional Analysis of GA 20-oxidase Genes of Morning Glory during Fruit Maturation
Akira Nakayama, Masatoshi Nakajima, and Isomaro Yamaguchi p.334
On a Salmon (Onchorhynchus keta) Liver RNase, Belonging to RNase T2 Family: Primary Structure and Some Properties
Rie Suzuki1, Sachiko Kanno1, Yuko Ogawa1, Masanori Iwama3, Tsutomu Tsuji2, Kazuko Ohgi1, and Masachika Irie1 p.343
Vaticanol C-Induced Cell Death Is Associated with Inhibition of Pro-Survival Signaling in HL60 Human Leukemia Cell Line
Kenji Ohguchi1, Yukihiro Akao1, Kenji Matsumoto1, Toshiyuki Tanaka2, Tetsuro Ito2, Munekazu Iinuma3, and Yoshinori Nozawa1 p.353
2-Hydroxyergothioneine, a New Ergothioneine Derivative from the Mushroom Lyophyllum connatum, and Its Protective Activity against Carbon Tetrachloride-Induced Injury in Primary Culture Hepatocytes
Chieko Kimura1, Manabu Nukina1,2, Kiharu Igarashi1,2, and Yoko Sugawara3 p.357
Gene Cloning, Overproduction, and Characterization of Thermolabile Alkaline Phosphatase from a Psychrotrophic Bacterium
Yutaka Suzuki1, Yoichi Mizutani1, Tadao Tsuji1, Naoto Ohtani1, Kazufumi Takano1,2, Mitsuru Haruki1, Masaaki Morikawa1, and Shigenori Kanaya1 p.364
Enantioselective Total Synthesis of Enokipodins AD, Antimicrobial Sesquiterpenes Produced by the Mushroom, Flammulina velutipes
Mana Saito and Shigefumi Kuwahara p.374
Molecular Dissection of the Promoter of the Light-Induced and Circadian-Controlled APRR9 Gene Encoding a Clock-Associated Component of Arabidopsis thaliana
Shogo Ito, Norihito Nakamichi, Akinori Matsushika, Toru Fujimori, Takafumi Yamashino, and Takeshi Mizuno p.382
Synthesis and Algicidal Activity of (+)-Cyanobacterin and Its Stereoisomer
Fumito Ishibashi1, Suhwan Park1, Takako Kusano1, and Kazuyoshi Kuwano2 p.391
Microstructures of Bread Dough and the Effects of Shortening on Frozen Dough
Shigeo Aibara1, Noriko Ogawa2, and Masaaki Hirose1 p.397
Note
Sericin Enhances Attachment of Cultured Human Skin Fibroblasts
Kozo Tsubouchi, Yumiko Igarashi, Yoko Takasu, and Hiromi Yamada p.403
Note
Cloning, Expression in Yeast, and Functional Characterization of CYP76A4, a Novel Cytochrome P450 of Petunia That Catalyzes ( É -1)-Hydroxylation of Lauric Acid
Katsutomo Tamaki1, Hiromasa Imaishi2, Hideo Ohkawa3, Kiyoharu Oono2, and Mikihiro Sugimoto1 p.406
Note
Circadian-Associated Rice Pseudo Response Regulators (OsPRRs): Insight into the Control of Flowering Time
Masaya Murakami1, Akinori Matsushika1, Motoyuki Ashikari2, Takafumi Yamashino1, and Takeshi Mizuno1 p.410
Note
Transient RNAi Induction against Endogenous Genes in Arabidopsis Protoplasts Using in Vitro-Prepared Double-Stranded RNA
Chung-Il An, Aki Sawada, Yoshiaki Kawaguchi, Ei-ichiro Fukusaki, and Akio Kobayashi p.415
Note
Cold-Active Polygalacturonase from Psychrophilic-Basidiomycetous Yeast Cystofilobasidium capitatum Strain PPY-1
Tomoyuki Nakagawa, Toshinori Nagaoka, Tatsuro Miyaji, and Noboru Tomizuka p.419
Note
Structural and Functional Analysis of pTB6 from Bifidobacterium longum
Kunihito Tanaka, Kazuya Samura, and Yasunobu Kano p.422
Note
PRR5 (PSEUDO-RESPONSE REGULATOR 5) Plays Antagonistic Roles to CCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) in Arabidopsis thaliana
Toru Fujimori, Eriko Sato, Takafumi Yamashino, and Takeshi Mizuno p.426
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The Administration of Retinoic Acid Down-Regulates cAMP-Responsive Element Modulator
(CREM) mRNA in Vitamin A-Deficient Testes
Mami Matsuda1, Tomomasa Hyoudou2, Masanori Kadowaki1, Kaori Onuki1, Shoichi Masushige1,2, and Satoshi Kida1,2
1Department of Agricultural Chemistry, Faculty of Agriculture, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan
2Department of Bioscience, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, JapanStudies using genetics and vitamin A deficiency (VAD) have shown that vitamin A and retinoids play essential roles in spermatogenesis at the pre-meiotic stage. To understand the mechanisms of control in spermatogenesis by retinoic acid, we investigated whether retinoic acids regulate the expression of downstream transcription factors that are essential for spermatogenesis. In this study, we found that administration of all-trans retinoic acid (ATRA) or retinol to VAD rats down-regulates the testicular mRNA levels of the cAMP responsive element modulator (CREM), an essential transcription factor for spermatogenesis. Conversely, depletion of retinoids from the diet leds to an up-regulation of CREM expression in adult testes. In addition, RT-PCR analysis indicated that ATRA specifically represses the expression of the activator spliced variant of CREM (CREMÄ ). These results suggest that retinoids function as a negative regulator of CREM expression in testes.
