(Vol.68 No.1 2004)
Review
Molecular Dissection of the Selenomonas ruminantium Cell Envelope and Lysine
Decarboxylase Involved in the Biosynthesis of a Polyamine Covalently Linked
to the Cell Wall Peptidoglycan Layer
Yumiko Takatsuka and Yoshiyuki Kamio† p.1
Distribution of Prx-linked Hydroperoxide Reductase Activity among
Microorganisms
Kouji Takeda,1 Yoshitaka Nishiyama,2 Koji Yoda,3 Toshihiro Watanabe,4 Kaori
Nimura-Matsune,1 Kiyoshi Mura,2 Chiyoko Tokue,2 Tetzuya Katoh,1 Shinji Kawasaki,1
and Youichi Niimura1,† p.20
Rare Bacterium of New Genus Isolated with Prolonged Enrichment
Culture
Akiko Hashizume,1 Ryosuke Fudou,2 Yasuko Jojima,2 Ryohsuke Nakai,1 Akira Hiraishi,3
Akira Tabuchi,1 Kikuo Sen,1 and Hiroshiro Shibai1,† p.28
Antioxidative Effects of Glycosyl-ascorbic Acids Synthesized by
Maltogenic Amylase to Reduce Lipid Oxidation and Volatiles Production in Cooked
Chicken Meat
Soo-Bok Lee,1,† Ki-chang Nam,2 Sung-Joon Lee,3 Jong-Ho Lee,1 Kuniyo Inouye,4
and Kwan-Hwa Park5 p.36
Effects of Excess Nicotinamide Administration on the Urinary Excretion
of Nicotinamide N-Oxide and Nicotinuric Acid by Rats
Tsutomu Fukuwatari,† Hideko Wada, Ryuzo Sasaki, and Katsumi Shibata p.44
Simultaneous Determination of Isoflavones and Bisphenol A in Rat
Serum by High-performance Liquid Chromatography Coupled with Coulometric Array
Detection
Shin Yasuda,1,† Po-Sheng Wu,1 Emi Hattori,1 Hirofumi Tachibana,1 and Koji
Yamada1,2 p.51
Isolation of Tryptophan as an Inhibitor of Ovalbumin Permeation
and Analysis of Its Suppressive Effect on Oral Sensitization
Jun Watanabe,1,2 Kyoko Fukumoto,1 Eri Fukushi,1 Kei Sonoyama,1,† and Jun
Kawabata1 p.59
Encapsulation of Shiitake (Lenthinus Edodes) Flavors by Spray Drying
Hirokazu Shiga,1 Hidefumi Yoshii,1,† Hisashi Ohe,1 Masahumi Yasuda,1 Takeshi
Furuta,1 Hiroshige Kuwahara,2 Masaaki Ohkawara,3 and Pekka Linko4 p.66
Comparison of the Changes in Lipid Metabolism between Hepatoma-bearing
and Lipopolysaccharide-treated Rats
Masashi Kawasaki,† Kazumi Yagasaki, Yutaka Miura, and Ryuhei Funabiki
p.72
Catalytic Reaction of Basidiomycete Lentinula edodes Cytochrome
P450, Le.CYP1 Enzyme Produced in Yeast
Ryoko Akiyama, Susumu Kajiwara, and Kazuo Shishido† p.79
Triterpene Acids from the Leaves of Perilla frutescens and Their
Anti-inflammatory and Antitumor-promoting Effects
Norihiro Banno,1 Toshihiro Akihisa,2,† Harukuni Tokuda,3 Ken Yasukawa,4
Hiroshi Higashihara,3 Motohiko Ukiya,2 Kenji Watanabe,2 Yumiko Kimura,4 Jun-ichi
Hasegawa,1 and Hoyoku Nishino3 p.85
Decomposition Kinetics of Maltose in Subcritical Water
Shabnam Haghighat Khajavi, Yukitaka Kimura, Toshinobu Oomori, Ryuichi Matsuno,
and Shuji Adachi† p.91
High Level Expression of Thermostable Lipase from Geobacillus
sp. Strain T1
Thean Chor Leow, Raja Noor Zaliha Raja Abdul Rahman,† Mahiran Basri, and
Abu Bakar Salleh p.96
Tetrapetalone A, a Novel Lipoxygense Inhibitor from Streptomyces
sp.
Toshikazu Komoda,1 Kayo Yoshida,1 Naoki Abe,1 Yasumasa Sugiyama,1 Misako Imachi,2
Hiroshi Hirota,3,4 Hiroyuki Koshino,5 and Akira Hirota1,† p.104
Proteomic Identification of α-Amylase Isoforms Encoded by
RAmy3B/3C from Germinating Rice Seeds
Yohei Nanjo,1 Satoru Asatsuma,1 Kimiko Itoh,1,2 Hidetaka Hori,1 and Toshiaki
Mitsui1,2,3,† p.112
A Hydroxyl Group of Flavonoids Affects Oral Anti-inflammatory
Activity and Inhibition of Systemic Tumor Necrosis Factor-α Production
Hiroshi Ueda,† Chikako Yamazaki, and Masatoshi Yamazaki p.119
Anisodamine Causes the Changes of Structure and Function in the
Transmembrane Domain of the Ca2+-ATPase from Sarcoplasmic Reticulum
Yu-Hong Pang and Jian-Wen Chen† p.126
Bacillus subtilis ypgA Gene Is fni, a Nonessential Gene Encoding
Type 2 Isopentenyl Diphosphate Isomerase
Motoki Takagi, Kazuhide Kaneda,* and Tomohisa Kuzuyama† p.132
Royal Jelly Inhibits the Production of Proinflammatory Cytokines
by Activated Macrophages
Keizo Kohno,1,2,† Iwao Okamoto,1 Osamu Sano,1 Norie Arai,1 Kanso Iwaki,1
Masao Ikeda,1 and Masashi Kurimoto1 p.138
Cloning of a Gene Cluster Responsible for the Biosynthesis of
Diterpene Aphidicolin, a Specific Inhibitor of DNA Polymerase α
Tomonobu Toyomasu,1 Kentaro Nakaminami,1 Hiroaki Toshima,2 Takashi Mie,3 Kenji
Watanabe,3 Hiroyuki Ito,3 Hirokazu Matsui,3 Wataru Mitsuhashi,1 Takeshi Sassa,1
and Hideaki Oikawa4,† p.146
Chiral Discrimination of Branched-chain Fatty Acids by Reversed-phase
HPLC after Labeling with a Chiral Fluorescent Conversion Reagent
Kazuaki Akasaka and Hiroshi Ohrui† p.153
The Primary Structure of a Novel Goose-type Lysozyme from Rhea
Egg White*
Jureerut Pooart, Takao Torikata, and Tomohiro Araki† p.159
Metofluthrin: A Potent New Synthetic Pyrethroid with High Vapor
Activity against Mosquitoes
Kazuya Ujihara,† Tatsuya Mori, Tomonori Iwasaki, Masayo Sugano, Yoshinori
Shono, and Noritada Matsuo p.170
Frost-susceptible Protein in Plasma Membranes in Tubers of Helianthus
tuberosus L.
