(Vol.66 No.8 2002)
Review
Mevalonate and Nonmevalonate Pathways for the Biosynthesis of Isoprene Units
Tomohisa KUZUYAMA p.1619
Bioavailability of a Series of Novel Acylated Ascorbic Acid Derivatives,
6-O-Acyl-2-O-α-D-glucopyranosyl-L-ascorbic Acids, as an Ascorbic Acid Supplement
in Rats and Guinea Pigs
Akihiro TAI, Yoshihito FUJINAMI, Kyoko MATSUMOTO, Daisuke KAWASAKI, and Itaru
YAMAMOTO p.1628
Heating of an Ovalbumin Solution at Neutral pH and High Temperature
Songsin PHOTCHANACHAI, Alka MEHTA, and Naofumi KITABATAKE p.1635
Generation of Reactive Oxygen Species and Photon Emission from
a Browned Product
Tetsuo IIDA,, Yumiko YOSHIKI, Shinich SOMEYA, and Kazuyoshi OKUBO p.1641
Isolation from Nocardioides sp. Strain CT16, Purification, and
Characterization of a Deoxycytidine Deaminase Extremely Thermostable in the
Presence of D,L-Dithiothreitol
Tsuyoshi SAKAI, Takahiro DAIKAI, Hideo MONMA, and Hidekatsu MAEDA p.1646
Changes in Flower Coloration and Sepal Anthocyanins of Cyanic
Delphinium Cultivars during Flowering
Fumio HASHIMOTO, Mika TANAKA,＃ Hiroko MAEDA, Shoko FUKUDA, Keiichi SHIMIZU,
and Yusuke SAKATA p.1652
Functional Analyses of Tyr420 and Leu607 of Alicyclobacillus acidocaldarius
Squalene-Hopene Cyclase. Neoachillapentaene, a Novel Triterpene with the 1,5,6-Trimethylcyclohexene
Moiety Produced through Folding of the Constrained Boat Structure
Tsutomu SATO, Shigehiro SASAHARA, Toshiyuki YAMAKAMI, and Tsutomu HOSHINO p.1660
Bleaching Activity of 4-Phenyl-3-(substituted benzylthio)-4H-1,2,4-triazoles
Naotaka YAMADA, Daisuke KUSANO, and Eiich KUWANO p.1671
Effects of Acetan on Production of Bacterial Cellulose by Acetobacter
xylinum
Takehiko ISHIDA, Yasushi SUGANO, Tomonori NAKAI, and Makoto SHODA p.1677
Primary Structure and Catalytic Properties of a Cold-active Esterase
from a Psychrotroph, Acinetobacter sp. Strain No. 6. Isolated from Siberian
Soil
Takeshi SUZUKI,1 Toru NAKAYAMA,2 Tatsuo KURIHARA,1 Tokuzo NISHINO,2 and
Nobuyoshi ESAKI1, p.1682
Genomic Structure and Promoter Analysis of Phosphoenolpyruvate
Carboxylase in a C3 Plant, Nicotiana sylvestris
Nozomu KOIZUMI,1,3, Kentaro TOYOTA,1, Sakihito KITAJIMA,1 Yasuyuki YAMADA,1,3
and Fumihiko SATO1,2, p.1691
New Convulsive Compounds, Brasiliamides A and B, from Penicillium
brasilianum Batista JV-379
Tomoyuki FUJITA, Daisuke MAKISHIMA, Kohki AKIYAMA, and Hideo HAYASHI p.1697
Inhibitory Properties of Antitumor Prostaglandins against Topoisomerases
Keitarou SUZUKI and Masaru UYEDA p.1706
Effects of Dietary Medium-Chain Triacylglycerols on Serum Lipoproteins
and Biochemical Parameters in Healthy Men
Naohisa NOSAKA, Michio KASAI, Masahiro NAKAMURA, Isamu TAKAHASHI, Megumi ITAKURA,
Hiroyuki TAKEUCHI, Toshiaki AOYAMA, Hiroaki TSUJI, Mitsuko OKAZAKI, and Kazuo
KONDO
p.1713
Determination of the Absolute Configurations of the Anteiso Acid
Moieties of Glycoglycerolipid S365A Isolated from Corynebacterium aquaticum
Kazuaki AKASAKA, Seiya SHICHIJYUKARI, Hiroshi MEGURO, and Hiroshi OHRUI p.1719
Solubility of Saturated Fatty Acids in Water at Elevated Temperatures
Pramote KHUWIJITJARU, Shuji ADACHI, and Ryuichi MATSUNO p.1723
Note
Isolation of Antimicrobial Compounds from Guava (Psidium guajava L.) and their
Structural Elucidation
Hidetoshi ARIMA and Gen-ichi DANNO p.1727
Note
Potent Limonoid Insect Antifeedant from Melia azedarach
Cecilia CARPINELLA,1 Carlos FERRAYOLI,1 Graciela VALLADARES,2 Maria DEFAGO,2
and Sara PALACIOS1 p.1731
Note
Sex Differences in Kidney Mineral Concentrations and Urinary Albumin Excretion
in Rats Given High-phosphorus Feed
Hiroshi MATSUZAKI,1 Shin-ichi KATSUMATA,2 Ritsuko MASUYAMA,2 Mariko UEHARA,2
Kahoru NAKAMURA,1 and Kazuharu SUZUKI2 p.1737
Note
Alternative Splicing Products of the Gene for a Human Nuclear Actin-related
Protein, hArpNβ/Baf53, that Encode a Protein Isoform, hArpNβS, in the Cytoplasm
Eri OHFUCHI, Katsuhiko NISHIMORI, and Masahiko HARATA p.1740
Note
Iron Absorption in Rats Increased by Yeast Glucan
Ta Thi Tuyet MAI, Kaori IGARASHI, Rieko HIRUNUMA, Satoko TAKASAKI, Masaaki YASUE,
Shuichi ENOMOTO, Shuichi KIMURA, and Nguyen Van CHUYEN p.1744
Note
Shogaols from Zingiber officinale as Promising Antifouling Agents
Hideo ETOH, Takeyoshi KONDOH, Rikoh NODA, Inder Pal SINGH, Yohko SEKIWA,
Kohjiro MORIMITSU, and Kikue KUBOTA p.1748
Note
Composition of Seed Storage Proteins Changed by Glutathione Treatment of Soybeans
Motoko AWAZUHARA,1,3 Hoyeun KIM,1,2 Hiroaki HAYASHI,1 Mitsuo CHINO,1,4 Sang-Gu
KIM,3 and Toru FUJIWARA1, P.1751
Note
Improving Effect of Dietary Taurine Supplementation on the Oxidative Stress