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Effects of Fermentation Temperature on the Content and Composition of Isoflavones and 2-Glucosidase Activity in Sufu
Li-jun Yin1,2, Li-te Li1, Huan Liu1, Masayoshi Saito2, and Eizo Tatsumi2
1College of Food Science and Nutritional Engineering, Box 40, China Agricultural University, Qinghua East Road, Beijing 100083, China
2Food Science and Technology Division, Japan International Research Center for Agricultural Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8686, JapanSufu is a popular fermented tofu product in China. The low quality of sufu produced in the hot summer is a big problem in sufu manufacture, so we prepared sufu at two different temperatures, 26 ℃ as normal and 32 ℃ as high temperature, and the effects of temperature on isoflavones and 2-glucosidase activity were investigated. Fermentation temperature did not cause significant differences in the recovery of isoflavones, but resulted in a different redistribution of isoflavone isomers in sufu. Sufu fermented at 26 ℃ was richer in isoflavone aglycones than at 32 ℃; the enrichment of isoflavone aglycones might have the advantage of enhancing the physiological function. No 6 -O-malonyl-glucosides were detected in sufu fermented at 26 ℃, whereas some 6 -O-malonyl-glucosides were found at 32 ℃. A fermentation temperature of 26 ℃ benefited the 2-glucosidase production by fungi, which contributed to valid conversion from 2-glucosides to aglycones. It was also found that 2-glucosidase converted 2-glucosides more effectively than 6 -O-malonyl-glucosides and 6 -O-acetyl-glucosides into aglycones.
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The Necessity of Niacin in Rats Fed on a High Protein Diet
Naoko Kimura, Tsutomu Fukuwatari, Ryuzo Sasaki, and Katsumi Shibata
Laboratories of Food Science and Nutrition, Department of Life Style Studies, School of Human Cultures, The University of Shiga Prefecture, 2500 Hassakacho, Hikone, Shiga 522-8533, Japan
It is known that niacin itself is not necessary in rats when tryptophan is given in adequate amounts, because rats can biosynthesize niacin from tryptophan. In our experiment, young rats were fed on a 20%, 40%, 60%, or 70% casein diet with or without niacin. The rats fed on the 20%, 40%, and 60% casein diets did not require niacin for growth, but the rats fed on the 70% casein diet needed it. This phenomenon was attributed to the supposition that liver aminocarboxymuconatesemialdehyde decarboxylase activities increased according with the dietary casein levels. The conversion ratio of tryptophanniacin in rats fed on the 70% casein diet became extremely low, and then the rats needed niacin.
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Structural and Functional Characterization of Polyphenols Isolated from Acerola (Malpighia emarginata DC.) Fruit
Takayuki Hanamura1, Toshihiko Hagiwara1, and Hirokazu Kawagishi2
1Research and Development Division, Processed Foods Company, Nichirei Corporation, 9 Shinminato, Mihama-ku, Chiba-shi, Chiba 261-8545, Japan
2Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, JapanTwo anthocyanins, cyanidin-3-±-O-rhamnoside (C3R) and pelargonidin-3-±-O-rhamnoside (P3R), and quercitrin (quercetin-3-±-O-rhamnoside), were isolated from acerola (Malpighia emarginata DC.) fruit. These polyphenols were evaluated based on the functional properties associated with diabetes mellitus or its complications, that is, on the radical scavenging activity and the inhibitory effect on both ±-glucosidase and advanced glycation end product (AGE) formation. C3R and quercitrin revealed strong radical scavenging activity. While the inhibitory profiles of isolated polyphenols except quercitrin towards ±-glucosidase activity were low, all polyphenols strongly inhibited AGE formation.