Keita Arakawa,† Mitsuru Hanazaki, and Shizuo Yoshida p.175
Synthesis and Antioxidant Activity of Oxygenated Furofuran Lignans
Satoshi Yamauchi,1,† Tomoko Ina,1 Takuya Kirikihira,2 and Toshiya Masuda2
p.183
Preparation of Selenium-enriched Sprouts and Identification of
Their Selenium Species by High-performance Liquid Chromatography-Inductively
Coupled Plasma Mass Spectrometry
Satoru Sugihara, Mariko Kondo, Yuko Chihara, Mami Yuji, Hiroyuki Hattori, and
Munehiro Yoshida† p.193
Tomato Paste Fraction Inhibiting the Formation of Advanced Glycation
End-products
Tadashi Kiho,1,† Shigeyuki Usui,2 Kazuyuki Hirano,2 Koichi Aizawa,3 and
Takahiro Inakuma3 p.200
Self-cloning Yeast Strains Containing Novel FAS2 Mutations Produce
a Higher Amount of Ethyl Caproate in Japanese Sake
Kazuo Aritomi,1 Isao Hirosawa,2 Hisashi Hoshida,2 Mikio Shiigi,1 Yoshinori Nishizawa,2
Susumu Kashiwagi,1 and Rinji Akada2,† p.206
Note
Effects of Several Variable Factors on the Isotope Ratio by HRGC-MS
Masayoshi Sawamura,1,† Atsushi Satake,1,2 Takao Ueno,1 Akitoshi Une,1
and Hiroyuki Ukeda1 p.215
Note
σmarY1, the LTR of the gypsy-Type Retroelement marY1 from the Basidiomycete
Tricholoma matsutake, Allows Multicopy DNA Integration in Lentinula edodes*
Hitoshi Murata† and Yasumasa Miyazaki p.218
Note
Detection of Hydrolytic Activity of Trypsin with a Fluorescence-chymotryptic
Peptide on a TLC Plate
Tetsuya Uchikoba,1,† Shigeko Fukumoto,1 Takao Itakura,2 Michiko Okubo,3
Kazuhiko Tomokiyo,4 Kazunari Arima,5 and Hiroo Yonezawa5 p.222
Note
SMXA-5 Mouse as a Diabetic Model Susceptible to Feeding a High-fat Diet
Misato Kobayashi,1 Fusayo Io,1 Takahiro Kawai,1 Masahiko Nishimura,2 Tamio Ohno,2
and Fumihiko Horio1,† p.226
Note
Distribution of Minerals in Quinoa (Chenopodium quinoa Willd.) Seeds
Yotaro Konishi,1,† Shigeru Hirano,1,2 Hideki Tsuboi,3 and Masao Wada3
p.231
Note
High-level Production of Hyperthermophilic Cellulase in the Bacillus brevis
Expression and Secretion System
Yasuhiro Kashima1 and Shigezo Udaka2,† p.235
Note
Suppressive Effects of Selected Antioxidants on the Activated Leukocytes-induced
Mutagenesis in the Co-culture Assay Systems
Ha Won Kim,1 Akira Murakami,1 Marshall V. Williams,2 and Hajime Ohigashi1,†
p.238
Note
High Phosphorus Diet Changes Phosphorus Metabolism Regardless of PTH Action
in Rats
Shin-ichi Katsumata,1 Ritsuko Masuyama,1 Moyuru Koshihara,1 Hiroshi Matsuzaki,2
Mariko Uehara,1 and Kazuharu Suzuki1,† p.243
Note
Protective Effect of Blue-M1 against Oxidative Stress on COS-1 Cells
Teruyuki Usui,1 Satomi Shizuuchi,1 Hirohito Watanabe,2 and Fumitaka Hayase1,†
p.247
Note
Maturation of Fermented Rice-koji Miso Can Be Monitored by an Increase in Fatty
Acid Ethyl Ester
Shigeo Yamabe,1,† Kentaro Kaneko,2 Hiroko Inoue,3 and Toshichika Takita3
p.250
Note
Introduction of DPR, an Enterostatin Fragment Peptide, into Soybean β -Conglycinin
α ’ Subunit by Site-directed Mutagenesis
Yasuyuki Takenaka,1,* Naomi Doyama,1 Nobuyuki Maruyama,2 Shigeru Utsumi,2 and
Masaaki Yoshikawa1,† p.253
Note
Penipratynolene, a Novel Nematicide from Penicillium bilaiae Chalabuda
Satoshi Nakahara,1 Miyako Kusano,2 Shozo Fujioka,3 Atsumi Shimada,4 and Yasuo
Kimura1,† p.257
Note
A New Prenylated Flavonoid from Propolis Collected in Okinawa, Japan
Shigenori Kumazawa,1,† Hitomi Goto,1 Tomoko Hamasaka,1 Syuichi Fukumoto,2
Takunori Fujimoto,3 and Tsutomu Nakayama1 p.260
Communication
Mac1 Positive Cells Are Required for Enhancement of Splenocytes Proliferation
Caused by Bisphenol A
Masao Goto,† Hiroshi Ono, Yuko Takano-Ishikawa, Hiroshi Shinmoto, and
Mitsuru Yoshida p.263
Communication
Truncated Sla1 Induces Haploid Meiosis through the Pat1-Mei2 System in Fission
Yeast
Kaori Tanabe, Katsunori Tanaka, Hideyuki Matsuda, and Makoto Kawamukai†
p.266
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Review
Molecular Dissection of the Selenomonas ruminantium Cell Envelope and Lysine
Decarboxylase Involved in the Biosynthesis of a Polyamine Covalently Linked
to the Cell Wall Peptidoglycan Layer
Yumiko Takatsuka and Yoshiyuki Kamio†
Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumi-dori Amamiya-machi, Aoba-ku, Sendai 981-8555, Japan
The wild type of Selenomonas ruminantium subsp. lactilytica, which is a strictly anaerobic, Gram-negative bacterium isolated from sheep rumen, requires one of the normal saturated volatile fatty acids with 3 to 10 carbon atoms for its growth in a glucose medium; however, no such obligate requirement of fatty acid is observed when the cells are grown in a lactate medium. This bacterium is characterized by a unique structure of the cell envelope and a novel lysine decarboxylase and its regulatory protein. In the first part of this article, we will refer to the chemical structure of phospholipid and lipopolysaccharide in the cell membranes of this bacterium compared with that from the general Gram-negative bacteria for understanding their biological functions. S. ruminantium has neither free nor bound forms of Braun lipoprotein which plays an important role of the maintenance of the structural integrity of the cell surface in general Gram-negative bacteria. However, S. ruminantium has cadaverine, which links covalently to the peptidoglycan as a pivotal constituent for the cell division. In the second part of this article, we will refer to the chemical structure of the cadaverine-containing peptidoglycan, its biosynthesis, and the biological function. In the third part of this article, we will depict the molecular cloning of the genes encoding S. ruminanitum lysine decarboxylase (LDC) and its regulatory protein of 22-kDa (22-kDa protein; P22) which has similar characteristics to that of antizyme of ornithine decarboxylase in eukaryotic cells, and the molecular dissection of these proteins for understanding the regulation of cadaverine biosynthesis. Finally, we will illustrate a proposed structure of the cell envelope, a processes of biosynthesis of the cadaverine-containing peptidoglycan layer, and the LDC degradation mechanism in S. ruminantium, on the basis of the analyses of the cell envelope components, the results from the in vitro experiments on the biosynthesis of the peptidoglycan layer, and the current status of the knowledge on LDC and P22 in this organism.
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Distribution of Prx-linked Hydroperoxide Reductase Activity among Microorganisms
Kouji Takeda,1 Yoshitaka Nishiyama,2 Koji Yoda,3 Toshihiro Watanabe,4 Kaori Nimura-Matsune,1 Kiyoshi Mura,2 Chiyoko Tokue,2 Tetzuya Katoh,1 Shinji Kawasaki,1 and Youichi Niimura1,†
1Department of Bioscience, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan
2Department of Nutritional Science, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan
3Department of Biotechnology, The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan
4Department of Food Science and Technology, Tokyo University of Agriculture, Abashiri-shi, Hokkaido 099-2493, Japan
Peroxiredoxin (Prx) constitutes a large family of enzymes found in microorganisms, animals, and plants, but the detection of the activities of Prx-linked hydroperoxide reductases (peroxiredoxin reductases) in cell extracts, and the purification based on peroxide reductase activity, have only been done in bacteria and Trypanosomatidae. A peroxiredoxin reductase (NADH oxidase) from a bacterium, Amphibacillus, displayed only poor activities in the presence of purified Prx from Saccharomyces or Synechocystis, while it is highly active in the presence of bacterial Prx. These results suggested that an enzyme system different from that in bacteria might exist for the reduction of Prx in yeast and cyanobacteria.
Prx-linked hydroperoxide reductase activities were detected in cell extracts of Saccharomyces, Synechocystis, and Chlorella, and the enzyme activities of Saccharomyces and Chlorella were induced under vigorously aerated culture conditions and intensive light exposure conditions, respectively. Partial purification of Prx-linked peroxidase from the induced yeast cells indicated that the Prx-linked peroxidase system consists of two protein components, namely, thioredoxin and thioredoxin reductase. This finding is consistent with the previous report on its purification based on its protein protection activity against oxidation [Chae et al., J. Biol. Chem., 269, 27670-27678 (1994)]. In this study we have confirmed that Prx-linked peroxidase activity are widely distributed, not only in bacteria species and Trypanosomatidae, but also in yeast and photosynthetic microorganisms, and showed reconstitution of the activity from partially purified interspecies components.
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Rare Bacterium of New Genus Isolated with Prolonged Enrichment Culture
Akiko Hashizume,1 Ryosuke Fudou,2 Yasuko Jojima,2 Ryohsuke Nakai,1 Akira Hiraishi,3 Akira Tabuchi,1 Kikuo Sen,1 and Hiroshiro Shibai1,†
1Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, 8304, Minamiminowa, Nagano 399-4511, Japan
2Institute of Life Science, Ajinomoto Co., Inc., Suzukicho 1-1, Kawasaki-shi, Kanagawa 210-8681, Japan
3Department of Ecological Engineering Toyohashi University of Technology, Tenpakucho Hibarigaoka 1-1, Toyohashi-shi, Aichi 441-8580, Japan
Dynamic change in microbial flora was monitored with an oxygen electrode. The 1st phase microorganisms, which first grew well in LB medium, were followed by the 2nd phase microorganisms, which supposedly assimilated microbial cells of the 1st phase and their metabolites. In a similar way, a change in microbial flora was observed from the 1st phase to the 4th phase in 84 hr. Based on this observation, prolonged enrichment culture was done for as long as two months to increase the ratio of existence of rare microorganisms.