and Lipid Levels in the Plasma, Liver and Aorta of Rabbits Fed on a High-Cholesterol
Diet
Jale BALKAN,1, <Oh><0196><Wa>Oznur KANBA<Oh><0188><Wa>GLI,1
Aydan HAT<Oh>_<Wa>IPO<Oh><0188><Wa>GLU,1 Mutlu
K<Oh><0196><Wa>U<Oh><0199><0196><Wa>UK,2
U<Oh><0178><Wa>gur <Oh><0199><Wa>EV<Oh>_<Wa>IKBAflS,3
Gul<Oh><0231><Wa>in AYKA<Oh><0199><Wa>-TOKER,1
and M<Oh><0228><Wa>ujdat UYSAL1 P.1755
Note
Effects of Lentinus edodes on Fatty Acid and Molecular Species Profiles of Phosphatidylcholine
in Rats Fed Different Levels of Corn Oil
Yasuhiko SHIMADA, Tatsuya MORITA, and Kimio SUGIYAMA P.1759
Note
Synthetic Studies of Proanthocyanidins. Highly Stereoselective Synthesis of
the Catechin Dimer, Procyanidin-B3
Akiko SAITO,1 Noriyuki NAKAJIMA,2 Akira TANAKA,3 and Makoto UBUKATA2, P.1764
Note
Tocotrienol Levels in Various Tissues of Sprague-Dawley Rats after Intragastric
Administration of Tocotrienols
Masaaki OKABE, Mariko OJI, Ikuo IKEDA, Hirofumi TACHIBANA, and Koji YAMADA
P.1768
Note
Lipase-Catalyzed Kinetic Resolution of (±)-cis-Flavan-4-ol and Its Acetate:
Synthesis of Chiral 3-Hydroxyflavanones
Tamotsu TODOROKI,1 Akiko SAITO,2 and Akira TANAKA1, P.1772
Note
Purification and Properties of a Carbonyl Reductase Useful for Production of
Ethyl (S)-4-Chloro-3-hydroxybutanoate from Kluyveromyces lactis
Hiroaki YAMAMOTO, Norihiro KIMOTO, Akinobu MATSUYAMA, and Yoshinori KOBAYASHI
P.1775
Note
Determination of the Stereochemistry of (－)-Koninginin A by an X-ray Analysis
of Its Synthetic Sample
Kenji MORI,1,‡ Masahiko BANDO,2 and Kaoru ABE3 P.1779
-1-
Review
Mevalonate and Nonmevalonate Pathways for the Biosynthesis of Isoprene Units
Tomohisa KUZUYAMA
Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan
Isoprenoids are synthesized by consecutive condensations of their five-carbon precursor, isopentenyl diphosphate, to its isomer, dimethylallyl diphosphate. Two pathways for these precursors are known. One is the mevalonate pathway, which operates in eucaryotes, archaebacteria, and cytosols of higher plants. The other is a recently discovered pathway, the nonmevalonate pathway, which is used by many eubacteria, green algae, and chloroplasts of higher plants. To date, five reaction steps in this new pathway and their corresponding enzymes have been identified. EC numbers of these enzymes have been assigned by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC-IUBMB) and are available at http://www.chem.qmw.ac.uk/iubmb/enzyme/reaction/terp/nonMVA.html.
Key words: isoprenoid; biosynthesis; mevalonate pathway; nonmevalonate pathway; inhibitor
-2-
Bioavailability of a Series of Novel Acylated Ascorbic Acid Derivatives, 6-O-Acyl-2-O-α-D-glucopyranosyl-L-ascorbic
Acids, as an Ascorbic Acid Supplement in Rats and Guinea Pigs
Akihiro TAI, Yoshihito FUJINAMI, Kyoko MATSUMOTO, Daisuke KAWASAKI, and Itaru YAMAMOTO
Department of Immunochemistry, Faculty of Pharmaceutical Sciences, Okayama University, Okayama 700-8530, Japan
Received November 5, 2001; Accepted April 5, 2002
The bioavailability of a series of novel acylated ascorbic acid derivatives, 6-O-acyl-2-O-α-D-glucopyranosyl-L-ascorbic acids (6-Acyl-AA-2G), as an ascorbic acid (AA) supplement was investigated in rats and guinea pigs. Oral administration of 6-Acyl-AA-2G to rats resulted in an increase in the plasma AA level. However, the intact form was not detectable in the plasma by high-performance liquid chromatography, indicating its hydrolysis through the process of absorption. After an intravenous injection to rats of 6-Octa-AA-2G as a representative derivative, the intact form rapidly disappeared from the plasma, being followed by a prolonged and marked elevation of the plasma AA level. Various tissue homogenates from guinea pigs were examined for their releasing activity of AA, 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G) and 6-O-acyl-AA from 6-Acyl-AA-2G. High activity was observed in the small intestine. These hydrolytic activities to AA and 6-O-acyl-AA were completely inhibited by castanospermine, an α-glucosidase inhibitor, and AA-2G was observed as the only resulting hydrolysate, suggesting the participation of α-glucosidase and esterase in the in vivo hydrolysis of 6-Acyl-AA-2G. 6-Octa-AA-2G was found to exhibit an obvious therapeutic effect in scorbutic guinea pigs from its repeated oral administration. These results indicate that 6-Acyl-AA-2G is a readily available source of AA activity in vivo, and may be useful as an effective pharmacological agent and as a promising food additive.