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Three Oxygenated Cyclohexenone Derivatives Produced by an Endophytic Fungus
Yoshihito Shiono1, Tetsuya Murayama1, Koetsu Takahashi2, Katsuhide Okada3, Shigeyoshi Katohda1, and Michimasa Ikeda1
1Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, Tsuruoka-shi, Yamagata 997-8555, Japan
2Department of Bioenvironment, Faculty of Agriculture, Yamagata University, Tsuruoka-shi, Yamagata 997-8555, Japan
3Laboratory of Food Science, Faculty of Education, Yamagata University, Yamagata-shi, Yamagata 990-8560, JapanThree cyclohexenone derivatives, (4S,5S,6S)-5,6-epoxy-4-hydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (1), (4R,5R)-4,5-dihydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (2), and (4R,5S,6R)-4,5,6-trihydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (3), were isolated from unpolished rice fermented with an xylariaceous endophytic fungus (strain YUA-026). The structures of three compounds were established on the basis of spectroscopic analyses and chemical conversion. The minimum inhibitory concentrations of 1 and 3 were 100 ¼g/ml and 400 ¼g/ml against Staphylococcus aureus, 100 ¼g/ml and 200 ¼g/ml against Pseudomonas aeruginosa, and 200 ¼g/ml and >400 ¼g/ml against Candida albicans, respectively. In addition, 1 and 3 exhibited phytotoxic activity against lettuce.
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Indole Hydroxylation by Bacterial Cytochrome P450 BM-3 and Modulation of Activity by Cumene Hydroperoxide
Qing-Shan Li1, Jun Ogawa1, Rolf D. Schmid2, and Sakayu Shimizu1
1Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
2Institut für Technische Biochemie, Universität Stuttgart, Allmandring 31, D-70459 Stuttgart, GermanyCytochrome P450 BM-3 from Bacillus megaterium catalyzed NADPH-supported indole hydroxylation under alkaline conditions with homotropic cooperativity toward indole. The activity was also found with the support of H2O2, tert-butyl hydroperoxide (tBuOOH), or cumene hydroperoxide (CuOOH). Enhanced activity and heterotropic cooperativity were observed in CuOOH-supported hydroxylation, and both the Hill coefficient and substrate concentration required for half-maximal activity in the CuOOH-supported reaction were much lower than those in the H2O2-, tBuOOH-, or NADPH-supported reactions. CuOOH greatly enhanced NADPH consumption and indole hydroxylation in the NADPH-supported reaction. However, when CuOOH was replaced by tBuOOH or H2O2, heterotropic cooperativity was not observed. Spectral studies also confirmed that CuOOH stimulated indole binding to P450 BM-3. Interestingly, a mutant enzyme with enhanced indole-hydroxylation activity, F87V (Phe87 was replaced by Val), lost homotropic cooperativity towards indole and heterotropic cooperativity towards CuOOH, indicating that the active-site structure affects the cooperativities.
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Short-Term Feeding of Fish Oil Down-Regulates the Expression of Pyruvate Dehydrogenase E1 Alpha Subunit mRNA in Mouse Brain
Yoshihito Nogusa1, Noriyuki Yanaka1, Naoki Sumiyoshi1, Yurika Kaseda1, Nobuyo Tsuboyama-kasaoka2, Osamu Ezaki2, and Norihisa Kato1
1Department of Bioresource Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima 739-8528, Japan
2Division of Clinical Nutrition, National Institute of Health and Nutrition, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8636, JapanPrevious studies have suggested that docosahexaenoic acid (DHA), contained in fish oil, prevents brain disease. In the current study, the effect of fish oil feeding on gene expression in the brain was investigated by suppression subtractive hybridization. We found that pyruvate dehydrogenase E1 alpha (PDHE1±) mRNA expression is down-regulated by fish oil feeding. We examined whether the expression of PDHE1± mRNA is altered by DHA treatment in differentiated PC12 cells. PDHE1± mRNA was reduced by supplementation of DHA with a significant decrease in cellular ATP level. These results indicate that fish oil feeding might modulate energy metabolism in the brain.