>From these culture liquids, four slow-growing bacteria (provisionally named Shinshu-ah1, -ah2, -ah3, and -ah4), which formed scarcely visible small colonies, were isolated. Sequence analysis of their 16S rDNA showed that Shinshu-ah1 had 97% homology with Bradyrhizobium japonicum and uncultured alpha proteobacterium clone blaii 16, Shinshu-ah2 91% with Rasbo bacterium, Alpha proteobacterium 34619, Bradyrhizobium genosp. P, Afipia felis and an unidentified bacterium, Shinshu-ah3 99% with Methylobacterium mesophilicum, and Shinshu-ah4 95% with Agromyces ramosus DSM 43045. Phylogenetic study indicated that Shinshu-ah2 had a possibility to form a new family, Shinshu-ah1 a new genus, and Shinshu-ah4 a new species.
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Antioxidative Effects of Glycosyl-ascorbic Acids Synthesized by Maltogenic Amylase
to Reduce Lipid Oxidation and Volatiles Production in Cooked Chicken Meat
Soo-Bok Lee,1,† Ki-chang Nam,2 Sung-Joon Lee,3 Jong-Ho Lee,1 Kuniyo Inouye,4 and Kwan-Hwa Park5
1Research Institute of Food and Nutritional Sciences and Department of Food and Nutrition, Yonsei University, Seoul 120-749, Korea
2Department of Animal Science, Iowa State University, Ames, Iowa 50011-3150, USA
3Palo Alto Medical Foundation Research Institute, Stanford University, Palo Alto, California, USA
4Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
5Center for Agricultural Bio-Materials and Department of Food Science and Technology, School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea
Glycosylated ascorbic acids were synthesized by using the transglycosylation activity of Bacillus stearothermophilus maltogenic amylase with maltotriose to show effective antioxidative activity with enhanced oxidative stability. The modified ascorbic acids comprised mono- and di-glycosyl transfer products with an α-(1,6)-glycosidic linkage. The antioxidative effects of the glycosyl derivatives of ascorbic acid on the lipid oxidation of cooked chicken breast meat patties were compared, and the synergistic effect when combined with α-tocopherol was determined in terms of thiobarbituric acid-reactive substances (TBARS) and volatiles production during storage. The results indicate that the glycosylated ascorbic acids had very effective antioxidative activity in preventing lipid oxidation, and were better in their synergistic effect in comparison to authentic ascorbic acid, with maltosyl-ascorbic acid being the most effective. Volatiles production was highly correlated with the TBARS values in the lipid oxidation of cooked meat. The antioxidative effect preventing the production of volatiles was particularly strong on pentanal, fairly strong on propanal and butanal, and not at all on ethanal. Propanal, pentanal, and the total volatiles thus provided a good representation of the lipid oxidation status of cooked chicken meat.
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Effects of Excess Nicotinamide Administration on the Urinary Excretion of Nicotinamide
N-Oxide and Nicotinuric Acid by Rats
Tsutomu Fukuwatari,† Hideko Wada, Ryuzo Sasaki, and Katsumi Shibata
Laboratory of Food Science and Nutrition, Department of Life Style Studies, School of Human Cultures, The University of Shiga Prefecture, Hikone, Shiga 522-8533, Japan
We investigated a useful chemical index for an excessive nicotinamide intake and how this excessive nicotinamide intake affects the tryptophan-nicotinamide metabolism in rats. Weaning rats were fed on a tryptophan-limited and nicotinic acid-free diet containing no, 0.003%, 0.1%, 0.2%, or 0.3% nicotinamide for 21days. Urine samples were collected on the last day and analyzed the intermediates and metabolites on the tryptophan-nicotinamide pathway. Nicotinamide N-oxide, nicotinic acid and nicotinuric acid, metabolites of nicotinamide, were detected when nicotinamide at more than 0.1% had been taken. An intake of nicotinamide of more than 0.1% increased the urinary excretion of quinolinic acid, an intermediate on the pathway. Nicotinamide N-oxide and nicotinuric acid increased with increasing dietary concentration of nicotinamide. These results show that the measurements of nicotinamide N-oxide and nicotinuric acid in urine would be useful indices for an excessive nicotinamide intake.
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Simultaneous Determination of Isoflavones and Bisphenol A in Rat Serum by High-performance
Liquid Chromatography Coupled with Coulometric Array Detection
Shin Yasuda,1,† Po-Sheng Wu,1 Emi Hattori,1 Hirofumi Tachibana,1 and Koji Yamada1,2
1Laboratory of Food Chemistry, Division of Applied Biological Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan
2CREST, Japan Science and Technology, Japan
A method for simultaneous detection and quantification is presented to determine the presence of isoflavones and bisphenol A in a biological sample. A coulometric array detector was used with reversed-phase high-performance liquid chromatography (HPLC). Daidzein (1), glycitein (2), genistein (3) and their glucoside conjugates, daidzin (4), glycitin (5) and genistin (6), were measured as phytochemicals. Also assayed here was equol (7), a metabolite from compound 1, and bisphenol A (8), an industrial chemical that acts as an endocrine disrupter. All chemicals were simultaneously detected by using a 600-mV single detection voltage with high efficacy. A mixture of 1, 3 and 8 was orally administered to rats, and the levels of these three chemicals in the serum were clearly increased after a 4 kU β-glucuronidase treatment. The levels of compounds 1 and 3 in the serum were detected at 1665 and 2040 ng/ml, while 8 was at a low level of 417 ng/ml. Compound 7 in the serum was not detected until after enzymatic hydrolysis (72 ng/ml). These results suggest that this analytical method would be useful for metabolic and pharmacokinetic studies on isoflavones and bisphenol A.
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Isolation of Tryptophan as an Inhibitor of Ovalbumin Permeation and Analysis
of Its Suppressive Effect on Oral Sensitization
Jun Watanabe,1,2 Kyoko Fukumoto,1 Eri Fukushi,1 Kei Sonoyama,1,† and Jun Kawabata1
1Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
2Japan Society for the Promotion of Science, Chiyoda-ku, Tokyo 102-8471, Japan
Tryptophan was isolated from rat feces as an active compound against ovalbumin permeation in an in vitro Caco-2 cell model. Tryptophan dose-dependently inhibited ovalbumin permeation with accompanying increase in transepithelial electric resistance, and its inhibitory activity reached a plateau at 10 mM. Brown Norway rats were sensitized by intragastric administration of ovalbumin together with or without tryptophan. Antibody levels specific to ovalbumin in the sera and proliferative responses of spleen mononuclear cells to ovalbumin were significantly lower in rats administered ovalbumin plus tryptophan than those administered ovalbumin alone. These results suggest that tryptophan suppresses oral sensitization to ovalbumin, probably via suppression of ovalbumin absorption from the intestinal tract.
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Encapsulation of Shiitake (Lenthinus Edodes) Flavors by Spray Drying
Hirokazu Shiga,1 Hidefumi Yoshii,1,† Hisashi Ohe,1 Masahumi Yasuda,1 Takeshi Furuta,1 Hiroshige Kuwahara,2 Masaaki Ohkawara,3 and Pekka Linko4
1Department of Biotechnology, Tottori University, Tottori 680-8552, Japan
2Maruzen Pharmaceuticals Co., Ltd., Onomichi 722-0062, Japan
3Ohkawara Kakouki Co., Ltd., Yokohama 224-0053, Japan
4Department of Chemical Technology, Helsinki University of Technology, P. O. Box 6100, FIN-02015 HUT (Espoo), Finland
Powdery encapsulation of shiitake flavors, extracted from dried shiitake, was investigated by spray drying. Flavor retention increased with an increase in drying air temperature and solid content, and decreased with an increase in dextrose equivalents of maltodextrin. A heat-treatment of the extract liquid made the lenthionine concentration increase, but did not influence the concentrations of the other flavors. The formation of lenthionine with heat-treatment could be described by the consecutive unimolecular-type first order reaction. Lenthionine content in a spray-dried powder prepared with the heated extracted liquid significantly increased. α-Cyclodextrin was the most suitable encapsulant of α-, β-, and γ-cyclodextrins to prepare the spray-dried powder, including lenthionine. The flavor retentions were markedly increased by using of α-cyclodextrin and maltodextrin in combination as an encapsulant.