Key words: 6-Acyl-AA-2G; lipophilic ascorbate; stable ascorbate; antiscorbutic activity; metabolism
-3-
Heating of an Ovalbumin Solution at Neutral pH and High Temperature
Songsin PHOTCHANACHAI, Alka MEHTA, and Naofumi KITABATAKE
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan
Received November 8, 2001; Accepted April 9, 2002
The thermal denaturation, aggregation, and degradation of hen egg white ovalbumin dissolved in distilled and deionized water (60 mg/ml, pH 7.5) was investigated by differential scanning calorimetry (DSC), polyacrylamide gel electrophoresis (PAGE), and viscosity measurement. Two independent endothermic peaks were observed up to 180°C by the DSC analysis. The first peak appeared at around 80°C, corresponding to the denaturation temperature of ovalbumin. The second peak occurred around 140°C due to the degradation of protein molecules as judged from the analysis by SDS-PAGE. The viscosity of the ovalbumin solution increased dramatically above 88°C and maintained almost the same value up until heating to 140°C. The increase in viscosity after heating to 88°C was due to the denaturation and subsequent aggregation of ovalbumin molecules as observed by SDS-PAGE. The decrease in viscosity of the samples heated above 150°C appears to have been the result of degradation of the ovalbumin molecules.
Key words: egg white protein; ovalbumin; molecular aggregation; degradation; high-temperature heating
-4-
Generation of Reactive Oxygen Species and Photon Emission from a Browned Product
Tetsuo IIDA,, Yumiko YOSHIKI, Shinich SOMEYA, and Kazuyoshi OKUBO
Department of Molecular Bioscience, Graduate School of Life Science, Tohoku University, 1-1 Tsutsumidori Amamiyamachi, Aoba-ku, Sendai, Miyagi 981-8555, Japan
Received December 17, 2001; Accepted April 16, 2002
The properties of photon emission arising from a browned product were investigated. The photon intensity of the browned product was proportional to the absorbancy at 420 nm, and was influenced by the amino acid structure. The fluorescence spectrum showed similar compounds in the browned product to be related with this photon emission. Superoxide and hydrogen peroxide contributed highly to this photon emission, and several redox compounds enhanced the photon intensity at appropriate concentrations. Our work suggests that the photon intensity was closely related to the reactive oxygen species (ROS) generated from the browned product, and this effect may be utilized to evaluate the function and quality of browned food.
Key words: browned product; soy sauce; coffee; photon emission; reactive oxygen species
-5-
Isolation from Nocardioides sp. Strain CT16, Purification, and Characterization
of a Deoxycytidine Deaminase Extremely Thermostable in the Presence of D,L-Dithiothreitol
Tsuyoshi SAKAI, Takahiro DAIKAI, Hideo MONMA, and Hidekatsu MAEDA
Department of Bioengineering, Faculty of Engineering, Soka University, 1-236, Tangi, Hachioji, Tokyo 192-8577, Japan
Received December 19, 2001; Accepted April 9, 2002
A deoxycytidine deaminase that was extremely thermostable in the presence of dithiothreitol was found in a mesophilic bacterium isolated from soil. The bacterium was classified as a Nocardioides sp. The enzyme was purified to a homogeneous protein by treatment at 100°C, ammonium sulfate precipitation, and chromatography on DEAE-Toyopearl, hydroxyapatite, and then Sephacryl S-100. Twenty micrograms of the pure enzyme was obtained from 811 mg of the starting crude protein. After treatment at 50°C for 15 min in the absence of dithiothreitol, enzyme activity was 44％ of the starting activity; after treatment at 100°C for 2 h in the presence of 50 mM dithiothreitol, activity was 56％ of the starting activity. Dithiothreitol greatly stabilized the enzyme. Activity was maximum at 99°C. The Km values for deoxycytidine, cytidine, and methyl-deoxycytidine were 55.2 , 140, and 130 μM, respectively. The molecular mass was estimated to be 52 kDa by gel permeation chromatography. The enzyme molecule was dissociated into two subunits each of 18 kDa subunit when reduced with mercaptoethanol.
Key words: thermostable; deoxycytidine deaminase; Nocardioides; dithiothreitol; mesophilic bacteria
-6-
Changes in Flower Coloration and Sepal Anthocyanins of Cyanic Delphinium Cultivars
during Flowering
Fumio HASHIMOTO, Mika TANAKA,＃ Hiroko MAEDA, Shoko FUKUDA, Keiichi SHIMIZU, and Yusuke SAKATA
Ornamental Horticulture Laboratory, Department of Horticultural Science, Faculty of Agriculture, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-0065, Japan
Received December 21, 2001; Accepted April 18, 2002
The changes in flower color related to sepal pigmentation of cyanic Delphinium cultivars were investigated during anthesis. The sepal hues of the purple and blue flowered varieties observed on the initial day of unfurling had changed with a decrease in hue angle three days after anthesis. In both the purple and blue cultivars, violdelphin (3) was the major component on day one of anthesis, and the chromaticity improved with increasing sepal concentrations of violdelphin (3) and cyanodelphin (4) after three days of unfurling. The flower hue was dominated by the constitution of acylated anthocyanins, and the chromaticity was ordered by the sepal concentration. The biosynthesis of cyanodelphin (4) from violdelphin (3) was postulated since an increase in the sepal concentration of cyanodelphin (4) was accompanied by a decrease in violdelphin (3). Acylation of the anthocyanins was initiated by an increase in the respective possible precursors, tulipanin (2) and violdelphin (3), to subsequently synthesize violdelphin (3) and cyanodelphin (4) during flowering.