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Nobiletin, a Citrus Flavonoid, Down-Regulates Matrix Metalloproteinase-7 (matrilysin) Expression in HT-29 Human Colorectal Cancer Cells
Kyuichi Kawabata, Akira Murakami, and Hajime Ohigashi
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
Overexpression of matrix metalloproteinases (MMPs) is associated with cancer metastasis. We assessed mRNA expression of MMPs in six human colorectal cancer cell lines and found a considerable level of MMP-7 expression in HT-29 cells. Next, we searched for natural and synthetic compounds that cause a reduction in the production of proMMP-7 protein, and found that nobiletin (NOB), quercetin, valeryl salicylate, and sulindac sulfone demonstrated marked inhibition. Importantly, NOB attenuated proMMP-7 protein and its mRNA expression both concentration- and time-dependently via a reduction of activator protein-1 (AP-1) DNA binding activity, suggesting it as a promising agent for suppression of cancer cell invasion and metastasis through MMP-7 gene repression.
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Biosynthetic Origin of the Carbon Skeleton and Nitrogen Atom of Pamamycin-607, a Nitrogen-Containing Polyketide
Makoto Hashimoto, Haruhiko Komatsu, Ikuko Kozone, Hiroshi Kawaide, Hiroshi Abe, and Masahiro Natsume
Department of Plant Protection, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Saiwai-cho, Fuchu, Tokyo 183-8509, Japan
The biosynthesis of pamamycin-607 (PM-607), a sixteen-membered macrodiolide compound, was studied with 13C- and 15N-labeled precursor units in Streptomyces alboniger. Feeding experiments with 13C-labeled acetate or propionate indicate that the carbon skeleton of PM-607 was derived from six acetate, four propionate and three succinate units. MS analyses of 15N-labeled PM-607 suggest that the nitrogen atom in PM-607 was derived from the ±-amino group of an amino acid.
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Formation of trans-Verbenol and Verbenone from ±-Pinene Catalysed by Immobilised Picea abies Cells
Tomáa Vank1, Jan Hal\ik1, Radmila VaHková2, and Irena Valterová1
1Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague, Czech Republic
2Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 135, 165 02 Prague, Czech RepublicBoth enantiomers and the racemate of ±-pinene were transformed by Picea abies cells immobilised on alginate. The main products were cis- and trans-verbenol, the later being further transformed to verbenone. The enantiomeric purity of each product more or less corresponded to that of the substrate. Transformation by free cells was faster than that by the immobilised cells. The ratio of products differed to some extent between the transformation by free and immobilised cells.
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Control of Tomato Bacterial Wilt without Disinfection Using a New Functional Polymer That Captures Microbial Cells Alive on the Surface and Is Highly Biodegradable
Nariyoshi Kawabata1, Hitoshi Kishimoto2, Takayuki Abe1, Tomokatsu Ikawa2, Katsumi Yamanaka2, Hisaya Ikeuchi1, and Chizuko Kakimoto1
1Department of Materials Science, Faculty of Engineering, University of Shiga Prefecture, Hassaka-cho, Hikone, Shiga 522-8533, Japan
2Department of Chemistry, Faculty of Engineering and Design, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, JapanThis report describes a green chemical method for controlling soil-borne plant diseases without disinfection using an equimolar copolymer of N-benzyl-4-vinylpyridinium chloride with styrene (PBVP-co-ST) that captures microbial cells alive on the surface and is highly biodegradable. Tomato bacterial wilt caused by Ralstonia solanacearum was controlled by the addition of sawdust coated with PBVP-co-ST prior to transplantation. This effected 87% reduction in appearance and 89% reduction in the index of symptom under appropriate conditions. The coated sawdust did not exhibit bactericidal activity. The half-life of PBVP-co-ST was 5.6 d when treated with activated sludge in soil. The disease control was explained in terms of reduction of infectious contact between the roots of tomato and the cells of R. solanacearum due to coagulation-like interaction between microbial cells and the coated sawdust, in addition to capture of microbial cells by the coated sawdust.
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Distribution of Gibberellins and Expressional Analysis of GA 20-oxidase Genes of Morning Glory during Fruit Maturation
Akira Nakayama, Masatoshi Nakajima, and Isomaro Yamaguchi
Department of Applied Biological Chemistry, University of Tokyo, Bunkyo, Tokyo 113-8657, Japan
Gibberellins A1/3 (GA1/3) and GA20 appeared earlier in surrounding tissues (pericarps/carpel/placenta) than in developing seeds of morning glory. The content of GA1/3 became higher in seeds than in the surrounding tissues at 9 days after anthesis (DAA), while that of GA20 stayed lower in seeds even at 12 DAA, suggesting the possibility that GA20 was translocated into seeds from the surrounding tissues and converted to GA1/3. The site of biosynthesis of GA20 in the fruits was determined by RNA-blotting and in situ hybridization of GA 20-oxidase genes (InGA20ox1, InGA20ox2). InGA20ox1 was not expressed in the surrounding tissues but in seeds, while no signal due to InGA20ox2 was detected in neither tissue. The expression of InGA20ox1 started in the seed coat near the hilum and spread in the seed coat like those of GA 3-oxidase and GA-inducible ±-amylase genes. These observations suggest that GA biosynthesis is tissue-specifically and time-dependently regulated in the fruit of morning glory.