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Comparison of the Changes in Lipid Metabolism between Hepatoma-bearing and Lipopolysaccharide-treated
Rats
Masashi Kawasaki,† Kazumi Yagasaki, Yutaka Miura, and Ryuhei Funabiki
Department of Applied Biological Science, Tokyo Noko University, Fuchu, Tokyo 183-8509, Japan
To elucidate the mechanism for hyperlipidemia in the hepatoma-bearing state, changes in some parameters related to the lipid metabolism and serum tumor necrosis factor-α (TNF-α) level were examined in Donryu rats that had been subcutaneously implanted with an ascites hepatoma cell line of AH109A. These parameters were also examined in rats that had been given a single injection of lipopolysaccharide (LPS), a model for acute infection with TNF-α secretion into the blood circulation. The serum triglyceride and total cholesterol (Ch) levels were significantly higher in both the hepatoma-implanted and LPS-injected rats than in normal rats. The level of adipose tissue lipoprotein lipase was decreased by hepatoma implantation and LPS injection, while the hormone-sensitive lipase activity was increased by the same treatments. Fatty acid (FA) oxidation and Ch synthesis were also stimulated by both treatments. The serum TNF-α level was noticably elevated by hepatoma implantation and greatly by the LPS injection. This LPS injection increased hepatic FA synthesis. The serum high-density lipoprotein Ch level and hepatic Ch 7α-hydroxylase activity were not changed by the LPS injection. Hepatoma implantation led to hyperlipidemia and elevated the serum TNF-α level, as did the LPS injection.
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Catalytic Reaction of Basidiomycete Lentinula edodes Cytochrome P450, Le.CYP1
Enzyme Produced in Yeast
Ryoko Akiyama, Susumu Kajiwara, and Kazuo Shishido†
Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
The basidiomycete Lentinula edodes (Le.) cytochrome P450, Le.CYP1 was functionally expressed in Saccharomyces cerevisiae. The microsomal fraction containing Le.CYP1 was prepared from the recombinant yeast and the Le.CYP1 was analyzed. The 7-ethoxycoumarin and benzo(a)pyrene were found to be the substrates of Le.CYP1 enzyme. Le.CYP1 converted 7-ethoxycoumarin to 7-hydroxycoumarin.
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Triterpene Acids from the Leaves of Perilla frutescens and Their Anti-inflammatory
and Antitumor-promoting Effects
Norihiro Banno,1 Toshihiro Akihisa,2,† Harukuni Tokuda,3 Ken Yasukawa,4 Hiroshi Higashihara,3 Motohiko Ukiya,2 Kenji Watanabe,2 Yumiko Kimura,4 Jun-ichi Hasegawa,1 and Hoyoku Nishino3
1Ichimaru Pharcos Company Ltd., 318-1 Shinsei, Motosu-gun, Gifu 501-0475, Japan
2College of Science and Technology, Nihon University, 1-8 Kanda Surugadai, Chiyoda-ku, Tokyo 101-8308, Japan
3Department of Biochemistry, Kyoto Prefectural University, Kamigyo-ku, Kyoto 602-0841, Japan
4College of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi-shi, Chiba 274-8555, Japan
Nine triterpene acids, viz., six of the ursane type, ursolic acid (1), corosolic acid (2), 3-epicorosolic acid (3), pomolic acid (4), tormentic acid (5) and hyptadienic acid (6), and three of the oleanane type, oleanolic acid (7), augustic acid (8) and 3-epimaslinic acid (9), among which 1 constituted the most predominant triterpene acid, were isolated and identified from ethanol extracts of the leaves of red perilla [Perilla frutescens (L.) Britton var. acuta Kudo] and green perilla [P. frutescens (L.) Britton var. acuta Kudo forma viridis Makino]. These eight compounds, 1, 2, 4-9, were evaluated for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation (1 μg/ear) in mice. All the compounds tested showed a marked anti-inflammatory effect, with a 50% inhibitory dose (ID50) of 0.09-0.3 mg per ear. In addition, an evaluation against the Epstein-Barr virus early antigen (EBV-EA) activation induced by TPA showed five compounds, 1-3, 5 and 9, with a potent inhibitory effect on EBV-EA induction (91-93% inhibition at 1× 103 mol ratio/TPA). Furthermore, compound 5 exhibited strong antitumor-promoting activity in an in vivo two-stage carcinogenesis test of mouse tumor by using 7,12-dimethylbenz(a)anthracene (DMBA) as an initiator and TPA as a promoter.
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Decomposition Kinetics of Maltose in Subcritical Water
Shabnam Haghighat Khajavi, Yukitaka Kimura, Toshinobu Oomori, Ryuichi Matsuno, and Shuji Adachi†
Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
The decomposition process of maltose in subcritical water was studied using a tubular reactor in the temperature range of 180 to 260°C and at 10 MPa. The formation of glucose and 5-hydroxymethyl-2-furaldehyde during the maltose decomposition was also observed. The decomposition rate of maltose was faster at higher temperatures. The rate was approximated by first-order kinetics during the early stage of the decomposition, but was accelerated and deviated from these kinetics at the later stage. The effluent pH decreased as the residence time in the reactor increased and the decrease of pH affected the maltose decomposition rate and glucose formation. Low pH of a feed solution accelerated maltose decomposition. A good correlation was obtained between the pH of the effluent and the rate constant of the first-order kinetics.
-13-
High Level Expression of Thermostable Lipase from Geobacillus sp. Strain T1
Thean Chor Leow, Raja Noor Zaliha Raja Abdul Rahman,† Mahiran Basri, and Abu Bakar Salleh
Enzyme and Microbial Technology Research Group, Faculty of Science and Environmental Studies, Universiti Putra Malaysia, 43400 UPM Serdang, Malaysia
A thermostable extracellular lipase of Geobacillus sp. strain T1 was cloned in a prokaryotic system. Sequence analysis revealed an open reading frame of 1,251 bp in length which codes for a polypeptide of 416 amino acid residues. The polypeptide was composed of a signal peptide (28 amino acids) and a mature protein of 388 amino acids. Instead of Gly, Ala was substituted as the first residue of the conserved pentapeptide Gly-X-Ser-X-Gly. Successful gene expression was obtained with pBAD, pRSET, pET, and pGEX as under the control of araBAD, T7, T7 lac, and tac promoters, respectively. Among them, pGEX had a specific activity of 30.19 Umg-1 which corresponds to 2927.15 Ug-1 of wet cells after optimization. The recombinant lipase had an optimum temperature and pH of 65°C and pH 9, respectively. It was stable up to 65°C at pH 7 and active over a wide pH range (pH 6-11). This study presents a rapid cloning and overexpression, aimed at improving the enzyme yield for successful industrial application.
-14-
Tetrapetalone A, a Novel Lipoxygense Inhibitor from Streptomyces sp.
Toshikazu Komoda,1 Kayo Yoshida,1 Naoki Abe,1 Yasumasa Sugiyama,1 Misako Imachi,2 Hiroshi Hirota,3,4 Hiroyuki Koshino,5 and Akira Hirota1,†
1Laboratory of Applied Microbiology, School of Food and Nutritional Sciences, University of Shizuoka, Yada 52-1, Shizuoka 422-8526, Japan
2Bruker BioSpin K. K., 3-21-5 Ninomiya, Tsukuba 305-0051, Japan
3Protein Research Group, RIKEN Genomics Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan
4Science of Biological Supramolecular Systems, Yokohama 230-0045, Japan
5Molecular Characterization Team, Advanced D&S Center, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
A simple new assay was designed for lipoxygenase inhibitors. This assay was used to find the novel lipoxygenase inhibitor, tetrapetalone A (1). Tetrapetalone A (1), C26H33NO7, was isolated from Streptomyces sp. USF-4727 strain. Its planar structure was determined by spectroscopic evidence and by methylating with diazomethane to show the presence of a novel tetracyclic skeleton and a β-D-rhodinosyl moiety. The stereochemistry of 1 was investigated by the coupling constant in the 1H-NMR spectrum, NOE correlations, modified Mosher’s method and derivation. We have reported the structural elucidation of 1 in our previous paper. However, further investigation gave another structure for 1, which is described in this paper. Tetrapetalone A showed similar inhibitory activity against soybean lipoxygenase to the two well-known lipoxygenase inhibitors, kojic acid and NDGA, while methylated tetrapetalone A (2) showed little inhibitory activity, even at a concentration of 1 mM.
-15-
Proteomic Identification of α-Amylase Isoforms Encoded by RAmy3B/3C from
Germinating Rice Seeds
Yohei Nanjo,1 Satoru Asatsuma,1 Kimiko Itoh,1,2 Hidetaka Hori,1 and Toshiaki Mitsui1,2,3,†
1Graduate School of Science and Technology, Niigata University, Niigata 950-2181, Japan
2Center for Transdisciplinary Research, Niigata University, Niigata 950-2181, Japan
3Department of Applied Biological Chemistry, Niigata University, Niigata 950-2181, Japan
We isolated and identified 10 α-amylase isoforms by using β-cyclodextrin Sepharose affinity column chromatography and two-dimensional polyacrylamide gel electrophoresis from germinating rice (Oryza sativa L.) seeds. Immunoblots with anti-α-amylase I-1 and II-4 antibodies indicated that 8 isoforms in 10 are distinguishable from α-amylase I-1 and II-4. Peptide mass fingerprinting analysis showed that there exist novel isoforms encoded by RAmy3B and RAmy3C genes. The optimum temperature for enzyme reaction of the RAmy3B and RAmy3C coding isoforms resembled that of α-amylase isoform II-4 (RAmy3D). Furthermore, complex protein polymorphism resulted from a single α-amylase gene was found to occur not only in RAmy3D, but also in RAmy3B.