Key words: biogenesis; cyanodelphin; Delphinium; Ranunculaceae; violdelphin
-7-
Functional Analyses of Tyr420 and Leu607 of Alicyclobacillus acidocaldarius
Squalene-Hopene Cyclase. Neoachillapentaene, a Novel Triterpene with the 1,5,6-Trimethylcyclohexene
Moiety Produced through Folding of the Constrained Boat Structure
Tsutomu SATO, Shigehiro SASAHARA, Toshiyuki YAMAKAMI, and Tsutomu HOSHINO
Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Ikarashi, Niigata 950-2181, Japan
Received January 10, 2002; Accepted March 27, 2002
The functions of Tyr420 and Leu607 were analyzed by constructing various site-directed mutants. The mutation at position 420 into Ala and Gly gave bicyclic α- and γ-polypodatetraene in significant amounts, but with a trace amount of tricyclic malabaricatriene. The kinetic data for and the product distribution of the Y420F mutant indicate that the major function of Tyr420 is to stabilize the 6/6-fused bicyclic cation. Mutation experiments on Leu607 demonstrate that the appropriate steric bulk size at position 607 is required to strongly bind with the product-like conformation formed during the polycyclization process. Introduction of the bulkiest Trp residue into 420 or 607 led to the production of a novel monocyclic triterpene having the (5R,6R)-1,5,6-trimethylcyclohexene ring, named neoachillapentaene, indicating that the enzymatic cyclization proceeded via a constrained boat structure. Folding of the squalene molecule into a boat conformation by squalene cyclase has not been reported before.
Key words: squalene; hopene; terpene cyclase; unnatural natural product; Alicyclobacillus acidocaldarius
-8-
Bleaching Activity of 4-Phenyl-3-(substituted benzylthio)-4H-1,2,4-triazoles
Naotaka YAMADA, Daisuke KUSANO, and Eiich KUWANO
Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan
Received January 30, 2002; Accepted March 23, 2002
A variety of 4-aryl- and 4-alkyl-3-(substituted benzylthio)-4H-1,2,4-triazoles were prepared and evaluated for their bleaching activity by the lettuce seedling test.
Among the series of tested compounds, 4-(3-fluorophenyl)-3-(4-trifluoromethylbenzylthio)-4H-1,2,4-triazole (39) exhibited the highest bleaching activity, causing complete bleaching symptoms at 10 μM. In the dark
condition, compound 39 inhibited the formation of such carotenoids as β-carotene, violaxanthin, neoxanthin and lutein, resulting in the formation of ζ-carotene, phytoene, phytofluene and β-zeacarotene, which were not detected in the untreated control. Treatment by compound 39 at 50 μM resulted in the amount of accumulated ζ-carotene being seven-fold higher than that of phytoene, phytofluene and β-zeacarotene. These results suggest that compound 39 might have interfered with desaturation, especially ζ-carotene desaturation, during carotenoid biosynthesis.
Key words: bleaching; ζ-carotene; β-carotene; triazole
-9-
Effects of Acetan on Production of Bacterial Cellulose by Acetobacter xylinum
Takehiko ISHIDA, Yasushi SUGANO, Tomonori NAKAI, and Makoto SHODA
Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8503, Japan
Received February 4, 2002; Accepted March 20, 2002
Acetan is a water-soluble polysaccharide produced by a bacterial cellulose (BC) producer, Acetobacter xylinum. An acetan-nonproducing mutant, EP1, was generated from wild-type A. xylinum BPR2001 by the disruption of aceA, which may act to catalyze the first step of the acetan biosynthetic pathway in this bacterium. EP1 produced less BC than the wild-type strain. However, when EP1 was cultured in a medium containing acetan, BC production was stimulated and the final yield of BC was equivalent to that of BPR2001. The culture broth containing acetan was more viscous and the free cell number was higher than that of the broth without the polysaccharide, so acetan may hinder the coagulation of BC in the broth. The addition of 1.5 g/l agar also increased BC production; we concluded that acetan and BC syntheses were not directly related on the genetic level.
Key words: Acetobacter xylinum; bacterial cellulose; acetan; viscosity
-10-
Primary Structure and Catalytic Properties of a Cold-active Esterase from a
Psychrotroph, Acinetobacter sp. Strain No. 6. Isolated from Siberian Soil
Takeshi SUZUKI,1 Toru NAKAYAMA,2 Tatsuo KURIHARA,1 Tokuzo NISHINO,2 and Nobuyoshi ESAKI1,
1Laboratory of Microbial Biochemistry, Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan 2Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba-yama 07, Sendai, Miyagi 980-8579, Japan
Received February 6, 2002; Accepted March 27, 2002
We cloned a gene coding for a cold-active esterase from a genomic library of Acinetobacter sp. strain No. 6, a psychrotroph isolated from Siberian soil. The gene, aest, encoded a protein of 301 amino acid residues, the deduced sequence of which had less than 17％ identity to sequences of known esterases and lipases. However, the esterase seemed to belong to the α/β hydrolase superfamily, because it contained a sequence, Gly-Xaa-Ser-Xaa-Gly (with Xaa an arbitrary amino acid residue), found in most serine hydrolases of this superfamily. Sequence comparison earlier suggested a weak phylogenetic relationship of gene product AEST to the EST group of the esterase-lipase family, which has been found only in eukaryotes. The aest gene was expressed in Escherichia coli BL21(DE3) cells under the control of the T7 promoter, and the expression product was purified to homogeneity and characterized. It catalyzed the hydrolysis of esters with short-chain acyl groups and had lower activation energy and lower thermostability than do mesophilic enzymes, as expected from the cold-adapted nature of this enzyme.