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On a Salmon (Onchorhynchus keta) Liver RNase, Belonging to RNase T2 Family: Primary Structure and Some Properties
Rie Suzuki1, Sachiko Kanno1, Yuko Ogawa1, Masanori Iwama3, Tsutomu Tsuji2, Kazuko Ohgi1, and Masachika Irie1
1Department of Applied Microbiology, Hoshi University, 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501, Japan
2Microbiology, Faculty of Pharmaceutical Sciences, Hoshi University, 2-4-41 Ebara, Shinagawa-ku, Tokyo 142-8501, Japan
3Nagaoka National College of Technology, 888 Nishikatagai, Nagaoka, Niigata-ken 950-8532, JapanA base-nonspecific and acid ribonuclease (RNase Ok2) was purified from the liver of a salmon (Oncorhnchus keta) to a homogeneous state by SDSPAGE. The primary structure of RNase Ok2 was determined by protein chemistry and molecular cloning. The RNase Ok2 was a glycoprotein and consisted of 216 amino acid residues. Its molecular mass of protein moiety was 25,198, and its amino acid sequence showed that it belongs to the RNase T2 family of enzymes. The optimal pH of RNase Ok2 was around 5.5. The base preferences at the B1 and B2 sites were estimated from the rates of hydrolysis of 16 dinucleoside phosphates to be G>>A>U, C, and G>A>U>C respectively.
In this enzyme, one of the three histidine residues which have been thought to be important for catalysis of RNase Rh, a typical RNase of this family of enzymes, His104 was replaced by tyrosine residue. Based on the results, the role of H104, which has been proposed to be a phosphate binding site with a substrate, was reconsidered, and we proposed a revised role of this His residue in the hydrolysis mechanism of RNase T2 family enzymes.
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Vaticanol C-Induced Cell Death Is Associated with Inhibition of Pro-Survival Signaling in HL60 Human Leukemia Cell Line
Kenji Ohguchi1, Yukihiro Akao1, Kenji Matsumoto1, Toshiyuki Tanaka2, Tetsuro Ito2, Munekazu Iinuma3, and Yoshinori Nozawa1
1Gifu International Institute of Biotechnology, 1-1 Naka-Fudogaoka, Kakamigahara, Gifu 504-0838, Japan
2Gifu Prefectural Institute of Health and Environmental Sciences, Kakamigahara, Gifu 504-0838, Japan
3Gifu Pharmaceutical University, Mitahora-higashi, Gifu 502-5858, JapanRecently, we found that vaticanol C (a resveratrol tetramer), which was isolated from stem bark of Dipterocarpaceae, exhibited growth suppression and induction of apoptosis via the loss of mitochondrial membrane potential and consequent caspases activation. The detailed mechanisms are not clearly understood. We decided to attempt to gain further insight into the mechanisms underlying vaticanol C-induced apoptosis in HL-60 cells. Treatment of HL-60 cells with vaticanol C was found to cause a marked decrease in the level of phosphorylated extracellular signal-regulated kinase (ERK) concurrent with inhibited phosphorylation of its upstream kinase mitogen-activates protein kinase kinase (MEK). Moreover, exposure to vaticanol C led to a significant reduction in the level of phosphorylated Akt. Thus, vaticanol C induced inhibition of both ERK and Akt phosphorylation, resulting in reduced phosphorylation of Bad. These results suggest that vaticanol C might induce apoptosis via a mechanism involving activation of Bad through disruption of pro-survival signaling pathways.