-16-
A Hydroxyl Group of Flavonoids Affects Oral Anti-inflammatory Activity and Inhibition
of Systemic Tumor Necrosis Factor-α Production
Hiroshi Ueda,† Chikako Yamazaki, and Masatoshi Yamazaki
Department of Medical Life Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko-machi, Kanagawa 199-0195, Japan
We previously reported that oral administration of luteolin can inhibit serum tumor necrosis factor (TNF)-α production and several inflammatory and allergic models. We investigated here the effect of various flavonoids which resemble luteolin in structure. Lipopolysaccharide (LPS)-induced TNF-α production from macrophages was inhibited by treatment with flavone (luteolin, apigenin, and chrysin), flavonol (quercetin and myricetin), flavanonol (taxifolin), and anthocyanidin (cyanidin chloride) in vitro. Most of these, however, did not affect mice when administered orally. Serum TNF-α production was inhibited only by luteolin or apigenin, and only luteolin or quercetin inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema. These results suggest that the structure of luteolin: 3’,4’,5,7-tetrahydroxyflavone, is most suitable for the oral anti-inflammatory activity and that existence or disappearance of a hydroxy group may cause a loss of efficiency.
-17-
Anisodamine Causes the Changes of Structure and Function in the Transmembrane
Domain of the Ca2+-ATPase from Sarcoplasmic Reticulum
Yu-Hong Pang and Jian-Wen Chen†
National Laboratory of Biomacromolecules, Institute of Biophysics, Academia Sinica, Beijing 100101, China
The effects of anisodamine on the Ca2+-ATPsae of sarcoplasmic reticulum (SR) were investigated by using differential scanning calorimetry to measure the ability of anisodamine to denature the transmembrane domain and the cytoplasmic domain. Anisodamine significantly altered the thermotropic phase behaviors of the transmembrane domain of purified Ca2+-ATPase. Specifically, the melting temperature of the transmembrane domain moved toward lower temperatures with the concentrations of anisodamine increasing and the thermotropic phase peak was abolished at 10 mM, indicating that the stabilized structure of the transmembrane domain in the presence of Ca2+ could be destabilized by anisodamine. Decreases of the intrinsic fluorescence and increases of the extrinsic fluorescence of ANS, a fluorescent probe, showed the exposure of tryptophan and hydrophobic region, respectively, suggesting again that anisodamine caused a less compact conformation in the transmembrane domain. A marked inhibition of the Ca2+ uptake activity of SR Ca2+-ATPase was observed when the addition of anisodamine. The drug did not affect the cytoplasmic domain of the enzyme and only slightly decreased the ATPase activity of the enzyme at concentrations up to 10 mM. This was likely due to the destabilized protein transmembrane domain. To sum up, our results revealed that anisodamine interacted specifically with the transmembrane domain of SR Ca2+-ATPase and inhibited the Ca2+ uptake activity of the enzyme.
-18-
Bacillus subtilis ypgA Gene Is fni, a Nonessential Gene Encoding Type 2 Isopentenyl
Diphosphate Isomerase
Motoki Takagi, Kazuhide Kaneda,* and Tomohisa Kuzuyama†
Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan
We previously identified the fni gene of Streptomyces sp. strain CL190 as type 2 isopentenyl diphosphate (IPP) isomerase, which needs both FMN and NADPH for enzyme activity. An fni gene homolog, ypgA, was detected in the database of the Bacillus subtilis genome. However, the ypgA product was about 140 amino acids shorter in the N-terminal than the Streptomyces fni gene product. A database search found three new putative start codons in 129, 225, and 411 bases upstream of the original start codon of the ypgA gene. The longest gene product, which was named ypgA3, showed the most significant homology to the Streptomyces fni gene product. The ypgA3 gene was expressed with an N-terminal His-tag in Escherichia coli and the purified soluble protein was characterized in detail. The ypgA3 protein converted IPP to its isomer dimethylallyl diphosphate in the presence of both FMN and NADPH. The enzyme also catalyzed the reverse reaction in the presence of both the cofactors. Disruption of the ypgA3 gene was not lethal to B. subtilis. These results indicate that Bacillus ypgA3 gene is fni, a nonessential gene encoding type 2 IPP isomerase.
-19-
Royal Jelly Inhibits the Production of Proinflammatory Cytokines by Activated
Macrophages
Keizo Kohno,1,2,† Iwao Okamoto,1 Osamu Sano,1 Norie Arai,1 Kanso Iwaki,1 Masao Ikeda,1 and Masashi Kurimoto1
1Fujisaki Institute, Hayashibara Biochemical Laboratories, Inc., Fujisaki 675-1, Okayama 702-8006, Japan
2Department of Bioresource Science and Technology, Hiroshima University Graduate School of Biosphere Science, Kagamiyama 1-4-4, Higasihiroshima 739-8528, Japan
In this study, we have examined the anti-inflammatory actions of royal jelly (RJ) at a cytokine level. When supernatants of RJ suspensions were added to a culture of mouse peritoneal macrophages stimulated with lipopolysaccharide and IFN-γ, the production of proinflammatory cytokines, such as TNF-α, IL-6, and IL-1, was efficiently inhibited in a dose-dependent manner without having cytotoxic effects on macrophages. This suggests that RJ contains factor(s) responsible for the suppression of proinflammatory cytokine secretion. We named the factor for honeybees RJ-derived anti-inflammatory factor (HBRJ-AIF), and further investigated the molecular aspects of it. Size fractionation study showed that HBRJ-AIF is composed of substances of low (<5 kDa) and high (>30 kDa) molecular weights, with the former being a major component. Chromatographic analysis showed that MRJP3 is one candidate for the HBRJ-AIF with high molecular weights. Thus, our results suggest that RJ has anti-inflammatory actions through inhibiting proinflammatory cytokine production by activated macrophages.
-20-
Cloning of a Gene Cluster Responsible for the Biosynthesis of Diterpene Aphidicolin,
a Specific Inhibitor of DNA Polymerase α
Tomonobu Toyomasu,1 Kentaro Nakaminami,1 Hiroaki Toshima,2 Takashi Mie,3 Kenji Watanabe,3 Hiroyuki Ito,3 Hirokazu Matsui,3 Wataru Mitsuhashi,1 Takeshi Sassa,1 and Hideaki Oikawa4,†
1Department of Bioresource Engineering, Faculty of Agriculture, Yamagata University, Tsuruoka, Yamagata 997-8555, Japan
2Department of Bioresource Science, College of Agriculture, Ibaraki University, Tsukuba, Ibaraki 305-8642, Japan
3Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
4Division of Chemistry, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan
The fungal diterpene, aphidicolin, is a well-known specific inhibitor of DNA polymerase α. Terpenoids are an important class of natural products. However, identification of the biosynthetic gene cluster in terpenoids is relatively rare compared with another important class of natural products, polyketides. To explore a reliable identification method for the biosynthetic gene cluster in fungal diterpenoids, cloning of the biosynthetic gene cluster of aphidicolin was employed. The application of a simple PCR method for genome walking based on the sequence of cDNA encoding aphidicolan-16β-ol synthase (ACS) allowed us to analyze a 15.6-kb region of the Phoma betae genomic DNA. Six ORFs, PbGGS, ACS, PbP450-1, PbP450-2, PbTP, and PbTF were found in this region, and respectively expected to encode geranylgeranyl diphosphate synthase, diterpene synthase, two cytochrome P-450s, the transporter and transcription factor. Their amino acid sequences and introns were deduced by a corresponding cDNA analysis. This study shows that simple PCR-based genome walking without constructing a genomic DNA library is useful for identification of a small gene cluster. We propose a general strategy for the cloning the biosynthetic genes of fungal diterpenoids by using fungal GGS.
-21-
Chiral Discrimination of Branched-chain Fatty Acids by Reversed-phase HPLC after
Labeling with a Chiral Fluorescent Conversion Reagent
Kazuaki Akasaka and Hiroshi Ohrui†
Graduate School of Life Sciences, Tohoku University, Tsutsumidori-Amamiyamachi 1-1, Aoba-ku, Sendai 981-8555, Japan
Anteiso fatty acids having 16 to 29 carbon atoms were labeled with the chiral fluorescent conversion reagents, (1R,2R)- and (1S,2S)-2-(2,3-anthracenedicarboximido)cyclohexanol. The diastereomeric esters of anteiso acids having up to 20 carbon atoms were separated into two peaks in an ODS column under low column-temperature conditions, while those having more than 21 carbon atoms were not separated. A C30 column made it possible to separate diastereomeric esters up to C29 anteiso acid. It was possible to predict the absolute configuration of each acid by the elution order of the derivatives.