Key words: cold-active; esterase; prokaryotic; psychrotroph; superfamily
-11-
Genomic Structure and Promoter Analysis of Phosphoenolpyruvate Carboxylase in
a C3 Plant, Nicotiana sylvestris
Nozomu KOIZUMI,1,3, Kentaro TOYOTA,1, Sakihito KITAJIMA,1 Yasuyuki YAMADA,1,3 and Fumihiko SATO1,2,
1Division of Applied Life Sciences, Graduate School of Agriculture, 2Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Sakyo, Kyoto 606-8502, Japan 3Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Takayama, Ikoma, Nara 630-0101, Japan
Received February 12, 2002; Accepted April 4, 2002
Three genes encoding phosphoenolpyruvate carboxylase were isolated from Nicotiana sylvestris and designated Nsppc1--3. Sequencing of nucleotides showed that the coding sequence and deduced amino acid sequences were highly conserved among the genes, but sequences for noncoding regions including introns and 5′-flanking regions were not conserved. Analysis of the transcript level of the genes by a combination of reverse transcriptase-PCR and restriction fragment polymorphism showed mostNsppc1 in the leaves, stems, roots, and cultured cells of N. sylvestris. β-Glucuronidase activity was detected histochemically in mesophyll cells in leaves, lateral buds, and vascular bundles in roots of transgenic tobacco harboring a chimeric construct of the Nsppc1 promoter and the gene for β-glucuronidase. Deletion analysis indicated the presence of a silencer-like element for basal expression in the promoter region of Nsppc1.
Key words: phosphoenolpyruvate carboxylase; Nicotiana sylvestris; gene expression; genome structure
-12-
New Convulsive Compounds, Brasiliamides A and B, from Penicillium brasilianum
Batista JV-379
Tomoyuki FUJITA, Daisuke MAKISHIMA, Kohki AKIYAMA, and Hideo HAYASHI
Division of Applied Biological Chemistry, Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599--8531, Japan
Received February 14, 2002; Accepted May 1, 2002
New convulsive compounds, brasiliamides A (1) and B (2), were isolated by activity-guided fractionation from okara fermented with a soil isolate of Penicillium brasilianum Batista JV-379. Their structures were elucidated on the basis of spectral and chemical evidence and by X-ray crystallography of the hydrogenated product of 2. In the 1H- and 13C-NMR spectra of 2, the signals were complicated, all being doubled or broadened in several deuterated solvents at room temperature. The conformational change of 2 was clarified as the rotational isomerization of amide bonds in solution by NMR measurements at various temperatures. Four rotamers of 2 at two amide bonds were presented at －60°C in CDCl3, whereas only two isomers were apparent at room temperature, owing to rapid rotation of one of the amide bonds. Brasiliamides A and B respectively showed convulsive activity against silkworms with ED50 values of 300 and 50 μg/g of diet.
Key words: Penicillium brasilianum Batista; brasiliamide; rotational isomerization; convulsive activity; okara
-13-
Inhibitory Properties of Antitumor Prostaglandins against Topoisomerases
Keitarou SUZUKI and Masaru UYEDA
Laboratory of Pharmaceutical Microbiology, Faculty of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan
Received February 22, 2002; Accepted March 27, 2002
Prostaglandins (PGs) having antitumor activity such as Δ12,14-PGJ2, Δ12-PGJ2, PGA2 and PGA1 strongly inhibited topoisomerase II (topo II) from human placenta, the potential order of inhibitory activity of the PGs resembling that of the antitumor activity. PGs having no antitumor activity did not inhibit topo II. Δ12,14-PGJ2 to be a potent inhibitor showed inhibitions to some extent against topo I from wheat germ, NIH3T3 and calf thymus gland, and showed no inhibition against the enzymes from Vero, A549, HeLa and COLO 201 cells. Δ12,14-PGJ2 differentially inhibited topo I from different sources. Δ12,14-PGJ2 was a topo inhibitor of the cleavable complex-nonforming type without DNA intercalation.
Key words: prostaglandin; topoisomerase; antitumor; inhibition
-14-
Effects of Dietary Medium-Chain Triacylglycerols on Serum Lipoproteins and Biochemical
Parameters in Healthy Men
Naohisa NOSAKA, Michio KASAI, Masahiro NAKAMURA, Isamu TAKAHASHI, Megumi ITAKURA, Hiroyuki TAKEUCHI, Toshiaki AOYAMA, Hiroaki TSUJI, Mitsuko OKAZAKI, and Kazuo KONDO
Division of Healthcare Science Research Laboratory, Nisshin Oil Mills, Ltd., Kanagawa 239-0832, Japan Kagawa Nutrition University, Saitama 350-0288, Japan Institute of Environmental Science for Human Life, Ochanomizu University, Tokyo 112-8610, Japan
Received February 28, 2002; Accepted April 7, 2002
The objective of this study was to investigate effects of dietary medium-chain triacylglycerols (MCTs) on serum lipid levels, liver function, and hepatic fat accumulations in healthy men. Eleven subjects consumed 2200--2600 kcal daily, of which 70--80 g was fat; the fat included 40 g of MCTs or else 40 g of long-chain triacylglycerols (blended vegetable oil). The diet was followed for 4 weeks in this controlled double-blind study. At the end of the experiment, significant differences were not found in the concentrations of serum total cholesterol, very low density lipoprotein cholesterol, low density lipoprotein cholesterol, and high density lipoprotein cholesterol between the groups. Serum triglycerol levels were not significantly different in the groups. Adverse effects from ingestion of MCTs on liver functions, the liver-to-spleen ratio on computed tomography (an index of fatty liver), or results of blood tests were not seen. The results suggest that the long-term effects of dietary MCTs on serum cholesterol were similar to those of unsaturated fatty acids found abundantly in vegetable oil, and that consumption of MCTs in the amount of 40 g/day for a month does not cause liver fat accumulation or liver dysfunction.
Key words: medium-chain triacylglycerols; serum lipids; fatty liver; liver function; human
-15-
Determination of the Absolute Configurations of the Anteiso Acid Moieties of
Glycoglycerolipid S365A Isolated from Corynebacterium aquaticum
Kazuaki AKASAKA, Seiya SHICHIJYUKARI, Hiroshi MEGURO, and Hiroshi OHRUI
Graduate School of Life Sciences, Tohoku University, Tsutsumidori-Amamiyamachi 1-1, Aobaku, Sendai 981-8555, Japan, Tohoku Fukushi University, Kunimi, Aobaku, Sendai 981-0943, Japan
Received March 4, 2002; Accepted March 19, 2002
The absolute configurations of the two acid moieties, 12-methyltetradecanoate and 14-methylhexadecanoate, of glycoglycerolipid S365A isolated from Corynebacterium aquaticum were determined by an HPLC analysis after their conversion with the chiral fluorescent labeling reagents, (1S,2S)- and (1R,2R)-2-(2,3-anthracenedicarboximido)cyclohexanol. Both anteiso acids had the S configuration.