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2-Hydroxyergothioneine, a New Ergothioneine Derivative from the Mushroom Lyophyllum connatum, and Its Protective Activity against Carbon Tetrachloride-Induced Injury in Primary Culture Hepatocytes
Chieko Kimura1, Manabu Nukina1,2, Kiharu Igarashi1,2, and Yoko Sugawara3
1Course of the Science of Bioresources, The United Graduate School of Agricultural Sciences, Iwate University (Yamagata University), 1-23 Wakaba-cho, Tsuruoka, Yamagata 997-8555, Japan
2Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, 1-23 Wakaba-cho, Tsuruoka, Yamagata 997-8555, Japan
3School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, JapanA new ergothioneine derivative named 2-hydroxyergothioneine was isolated from the mushroom Lyophyllum connatum. Ergothioneine,N-hydroxy-N,N-dimethylurea, and connatin (N-hydroxy-N,N-dimethylcitrulline) were also isolated. All the compounds displayed the ability to scavenge free radicals, based on a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Structural determination, including the absolute stereochemistry of 2-hydroxyergothioneine, was achieved by spectroscopic analysis and X-ray crystallography. The radical scavenging activity of 2-hydroxyergothioneine was almost the same as that of ergothioneine. 2-Hydroxyergothioneine showed the greatest protective activity against carbon tetrachloride-induced injury in primary culture hepatocytes.
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Gene Cloning, Overproduction, and Characterization of Thermolabile Alkaline Phosphatase from a Psychrotrophic Bacterium
Yutaka Suzuki1, Yoichi Mizutani1, Tadao Tsuji1, Naoto Ohtani1, Kazufumi Takano1,2, Mitsuru Haruki1, Masaaki Morikawa1, and Shigenori Kanaya1
1Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
2PRESTO, JST, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
The gene encoding alkaline phosphatase from the psychrotrophic bacterium Shewanella sp. SIB1 was cloned, sequenced, and overexpressed in Escherichia coli. The recombinant protein was purified and its enzymatic properties were compared with those of E. coli alkaline phosphatase (APase), which shows an amino acid sequence identity of 37%. The optimum temperature of SIB1 APase was 50 ℃, lower than that of E. coli APase by 30 ℃. The specific activity of SIB1 APase at 50 ℃ was 3.1 fold higher than that of E. coli APase at 80 ℃. SIB1 APase lost activity with a half-life of 3.9 min at 70 ℃, whereas E. coli APase lost activity with a half-life of >6 h even at 80 ℃. Thus SIB1 APase is well adapted to low temperatures. Comparison of the amino acid sequences of SIB1 and E. coli APases suggests that decreases in electrostatic interactions and number of disulfide bonds are responsible for the cold-adaptation of SIB1 APase.
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Enantioselective Total Synthesis of Enokipodins AD, Antimicrobial Sesquiterpenes Produced by the Mushroom, Flammulina velutipes
Mana Saito and Shigefumi Kuwahara
Laboratory of Applied Bioorganic Chemistry, Graduate School of Agricultural Science, Tohoku University, Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981-8555, Japan
The first enantioselective total synthesis of enokipodins A, B, C and D, highly oxidized ±-cuparenone-type sesquiterpenoids possessing antimicrobial activity, was accomplished in 828% overall yields from methyl (2,5-dimethoxy-4-methylphenyl)acetate by applying Meyers diastereoselective alkylation protocol for the construction of their C7-quaternary asymmetric center. The present synthesis confirmed the absolute configuration of the enokipodins, and also constitutes a formal enantioselective synthesis of (S)-1,4-cuparenediol and (S)-cuparene-1,4-quinone.
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Molecular Dissection of the Promoter of the Light-Induced and Circadian-Controlled APRR9 Gene Encoding a Clock-Associated Component of Arabidopsis thaliana
Shogo Ito, Norihito Nakamichi, Akinori Matsushika, Toru Fujimori, Takafumi Yamashino, and Takeshi Mizuno
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
In the model higher plant Arabidopsis thaliana, a number of circadian clock-associated protein components have recently been identified. Among them, a small family of ARABIDOPSIS PSEUDO-RESPONSE REGULATORS (APRR1/TOC1, APRR3, APRR5, APRR7, and APRR9) is interesting because the most probable clock component TIMING OF CAB EXPRESSION 1 (TOC1) belongs to this family. Several lines of evidence have already been accumulated to support the view that not only APRR1/TOC1 but also other APRR family members are crucial for a better understanding of the molecular link between circadian rhythm and light-signal transduction. Among the APRR1/TOC1 family members, the circadian-controlled APRR9 gene is unique in that its expression is rapidly induced by light at the level of transcription. In this study we dissected the regulatory cis-elements of the light-induced and/or circadian-controlled APRR9 promoter by employing not only a mutant plant carrying a T-DNA insertion in the APRR9 promoter, but also a series of APRR9-promoter::LUC (luciferase) reporters that were introduced into an Arabidopsis cultured cell line (T87 cells). Taking the results of these approaches together, we provide several lines of evidence that the APRR9 promoter contains at least two distinctive and separable regulatory cis-elements: an L element responsible for the light-induced expression, followed by an R element necessary for the fundamental rhythmic expression of APRR9. Furthermore, APRR1/TOC1 was implicated in the L-element-mediated light response of APRR9, directly or indirectly.