-22-
The Primary Structure of a Novel Goose-type Lysozyme from Rhea Egg White*
Jureerut Pooart, Takao Torikata, and Tomohiro Araki†
Department of Biochemistry, School of Agriculture, Kyushu Tokai University, Aso, Kumamoto 869-1404, Japan
G-type lysozyme is a hydrolytic enzyme sharing a similar tertiary structure with plant chitinase. To discover the relation of function and structure, we analyzed the primary structure of new G-type lysozyme. The complete 185 amino acid residues of lysozyme from rhea egg white were sequenced using the peptides hydrolyzed by trypsin, V8 protease, and cyanogen bromide. Rhea lysozyme had sequence similarity to ostrich, cassowary, goose, and black swan, with 93%, 90%, 83%, and 82%, respectively. The six substituted positions were newly found at positions 3 (Asn), 9 (Ser), 43 (Arg), 114 (Ile), 127 (Met), and 129 (Arg) when compared with ostrich, cassowary, goose, and black swan lysozymes. The amino acid substitutions of rhea lysozyme at subsite B were the same as ostrich and cassowary lysozymes (Ser122 and Met123). This study was also constructed in a phylogenetic tree of G-type lysozyme that can be classified into at least three groups of this enzyme, namely, group 1; rhea, ostrich, and cassowary, group 2; goose, black swan, and chicken, and group 3; Japanese flounder. The amino acid sequences in assembled three α-helices found in this enzyme group (Thammasirirak, S., Torikata, T., Takami, K., Murata, K., and Araki, T., Biosci. Biotechnol. Biochem., 66, 147-156 (2002)) were also highly conserved, so that they were considered to be important for the formation of the hydrophobic core structure of the catalytic site and for maintaining a similar three-dimensional structure in this enzyme group.
-23-
Metofluthrin: A Potent New Synthetic Pyrethroid with High Vapor Activity against
Mosquitoes
Kazuya Ujihara,† Tatsuya Mori, Tomonori Iwasaki, Masayo Sugano, Yoshinori Shono, and Noritada Matsuo
Agricultural Chemicals Research Laboratory, Sumitomo Chemical Co., Ltd., 4-2-1 Takatsukasa, Takarazuka, Hyogo 665-8555, Japan
(1R)-trans-Norchrysanthemic acid fluorobenzyl esters are synthesized and their structure-activity relationships are discussed. These esters show outstanding insecticidal activity against mosquitoes. In particular, the 2,3,5,6-tetrafluoro-4-methoxymethylbenzyl analog (metofluthrin) exhibits the highest potency, being approximately forty times as potent as d-allethrin in a mosquito coil formulation when tested against southern house mosquitoes (Culex quinquefasciatus). Metofluthrin also exhibits a significant vapor action at room temperature.
-24-
Frost-susceptible Protein in Plasma Membranes in Tubers of Helianthus tuberosus
L.
Keita Arakawa,† Mitsuru Hanazaki, and Shizuo Yoshida
Institute of Low Temperature Science, Hokkaido University, N19-W8, Kita-ku, Sapporo 060-0819, Japan
When plasma membranes were prepared from tubers of Helianthus tuberosus L. (Jerusalem artichoke) frozen at a sublethal temperature (-10°C), the levels of some plasma membrane proteins, named frost-susceptible proteins (FSPs), decreased [Uemura, M., et al., Plant Physiol., 80, 187-195 (1986)]. The aim of this study was to characterize the response of FSP120, which is named FSP-3 in a previous report, to freezing treatment by immunoblotting. Levels of FSP120 in the plasma membranes of tubers decreased after sublethal freezing, whereas no degraded products were detected in the microsomes or the soluble fraction. The amount of FSP120 in the crude extract of frozen tubers remained at a comparable level to that of the unfrozen tubers. These results suggest that FSP120 might be released from plasma membranes during freezing treatment of the tubers of Jerusalem artichoke.
-25-
Synthesis and Antioxidant Activity of Oxygenated Furofuran Lignans
Satoshi Yamauchi,1,† Tomoko Ina,1 Takuya Kirikihira,2 and Toshiya Masuda2
1Faculty of Agriculture, Ehime University, Tarumi 3-5-7, Matsuyama, Ehime 790-8566, Japan
2Faculty of Integrated Arts and Sciences, University of Tokushima, Tokushima 770-8502, Japan
Nine furofuran compounds having a different type of oxidation were synthesized from one common intermediate in a short series of steps, and the antioxidant activity was evaluated. It was found that the tertiary hydroxy group on the furofuran ring affected the degree of antioxidant activity and that the structure, except for the phenolic part, was important for the antioxidant activity.
-26-
Preparation of Selenium-enriched Sprouts and Identification of Their Selenium
Species by High-performance Liquid Chromatography-Inductively Coupled Plasma
Mass Spectrometry
Satoru Sugihara, Mariko Kondo, Yuko Chihara, Mami Yuji, Hiroyuki Hattori, and Munehiro Yoshida†
Laboratory of Food and Nutritional Sciences, Department of Biotechnology, Faculty of Engineering, Kansai University, Yamate 3-3-35, Suita, Osaka 564-8680, Japan
Sprouts of several plants (10 families and 28 species) were cultivated in a high selenium environment, and the chemical species of selenium in these selenium-enriched sprouts were identified by using high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS). Cultivation of sprouts of kaiware daikon (type of radish) with 5.0 μg/ml or 10.0 μg/ml of selenium as selenite inhibited the growth. However, no abnormalities in the shape or color were apparent even in the sprouts exposed to 10.0 μg/ml of selenium. The selenium concentration in the sprouts of most plants examined was higher than that from environmental exposure. Among the types of selenium that were accumulated, a large part (69-98%) was extractable in 0.2 M HCl. Chemical analysis of selenium in the HCl extract showed that the main selenium species in all the sprouts examined was Se-methylselenocysteine. In addition to Se-methylselenocysteine, selenomethionine, non-metabolized selenite, γ-glutamyl-Se-methylselenocysteine and an unknown selenium compound were also detected in several high-selenium sprouts. Since higher anticarcinogenic activities of these monomethylated selenoamino acids have been observed, it is anticipated that such selenium-enriched sprouts will be used as a foodstuff for cancer prevention.
-27-
Tomato Paste Fraction Inhibiting the Formation of Advanced Glycation End-products
Tadashi Kiho,1,† Shigeyuki Usui,2 Kazuyuki Hirano,2 Koichi Aizawa,3 and Takahiro Inakuma3
1Gifu Prefectural Institute of Health and Environmental Sciences, 1-1 Naka-fudogaoka, Kakamigahara 504-0838, Japan
2Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502-8585, Japan
3Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nishinasuno-machi, Nasu-gun, Tochigi 329-2762, Japan
A water-soluble and low-molecular-weight fraction (SB) was obtained from tomato paste. The effects of SB on the formation of advanced glycation end-products (AGE) in protein glycation were studied by the methods of specific fluorescence, ELISA and a Western blot analysis, using the anti-AGE antibody after incubating protein with sugar. The results suggest that SB had strong inhibitory activity, in comparison with aminoguanidine as a positive control, and that the inhibitory mechanism of SB differed from that of aminoguanidine to involve trapping of reactive dicarbonyl intermediates in the early stage of glycation. SB contained an antioxidant, rutin, which showed potent inhibitory activity. The results also suggest that rutin chiefly contributed to inhibiting the formation of AGE, and that other compounds in SB may also have been related to the activity.
-28-
Self-cloning Yeast Strains Containing Novel FAS2 Mutations Produce a Higher
Amount of Ethyl Caproate in Japanese Sake
Kazuo Aritomi,1 Isao Hirosawa,2 Hisashi Hoshida,2 Mikio Shiigi,1 Yoshinori Nishizawa,2 Susumu Kashiwagi,1 and Rinji Akada2,†
1Yamaguchi Prefectural Industrial Technology Institute, Asutopia, Ube 755-0151, Japan
2Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Tokiwadai, Ube 755-8611, Japan
Point mutation of Gly1250Ser (1250S) of the yeast fatty acid synthase gene FAS2 confers cerulenin resistance. This mutation also results in a higher production of the apple-like flavor component ethyl caproate in Japanese sake. We mutated the 1250th codon by in vitro site-directed mutagenesis to encode Ala (1250A) or Cys (1250C) and examined cerulenin resistance and ethyl caproate production. The mutated FAS2 genes were inserted into a binary plasmid vector containing a drug-resistance marker and a counter-selectable marker, GALp-GIN11M86. The plasmids were integrated into the wild-type FAS2 locus of a sake yeast strain, and the loss of the plasmid sequences from the integrants was done by growth on galactose plates, which is permissive for loss of GALp-GIN11M86. These counter-selected strains contained either the wild type or the mutated FAS2 allele but not the plasmid sequences, from which FAS2 mutant strains were selected by allele-specific PCR. The FAS2-1250C mutant produced a higher amount of ethyl caproate in sake than FAS2-1250S, while FAS2-1250A produced an ethyl caproate level intermediate between FAS2-1250S and the parental Kyokai no. 7 strain. Interestingly, these mutants only showed detectable cerulenin resistance. These ‘self-cloning’ yeast strains should be acceptable to the public because they can improve sake quality without the presence of extraneous DNA sequences.