Key words: branched-chain fatty acid; chiral conversion; high-performance liquid chromatography; absolute configuration
-16-
Solubility of Saturated Fatty Acids in Water at Elevated Temperatures
Pramote KHUWIJITJARU, Shuji ADACHI, and Ryuichi MATSUNO
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
Received March 20, 2002; Accepted April 25, 2002
The solubility in water of saturated fatty acids with even carbon numbers from 8 to 18 was measured in the temperature range of 60 to 230°C and at a pressure of 5 or 15 MPa. The pressure had no significant effect on the solubility. The solubility of the fatty acids increased with increasing temperature. At temperatures higher than about 160°C, the logarithm of the solubility in mole fraction was linearly related to the reciprocal of the absolute temperature for each fatty acid, indicating that the water containing solubilized fatty acid molecules formed a regular solution at the higher temperatures. The enthalpy of a solution of the fatty acids in water, which was evaluated from the linear relationship at the given temperatures, increased linearly with the carbon number of the fatty acid
Key words: subcritical water; fatty acid; solubility; high temperature
-17-
Note
Isolation of Antimicrobial Compounds from Guava (Psidium guajava L.) and their
Structural Elucidation
Hidetoshi ARIMA and Gen-ichi DANNO
Division of Life Science, Graduate School of Science and Technology, Kobe University, Kobe 657-8501, Japan
Received November 5, 2001; Accepted April 10, 2002
Four antibacterial compounds were isolated from leaves of guava (Psidium guajava L.), and the structures of these compounds were established on the basis of chemical and spectroscopic evidence. Two new flavonoid glycosides, morin-3-O-α-L-lyxopyranoside and morin-3-O-α-L-arabopyranoside, and two known flavonoids, guaijavarin and quercetin, were identified. The minimum inhibition concentration of morin-3-O-α-L-lyxopyranoside and morin-3-O-α-L-arabopyranoside was 200 μg/ml for each against Salmonella enteritidis, and 250 μg/ml and 300 μg/ml against Bacillus cereus, respectively.
Key words: Psidium guajava; morin glycoside; Bacillus cereus; Salmonella enteritidis
-18-
Note
Potent Limonoid Insect Antifeedant from Melia azedarach
Cecilia CARPINELLA,1 Carlos FERRAYOLI,1 Graciela VALLADARES,2 Maria DEFAGO,2 and Sara PALACIOS1
1CEPROCOR-ACC, Alvarez de Arenales 230, Barrio Juniors, (X5004AAP) C<Oh><0225><Wa>ordoba, Argentina 2Entomological Research Center, National University of Cordoba, Av. Velez Sarsfield 299 (5000) C<Oh><0225><Wa>ordoba, Argentina
Received January 7, 2002; Accepted April 12, 2002
Systematic fractionation of a fruit extract from Argentine Melia azedarach L., which was monitored by an insect antifeedant bioassay, led to the isolation of meliartenin, a limonoid antifeedant, which existed as a mixture of two interchangeable isomers. At 4 μg/cm2 and 1 μg/cm2, the isomeric mixture was as active as azadirachtin in strongly inhibiting the larval feeding of Epilachna paenulata Germ. (Coleoptera: Coccinellidae) and the polyphagous pest, Spodoptera eridania (Lepidoptera: Noctuidae), respectively.
Key words: Melia azedarach; Meliaceae; meliartenin; insect antifeedant; limonoid
-19-
Note
Sex Differences in Kidney Mineral Concentrations and Urinary Albumin Excretion
in Rats Given High-phosphorus Feed
Hiroshi MATSUZAKI,1 Shin-ichi KATSUMATA,2 Ritsuko MASUYAMA,2 Mariko UEHARA,2 Kahoru NAKAMURA,1 and Kazuharu SUZUKI2
1Department of Nutrition, Junior College of Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan 2Department of Nutritional Science, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan
Received January 9, 2002; Accepted April 6, 2002
We examined sex differences in kidney mineral concentrations and urinary albumin excretion in rats given feed containing various phosphorus (P) levels. With feed that was 0.6％, 0.9％, 1.2％, and 1.5％P, kidney calcium and P concentrations were higher in female rats than in male rats. With 1.2％ or 1.5％P, urinary albumin excretion was higher in the female rats. The sex of the animal affected the kidney mineral concentrations and urinary albumin excretion in rats with a high P intake.
Key words: sex difference; kidney mineral concentration; urinary albumin excretion; high phosphorus intake; rats
-20-
Note
Alternative Splicing Products of the Gene for a Human Nuclear Actin-related
Protein, hArpNβ/Baf53, that Encode a Protein Isoform, hArpNβS, in the Cytoplasm
Eri OHFUCHI, Katsuhiko NISHIMORI, and Masahiko HARATA
Laboratory of Molecular Biology, Department of Molecular and Cell Biology, Graduate School of Agricultural Science, Tohoku University, Tsutsumidori-Amamiyamachi 1-1, Aoba-ku, Sendai 981-8555, Japan
Received January 11, 2002; Accepted April 5, 2002
A human nuclear actin-related protein, hArpNβ/Baf53, is a component of chromatin remodeling and histone acetyltransferase complexes. We identified two alternative splicing products of the gene for hArpNβ/Baf53. They encoded a protein isoform, hArpNβS; and its fusion product with green fluorescent protein was to be found in the cytoplasm, not the nucleus. The isoforms may contribute to functional regulation of these complexes.
Key words: actin-related protein; alternative splicing; subcellular location; chromatin remodeling
-21-
Note
Iron Absorption in Rats Increased by Yeast Glucan
Ta Thi Tuyet MAI, Kaori IGARASHI, Rieko HIRUNUMA, Satoko TAKASAKI, Masaaki YASUE, Shuichi ENOMOTO, Shuichi KIMURA, and Nguyen Van CHUYEN
Department of Food and Nutrition, Japan Women′s University Graduate School of Human Life Sciences, Showa Women′s University Radioisotope Technology Division, Cyclotron Center, RIKEN Asahi Breweries, Ltd.