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Synthesis and Algicidal Activity of (+)-Cyanobacterin and Its Stereoisomer
Fumito Ishibashi1, Suhwan Park1, Takako Kusano1, and Kazuyoshi Kuwano2
1Faculty of Fisheries, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan
2Graduate School of Science and Technology, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan
(+)-Cyanobacterin, a photosynthesis inhibitor of freshwater cyanobacterium Schytonema hofmanni, was synthesized in 6 steps from a readily accessible chiral synthon 5R-5-(l-menthyloxy)-2(5H)furanone.
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Microstructures of Bread Dough and the Effects of Shortening on Frozen Dough
Shigeo Aibara1, Noriko Ogawa2, and Masaaki Hirose1
1Division of Applied Life Science, Graduate School of Agriculture, Kyoto University, Gokasho Uji, Kyoto 611-0011, Japan
2Faculty of Home Economics, Gifu Womens University, 80 Taromaru, Gifu 501-2529, JapanThree types of straight doughs different in combination of yeast and shortenings (RLS20, FTS20, and FTS80) were prepared, and the structure of the frozen doughs was examined under a microscope after staining protein or lipid droplets. Even after 2 months of frozen storage, distinct changes were not found in the gluten network of FTS80, although significant damages in the dough structures of FTS20 and RLS20 appeared after only one month of frozen storage. These results suggest that the gluten networks loosen and decrease in the water retention ability, and it may be concluded that the lipid is removed from the gluten protein due to the decrease in water in the continuous protein phase. The resulting product from the damage to the gluten matrix gave rise to fusion of lipid droplets and an increase in their size. Because of the difference in fatty acid composition, the lipids of shortening S80 are presumed to interact more strongly with gluten proteins and to keep the gluten matrix from damage in comparison with the lipids of shortening S20.
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Note
Sericin Enhances Attachment of Cultured Human Skin Fibroblasts
Kozo Tsubouchi, Yumiko Igarashi, Yoko Takasu, and Hiromi Yamada
National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan
Human skin fibroblasts were cultured on sericin prepared from cocoon shells. The living cell number after 72 h was enhanced to 250% of the no-sericin control. The increase was due to the acceleration of the initial attachment of the cells. It was found that sericin M, the main component of about 400 kDa, and its serine-rich repetitive domain were the active principles.
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Note
Cloning, Expression in Yeast, and Functional Characterization of CYP76A4, a Novel Cytochrome P450 of Petunia That Catalyzes ( É -1)-Hydroxylation of Lauric Acid
Katsutomo Tamaki1, Hiromasa Imaishi2, Hideo Ohkawa3, Kiyoharu Oono2, and Mikihiro Sugimoto1
1Laboratory of Biotechnology, Hyogo Prefectural Research Institute for Agriculture, Minaminooka-ko 1533, Befu-cho, Kasai-shi, Hyogo 679-0198, Japan
2Functional Analysis of Environmental Genes, Research Center for Environmental Genomics, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan
3Department of Applied Biological Science, Faculty of Life Science, Fukuyama University, Gakuen-cho 1, Fukuyama, Hiroshima 720-0292, JapanA cDNA clone of a novel cytochrome P450, CYP76A4, was isolated from Petunia hybrida. The cDNA clone contained an open reading frame (ORF) encoding a predicted 510 amino acid polypeptide. The CYP76A4 cDNA was expressed in yeast Saccharomyces cerevisiae AH22. Recombinant yeast microsomes containing the CYP76A4 hemoprotein were found to catalyze ( É -1)-hydroxylation of lauric acid.
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Note
Circadian-Associated Rice Pseudo Response Regulators (OsPRRs): Insight into the Control of Flowering Time
Masaya Murakami1, Akinori Matsushika1, Motoyuki Ashikari2, Takafumi Yamashino1, and Takeshi Mizuno1
1Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
2Bioscience and Biotechnology Center, Nagoya University, Chikusa-ku, Nagoya 464-8601, JapanA small family of plant proteins, designated PSEUDO RESPONSE REGULATORS (PRRs), is crucial for a better understanding of the molecular link between circadian rhythm and photoperiodic control of flowering time in the dicotyledonous model plant Arabidopsis thaliana. Recently, we showed that the monocotyledonous model plant Oryza sativa also has homologous members of the OsPRR family (Oryza sativa PRR). In the previous experiments with rice, we mainly characterized a japonica variety (Nipponbare). By employing an indica variety (Kasalath), in this study we further characterized OsPRRs with reference to the photoperiod sensitivity Hd (Heading date) QTL (quantitative trait loci) implicated in the control of flowering time in rice. The circadian-controlled and sequential expression profiles of the five OsPRR genes were observed not only for Nipponbare but also for Kasalath. Then each of these OsPRR genes was mapped on the rice chromosomes. Among these OsPRR genes, OsPRR37 was mapped very closely to Hd2-QTL, which was identified as the major locus that enhances the photoperiod sensitivity of flowering in Nipponbare. Furthermore, we found that Kasalath has a severe mutational lesion in the OsPRR37 coding sequence.