-29-
Note
Effects of Several Variable Factors on the Isotope Ratio by HRGC-MS
Masayoshi Sawamura,1,† Atsushi Satake,1,2 Takao Ueno,1 Akitoshi Une,1 and Hiroyuki Ukeda1
1Department of Bioresources Science, Faculty of Agriculture, Kochi University, B-200 Monobe, Nankoku, Kochi 783-8502, Japan
2Nagaoka Perfumery Co., Ltd., 3-30, 1-chome, Itsukaichi City, Osaka 567-0005, Japan
In the isotope ratio (Ir) analysis using GC-MS, several variable factors in sampling incidental to any food analysis were investigated for yuzu fruit. The Irs of ten monoterpene hydrocarbons in yuzu essential oils from each of six fruiting positions of three trees were measured. The sign test following t-test of all the Ir values demonstrated that there was no significant difference between both sampling years of 2001 and 2002. There was also no significant variation in the Ir values among the three trees and six fruiting positions in the individual two years.
-30-
Note
σmarY1, the LTR of the gypsy-Type Retroelement marY1 from the Basidiomycete
Tricholoma matsutake, Allows Multicopy DNA Integration in Lentinula edodes*
Hitoshi Murata† and Yasumasa Miyazaki
Department of Applied Microbiology and Mushroom Sciences, Forestry & Forest Products Research Institute, Tsukuba, Ibaraki 305-8687, Japan
σmarY1 is the LTR of the retroelement marY1 from the homobasidiomycete Tricholoma matsutake. Upon integration through transformation, pLC1-hph carrying a σmarY1 derivative, σmarY1*, conferred the hygromycin-resistant phenotype stronger than the vector without σmarY1* on Lentinula edodes. Based on the densitometric analysis after Southern hybridization, a copy number of the former construct integrated in the genome is much higher than that of the latter. We conclude that σmarY1 allows multicopy DNA integration and will be useful in the genetic research on this fungal group.
-31-
Note
Detection of Hydrolytic Activity of Trypsin with a Fluorescence-chymotryptic
Peptide on a TLC Plate
Tetsuya Uchikoba,1,† Shigeko Fukumoto,1 Takao Itakura,2 Michiko Okubo,3 Kazuhiko Tomokiyo,4 Kazunari Arima,5 and Hiroo Yonezawa5
1Kagoshima University Musium, 1-21-30 Korimoto, Kagoshima 890-0065, Japan
2Department of Biochemistry and Technology of Aquatic Resources, Faculty of Fisheries, Kagoshima University, 50-20 Shimoarata, Kagoshima 890-0065, Japan
3Department of Health and Nutrition, Kagoshima Immaculate Heart University, 2365 Amadatsu-cho, Sendai-shi, Kagoshima 895-0011, Japan
4The Chemo-Sero-Therapeutic Research Institute, 1-6-1 Okubo, Kumamoto 860-0083, Japan
5Laboratory of Biochemistry, Department of Chemistry, Faculty of Science, Kagoshima University, 1-21-35 Korimoto, Kagoshima 890-0065, Japan
To find a new trypsin-like enzyme, a simple assay method of the hydrolysis activity for trypsin has been found. We used 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) in the peptide labeling as a substrate for the trypsin-like peptidase in this study. The peptidase activity of trypsin was detected by using an AQC-chymotryptic peptide (AHP1) obtained from bovine hemoglobin. This showed that the substrate specificity of trypsin-like peptidase was distinguishable from that of the others by this procedure, and the method was used extensively in cases of various trypsin inhibitors with no significant interference from the concomitant.
-32-
Note
SMXA-5 Mouse as a Diabetic Model Susceptible to Feeding a High-fat Diet
Misato Kobayashi,1 Fusayo Io,1 Takahiro Kawai,1 Masahiko Nishimura,2 Tamio Ohno,2 and Fumihiko Horio1,†
1Department of Applied Molecular Bioscience, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan
2Institute for Laboratory Animal Research, Nagoya University School of Medicine, Japan
The SMXA-5 strain, a new mouse model for type 2 diabetes, is a recombinant inbred strain derived from non-diabetic SM/J and A/J strains. As dietary fat is a key component in the development of diabetes, we compared the glucose tolerance and diabetes-related traits among the SMXA-5, SM/J, and A/J strains while feeding a high-fat diet for 10 weeks. SMXA-5 fed on a high-fat diet showed an increased serum insulin concentration. Judging from the hyperinsulinemia in SMXA-5, this strain showed insulin resistance, an inability of peripheral tissues to respond to insulin, which was strengthened by feeding with a high-fat diet. When fed on a high-fat diet for 5 weeks, the SMXA-5 mice showed severely impaired glucose tolerance. On the other hand, SM/J showed mildly impaired glucose tolerance, even when fed on a high-fat diet for 10 weeks. These results indicate that SMXA-5 would be available for use as a diabetic model susceptible to a high-fat diet.
-33-
Note
Distribution of Minerals in Quinoa (Chenopodium quinoa Willd.) Seeds
Yotaro Konishi,1,† Shigeru Hirano,1,2 Hideki Tsuboi,3 and Masao Wada3
1Graduate School of Human Life Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan
2Dai-Nippon Meiji Sugar Co., Ltd., 1-5-3 Nihonbashi, Chuo-ku, Tokyo 103-0027, Japan
3Application Technology Department, Naka Customer Center, Hitachi Science Systems Ltd., 11-1 Ishikawa-cho, Hitachinaka, Ibaraki 312-0057, Japan
The distribution of minerals in quinoa (Chenopodium quinoa Willd.) seed was examined using energy dispersive X-ray microanalysis (EDX) in combination with scanning electron microscopy (SEM). Phosphorus, K, and Mg coincided in localization in embryonic tissue. Since phytin globoids have been known to localize in protein bodies in embryonic cells of quinoa seed, it is thought that P is attributed to phytic acid and that K and Mg form to phytate. Calcium and K were present in the pericarp, where the cell wall is thickly developed, suggesting that these minerals are associated with pectin. Sulfur occurred in embryonic tissues, which would be derived from sulfur amino acid residues of storage proteins concentrated in the tissues. Abrasion of quinoa seeds resulted particularly in decrease in Ca content.
-34-
Note
High-level Production of Hyperthermophilic Cellulase in the Bacillus brevis
Expression and Secretion System
Yasuhiro Kashima1 and Shigezo Udaka2,†
1Special Division for Human Life Technology, National Institute of Advanced Industrial Science and Technology (AIST Kansai), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan
2Nagoya University, Chikusa, Nagoya 464-8601, Japan
A hyperthermophilic cellulase derived from Pyrococcus horikoshii was successfully produced with the Bacillus brevis host-vector system. The production of the recombinant enzyme was increased about 20-fold (to a level of 100 mg per liter) by the insertion of certain amino acid such as alanine and peptides like AEEAADP between the carboxyl end of signal peptide and the N-terminus of the mature cellulase. These recombinant cellulases had the same characteristics as that of the cellulase expressed in Escherichia coli.
-35-
Note
Suppressive Effects of Selected Antioxidants on the Activated Leukocytes-induced
Mutagenesis in the Co-culture Assay Systems
Ha Won Kim,1 Akira Murakami,1 Marshall V. Williams,2 and Hajime Ohigashi1,†
1Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
2Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, 2078 Graves Hall, 333 W. 10th Avenue, Columbus, OH 43210, USA
We recently established a novel co-culture assay system using activated inflammatory cells and AS52 Chinese hamster ovary cells, and demonstrated that reactive oxygen and nitrogen species (RONS) generated from activated inflammatory leukocytes induce mutations in the gpt recorder gene in AS52 cells. In this study, we examined the inhibitory effects of 19 agents with antioxidative properties on RONS generation in cultured inflammatory cells and on mutagenesis in AS52 cells co-cultured with activated inflammatory cells. The results demonstrate that there is a linear correlation between the ability of these agents to suppress RONS production in activated inflammatory cells and to inhibit mutation in AS52 cells.