Received January 16, 2002; Accepted April 9, 2002
The effects of brewer′s yeast cell walls and two of its components, glucan and mannan, on the absorption of 59Fe by anemic rats were investigated. After administration of the label, the percentage of 59Fe taken up into the blood of group given glucan was generally similar to that of a group given yeast cell walls, both values were higher than in controls. The incorporation of 59Fe into the small intestines was higher in the group given glucan than in the controls or a group given a glucan---mannan mixture. Glucan is the main substance in yeast cell walls that increases iron absorption.
Key words: brewer′s yeast; yeast; iron absorption; glucan; mannan
-22-
Note
Shogaols from Zingiber officinale as Promising Antifouling Agents
Hideo ETOH, Takeyoshi KONDOH, Rikoh NODA, Inder Pal SINGH, Yohko SEKIWA, Kohjiro MORIMITSU, and Kikue KUBOTA
Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan Department of Nutrition and Food Science, Ochanomizu University, 2-1-1 Ohtsuka, Bunkyo-ku, Tokyo 112-8610, Japan
Received January 16, 2002; Accepted April 3, 2002
We isolated the highly potent attachment-inhibitors (three times more active than standard CuSO4 in the blue mussel assay), trans-6-, 8-, and 10-shogaols, from a hexane extract of the roots of ginger, Zingiber officinale Roscoe. Trans-8-shogaol showed the highest antifouling activity comparable with that of tributyltin fluoride (TBTF), which is recognized as one of the most effective antifouling agents, in the conventional submerged assay.
Key words: Zingiber officinale Roscoe; shogaol; attachment-inhibiting activity; antifouling; Mytilus edulis galloprovincialis
-23-
Note
Composition of Seed Storage Proteins Changed by Glutathione Treatment of Soybeans
Motoko AWAZUHARA,1,3 Hoyeun KIM,1,2 Hiroaki HAYASHI,1 Mitsuo CHINO,1,4 Sang-Gu KIM,3 and Toru FUJIWARA1,
1Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan 2School of Biological Sciences, Seoul National University, Seoul 151-742, Korea
Received January 21, 2002; Accepted April 10, 2002
The application of glutathione to immature soybean cotyledons reduced the accumulation of the β subunit of β-conglycinin, and increased the accumulation of most glycinins. Both reduced and oxidized forms of glutathione had these effects. The application of an inhibitor of glutathione synthesis, buthionine sulfoximine, increased accumulation of β subunit. These results suggest that glutathione is important in affecting the composition of seed storage proteins.
Key words: glutathione; β subunit of β-conglycinin; seed storage proteins; buthionine sulfoximine; cotyledon culture
-24-
Note
Improving Effect of Dietary Taurine Supplementation on the Oxidative Stress
and Lipid Levels in the Plasma, Liver and Aorta of Rabbits Fed on a High-Cholesterol
Diet
Jale BALKAN,1, <Oh><0196><Wa>Oznur KANBA<Oh><0188><Wa>GLI,1 Aydan HAT<Oh>_<Wa>IPO<Oh><0188><Wa>GLU,1 Mutlu K<Oh><0196><Wa>U<Oh><0199><0196><Wa>UK,2 U<Oh><0178><Wa>gur <Oh><0199><Wa>EV<Oh>_<Wa>IKBAflS,3 Gul<Oh><0231><Wa>in AYKA<Oh><0199><Wa>-TOKER,1 and M<Oh><0228><Wa>ujdat UYSAL1
1Department of Biochemistry, Istanbul Medical Faculty, University of Istanbul, <Oh><0199><Wa>apa 34390, Istanbul, Turkey 2The Experimental and Medical Research Center, University of <Oh>_<Wa>Istanbul 3Department of Pathology, Istanbul Medical Faculty, University of Istanbul
Received January 21, 2002; Accepted March 29, 2002
The effect of a high-cholesterol diet with or without taurine on lipids and oxidative stress in the plasma, liver and aorta of rabbits was investigated. The animals were maintained on a basal diet (control), a high-cholesterol diet (HC, 1％ w/w), or a high- cholesterol diet supplemented with taurine (HCHT, 2.5％ w/w) for two months. Taurine has an ameliorating effect on atherosclerosis together with a decreasing effect on the cholesterol and triglyceride levels in rabbits fed on an HC diet. The HCHT diet caused a significant decrease in the malondialdehyde (MDA) and diene conjugate (DC) levels in the plasma, liver and aorta of rabbits as compared to the HC group. This treatment did not alter the antioxidant system in the liver of rabbits in the HC group. Our findings indicate that taurine ameliorated oxidative stress and cholesterol accumulation in the aorta of rabbits fed on the HC diet and that this effect may be related to its antioxidative potential as well as its reducing effect on serum lipids.
Key words: taurine; hypercholesterolemia; lipid peroxidation; antioxidant; rabbit
-25-
Note
Effects of Lentinus edodes on Fatty Acid and Molecular Species Profiles of Phosphatidylcholine
in Rats Fed Different Levels of Corn Oil
Yasuhiko SHIMADA, Tatsuya MORITA, and Kimio SUGIYAMA
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan
Received January 22, 2002; Accepted March 29, 2002
Shiitake mushrooms (Lentinus edodes) are a hypocholesterolemic and affect phospholipid and fatty acid metabolism in rats. In this study, the effects of 2％ shiitake in the diet on fatty acid and molecular species profiles of liver microsomal and plasma phosphatidylcholine (PC) were investigated in rats fed diets containing different levels (1--20％) of corn oil, a linoleic-acid-rich fat. The proportion of 18:2n-6 in PC increased depending on the parcent corn oil, and L. edodes further increased the proportion at all corn oil levels. The proportion of 20:4n-6 was lower in rats fed L. edodes than in rats fed control diets irrespective of the parcent corn oil. L. edodes selectively increased the proportion of 16:0--18:2 molecular species and decreased the proportion of 18:0--20:4 molecular species in PC. These results indicate that the effects of L. edodes on fatty acid and molecular species profiles of PC are stronger than that of the dietary corn oil level.