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Note
Transient RNAi Induction against Endogenous Genes in Arabidopsis Protoplasts Using in Vitro-Prepared Double-Stranded RNA
Chung-Il An, Aki Sawada, Yoshiaki Kawaguchi, Ei-ichiro Fukusaki, and Akio Kobayashi
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
RNA interference is a powerful technique for suppressing gene functions in many eukaryotes including plants. Here we show that introduction of double-stranded RNA into Arabidopsis protoplasts leads to marked silencing of endogenous genes, as observed previously for transgenes [Biosci. Biotechnol. Biochem., 67, 26742677 (2003)]. This simple system should be useful for functional analysis of genes involved in fundamental cellular processes.
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Note
Cold-Active Polygalacturonase from Psychrophilic-Basidiomycetous Yeast Cystofilobasidium capitatum Strain PPY-1
Tomoyuki Nakagawa, Toshinori Nagaoka, Tatsuro Miyaji, and Noboru Tomizuka
Department of Food Science and Technology, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido 099-2493, Japan
We purified and characterized a cold-active polygalacturonase (PG) from the extracellular fraction of Cystofilobasidium capitatum strain PPY-1. The purified PG from strain PPY-1 has a molecular mass of about 44 kDa, and exhibited high activity at 0 ℃, although its optimum temperature was 45 ℃. Although the Km value for polygalacturonate as a substrate at 45 ℃ was found to be 11.2 mg/ml, it decreased gradually with decreasing temperature, and it was 0.66 mg/ml at 0 ℃. Moreover, its cleavage pattern was of the endo-type. These findings might indicate that PG from strain PPY-1 is a novel type of cold-active endo-PG that is able to degrade pectin compounds at low temperatures.
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Note
Structural and Functional Analysis of pTB6 from Bifidobacterium longum
Kunihito Tanaka, Kazuya Samura, and Yasunobu Kano
Department of Molecular Genetics, Institute of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, 1 Shichono-cho, Misasagi, Yamashina-ku, Kyoto 607-8412, Japan
The complete nucleotide sequence for pTB6 (3,624 bp) from Bifidobacterium longum was determined. This plasmid is 95% homologous in nucleotide (nuc) sequence, and also 92% in RepB aa sequence, to rolling circle replication (RCR) plasmids pKJ36 and pB44, suggesting that pTB6 replicates by the rolling circle mechanism. The putative MembB, MobA, and protein encoding from orf (Orf) I detected were nonessential for plasmid replication. We constructed an immobile shuttle vector from pTB6 and pUC18, which transformed B. longum with a high efficiency of 2.5 × 106 transformants/¼g DNA.
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Note
PRR5 (PSEUDO-RESPONSE REGULATOR 5) Plays Antagonistic Roles to CCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) in Arabidopsis thaliana
Toru Fujimori, Eriko Sato, Takafumi Yamashino, and Takeshi Mizuno
Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
In general, the clock (or oscillator) is central to circadian rhythms in many organisms. In the model higher plant Arabidopsis thaliana, the best candidates forclock components are CCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) and LHY (LATE ELONGATED HYPOCOTYL), which are homologous Myb-related transcription factors. It is also believed that TOC1 (TIMING OF CAB EXPRESSION 1) is another component of the central oscillator. In this connection, we have been characterizing a small family of proteins, designated ARABIDOPSIS PSEUDO-RESPONSE REGULATOR (PRR1, PRR3, PRR5, PRR7, and PRR9), based on the fact that one of the members (PRR1) is identical to TOC1. Nevertheless, it is not yet certain whether other PRR family members are also implicated in clock function per se. To address this issue, in this study we examined a functional interaction between the CCA1 clock component and one of the PRR family members, PRR5, by employing transgenic lines overexpressing both the CCA1 and PRR5 genes. Evidence will be provided that PRR5 plays an antagonistic role(s) to the putative CCA1 clock component.