Key words: Lentinus edodes; eritadenine; phosphatidylcholine; corn oil; linoleic acid metabolism
-26-
Note
Synthetic Studies of Proanthocyanidins. Highly Stereoselective Synthesis of
the Catechin Dimer, Procyanidin-B3
Akiko SAITO,1 Noriyuki NAKAJIMA,2 Akira TANAKA,3 and Makoto UBUKATA2,
1Japan Science and Technology Corporation; Domestic Research Fellow 2Biotechnology Research Center, Toyama Prefectural University 3College of Technology, Toyama Prefectural University Kosugi, Toyama 939-0398, Japan
Received January 24, 2002; Accepted April 16, 2002
A stereoselective synthesis of benzylated procyanidin-B3, a condensed catechin dimer, is described. Condensation of 5,7,3′,4′-tetrabenzylcatechin with (2R,3S,4S)-5,7,3′,4′-tetrabenzyloxy-3-acetoxy-4-methoxyflavan as an electrophile in the presence of TiCl4 led to octabenzylated procyanidin-B3 stereoselectively.
Key words: polyphenol; proanthocyanidin; stereoselective synthesis; condensed tannins
-27-
Note
Tocotrienol Levels in Various Tissues of Sprague-Dawley Rats after Intragastric
Administration of Tocotrienols
Masaaki OKABE, Mariko OJI, Ikuo IKEDA, Hirofumi TACHIBANA, and Koji YAMADA
Division of Applied Biological Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, Fukuoka 812-8531, Japan
Received February 6, 2002; Accepted April 10, 2002
A tocotrienol (T3) mixture was intragastricaly administered to Sprague-Dawley rats, and the T3 levels in various tissues were measured 0, 4, 8 and 24 hr after the administration. In blood clots, brain, thymus, testes, vice-testes and muscles, T3 homologues were not detected at all. In epididymal adipose, renal adipose, subcutaneous adipose and brown adipose tissues and in the heart, the T3 levels were maintained or increased for 24 hr after the administration. In the serum, liver, mesenteric lymph node, spleen and lungs, the T3 levels were highest 8 hr after the T3 administration. These results suggest that the distribution and metabolism of T3 in the rat vary considerably among different tissues.
Key words: tocotrienol; tocopherol
-28-
Note
Lipase-Catalyzed Kinetic Resolution of (±)-cis-Flavan-4-ol and Its Acetate:
Synthesis of Chiral 3-Hydroxyflavanones
Tamotsu TODOROKI,1 Akiko SAITO,2 and Akira TANAKA1,
1College of Technology, Toyama Prefectural University, Kosugi, Toyama 939-0398, Japan 2Japan Science and Technology Corporation, Domestic Research Fellow, Kosugi, Toyama 939-0398, Japan
Received February 6, 2002; Accepted March 25, 2002
Lipase-catalyzed kinetic resolution of (±)-cis-flavan-4-ol and its acetate led to enantiomerically enriched flavan-4-ol and its acetate. These chiral compounds were converted to (2R, 3R)- and (2S, 3S)-3-hydroxyflavanones.
Key words: kinetic resolution; lipase; 3-hydroxyflavanone
-29-
Note
Purification and Properties of a Carbonyl Reductase Useful for Production of
Ethyl (S)-4-Chloro-3-hydroxybutanoate from Kluyveromyces lactis
Hiroaki YAMAMOTO, Norihiro KIMOTO, Akinobu MATSUYAMA, and Yoshinori KOBAYASHI
Life Science Development Center, CPI Company, Daicel Chemical Industries, Ltd., 27 Miyukigaoka, Tsukuba, Ibaraki 305-0841, Japan
Received February 13, 2002; Accepted March 27, 2002
A novel carbonyl reductase (KLCR1) that reduced ethyl 4-chloroacetoacetate (ECAA) to synthesize ethyl (S)--4-chloro-3-hydroxybutanoate ((S)-ECHB) was purified from Kluyveromyces lactis. KLCR1 catalyzed the NADPH-dependent reduction of ECAA enantioselectively but not the oxidation of (S)--ECHB. From partial amino acid sequences, KLCR1 was suggested to be an α subunit of fatty acid synthase (FAS) but did not have FAS activity.
Key words: carbonyl reductase; Kluyveromyces lactis; ethyl (S)-4-chloro-3-hydroxybutanoate; fatty acid synthase
-30-
Note
Determination of the Stereochemistry of (－)-Koninginin A by an X-ray Analysis
of Its Synthetic Sample
Kenji MORI,1,‡ Masahiko BANDO,2 and Kaoru ABE3
1Insect Pheromone and Traps Division, Fuji Flavor Co. Ltd., Midorigaoka 3-5-8, Hamura-City, Tokyo 205-8503, Japan 2Organic Chemistry Research Institute, Otsuka Pharmaceutical Co. Ltd., Kawauchi, Tokushima 771-0192, Japan 3Tokushima Second Factory, Otsuka Pharmaceutical Co. Ltd., Kawauchi, Tokushima 771-0192, Japan
Received March 18, 2002; Accepted April 22, 2002
The absolute configuration of (－)-koninginin A ｛10-hexyl-11,12-dioxatricyclo[7.2.1.0〈1,6〉]dodecane-
2,5-diol (1)｝, the antibiotic metabolite of Trichoderma koningii and T. harzianum, was determined as 1S, 2R, 5S, 6S, 9S, 10S by an X-ray crystallographic analysis of its synthetic sample coupled with the established stereochemical outcome of Sharpless asymmetric dihydroxylation used as the key reaction to prepare intermediate 4.
Key words: antibiotics; koninginin A; Trichoderma koningii; Trichoderma harzianum; X-ray analysis<EL>