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Note
High Phosphorus Diet Changes Phosphorus Metabolism Regardless of PTH Action
in Rats
Shin-ichi Katsumata,1 Ritsuko Masuyama,1 Moyuru Koshihara,1 Hiroshi Matsuzaki,2 Mariko Uehara,1 and Kazuharu Suzuki1,†
1Department of Nutritional Science, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan
2Department of Nutrition, Junior college of Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan
In this study, we ascertained whether the parathyroid hormone (PTH) dominantly regulated the effects of high phosphorus (P) intakes on urinary excretion of P and bone metabolism in rats. To maintain serum PTH level equally, parathyroidectomy (PTX) and sham-operated rats were constantly exposed to rPTH(1-34) and fed both control (0.3% P) and high P (1.2% P) diet for 7 days, respectively. Urinary excretions of P and C-terminal telopeptides of type I collagen were significantly increased in both PTX and sham rats by the high P diet. These results suggest that high P diet increased urinary P excretion while promoting bone resorption regardless of PTH-dependent regulation.
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Note
Protective Effect of Blue-M1 against Oxidative Stress on COS-1 Cells
Teruyuki Usui,1 Satomi Shizuuchi,1 Hirohito Watanabe,2 and Fumitaka Hayase1,†
1Department of Agricultural Chemistry, Faculty of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
2Department of Life Sciences, Faculty of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan
Blue-M1 is a blue pigment formed from xylose and glycine in the Maillard reaction. Previous work revealed that Blue-M1 scavenged hydroxyl radicals, and prevented the autoxidation of linoleic acid in vitro. We investigated the protective effect of Blue-M1 for 2,2’-azobis(2-amidino-propane)dihydrochloride (AAPH)-induced toxicity in COS-1 cells.
COS-1 cells were cultured in AAPH containing DMEM medium with or without Blue-M1 at 37°C for 24 h. Blue-M1 decreased the AAPH-induced toxicity in COS-1 cells, and this effect was dose-dependent. Furthermore, COS-1 cells were treated with diphenyl-1-pyrenylphosphine (DPPP), as a reagent for the detection of lipid peroxide, and then were cultured in AAPH containing DMEM medium with or without Blue-M1 at 37°C for 6 h. Blue-M1 prevented the AAPH-induced peroxidation of cell membrane on COS-1 cells, and this effect was also dose-dependent. These results suggest that Blue-M1 prevents the oxidative cell injury. Therefore, Blue-M1 will be an antioxidant, which protect against the oxidative stress in living systems.
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Note
Maturation of Fermented Rice-koji Miso Can Be Monitored by an Increase in Fatty
Acid Ethyl Ester
Shigeo Yamabe,1,† Kentaro Kaneko,2 Hiroko Inoue,3 and Toshichika Takita3
1Tokyo Dietitian Academy, 2-23-11 Ikejiri, Setagaya-ku, Tokyo 154-8544, Japan
2Department of Food Science, Applied life Science, Nippon Veterinary and Animal Science University, 2-27-5 Sakai, Musasino-shi, Tokyo 180-8602, Japan
3Department of Nutritional Science, Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-0054, Japan
A mixture of steamed soybean and boiled rice with seeded Aspergillus oryzae was naturally fermented without addition of yeasts or Lactobacilli, and kept matured for 12 months at room temperature. Chemical analysis of this rice-koji miso sample for lipid changes during maturation showed that triacylglycerol was gradually decomposed into free fatty acid, with distinct formation of fatty acid ethyl ester which, six months after the start of fermentation, came to account for 35.0% of total lipid. The ester was constituted primarily with linoleic acid (ca. 50%) and oleic acid (ca. 20%), no appreciable change in this proportion being observed during maturation. Also, the proportion was unique in that this did not reflect the fatty acid composition in a mixture of the two materials. It is possible to monitor the maturation of the rice-koji miso by following up the increase with time in fatty acid ethyl ester.
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Note
Introduction of DPR, an Enterostatin Fragment Peptide, into Soybean β -Conglycinin
α ’ Subunit by Site-directed Mutagenesis
Yasuyuki Takenaka,1,* Naomi Doyama,1 Nobuyuki Maruyama,2 Shigeru Utsumi,2 and Masaaki Yoshikawa1,†
1Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan
2Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan
DPR, a fragment peptide of enterostatin (VPDPR) having hypocholesterolemic activity, was introduced into the three homologous sites, EPR, DYR, and DPI, in the soybean β -conglycinin α ’ subunit by site-directed mutagenesis. The modified β -conglycinin was expressed in Escherichia coli and recovered in the soluble fraction. After purification on ion-exchange HPLC, the modified β -conglycinin was digested by trypsin to release integrated DPR. The yield of DPR from 1 mole of the modified β -conglycinin was 1.2 mole.
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Note
Penipratynolene, a Novel Nematicide from Penicillium bilaiae Chalabuda
Satoshi Nakahara,1 Miyako Kusano,2 Shozo Fujioka,3 Atsumi Shimada,4 and Yasuo Kimura1,†
1Department of Biological and Environmental Chemistry, Faculty of Agriculture, Tottori University, Tottori-shi 680-8553, Japan
2Umea Plant Science Centre, Department of Forest Genetics and Plant Physiology, The Swedish University of Agricultural Sciences, SE-901 83 Umea, Sweden
3The Institute of Physical and Chemical Research (RIKEN), Wako-shi 351-0198, Japan
4Department of Environmental Chemistry, Faculty of Engineering, Kyushu Kyoritsu University, Kitakyushu-shi 807-8585, Japan
New acetylenic nematicidal compound, penipratynolene (1), methy (2’R)-4-(2’-hydroxy-3’-butynoxy)benzoate, together with two known compounds, 6-methoxycarbonylpicolinic acid (2) and 2,6-pyridinedicarboxylic acid (3), were isolated from the culture filtrate of Penicillium bilaiae Chalabuda. The structures of 1-3 were established by spectroscopic methods. The absolute configuration of 1 was confirmed by using a modified version of Mosher’s method. Compounds 1-3 showed nematicidal activity of 77%, 52%, and 98%, respectively, by a bioassay at 300 mg/l with the root-lesion nematode Pratylenchus penetrans.
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Note
A New Prenylated Flavonoid from Propolis Collected in Okinawa, Japan
Shigenori Kumazawa,1,† Hitomi Goto,1 Tomoko Hamasaka,1 Syuichi Fukumoto,2 Takunori Fujimoto,3 and Tsutomu Nakayama1
1Laboratory of Functional Food Science and COE Program in the 21st Century, School of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan
2Pokka Corporation, Shikatsu-cho, Nishikasugai-gun, Aichi 481-8515, Japan
3Japan Propolis Conference, 6-27-12 Honmachi, Nakano, Tokyo 164-0012, Japan
The new prenylflavonoid, isonymphaeol-B (1), together with three known compounds, nymphaeol-A (2), nymphaeol-B (3), and nymphaeol-C (4), were isolated from propolis collected in Okinawa, the southern-most prefecture of Japan. The structure of each compound was determined by spectral methods, including mass spectrometry and 2D NMR. Each compound had 1,1-diphenyl-2-picryl-hydrazyl radical-scavenging activity.
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Communication
Mac1 Positive Cells Are Required for Enhancement of Splenocytes Proliferation
Caused by Bisphenol A
Masao Goto,† Hiroshi Ono, Yuko Takano-Ishikawa, Hiroshi Shinmoto, and Mitsuru Yoshida
National Food Research Institute, Kannondai, Tsukuba, Ibaraki 305-8642, Japan
We examined the effects of bisphenol A (BPA) on immune cells and it was shown that BPA upregulated the proliferation of murine splenocytes stimulated with Concanavalin A (ConA). The upregulating effects of BPA were removed with depleting Mac1+ cells from the splenocytes. This study provides evidence for the first time that Mac1+ cells were required for enhancement of splenocytes proliferation caused by bisphenol A.
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Communication
Truncated Sla1 Induces Haploid Meiosis through the Pat1-Mei2 System in Fission
Yeast
Kaori Tanabe, Katsunori Tanaka, Hideyuki Matsuda, and Makoto Kawamukai†
Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan
We previously reported that expression of Sla1 Δ C, a truncated form of Sla1, induces ectopic meiosis in heterothallic fission yeast and this was possibly due to the inhibition of Pat1 kinase by Sla1 Δ C. Here we found mei2 mRNA and the Mei2 protein accumulated and stability of the Mei2 protein increased when Sla1 Δ C was expressed. The former two results are considered to be the consequence of de-repression of Ste11, which is the transcription factor of mei2 and negatively regulated by Pat1 kinase. The latter result reflects the consequence of deregulation of Mei2 by Pat1 kinase. In addition, Ste11 accumulated in the nucleus when Sla1 Δ C was expressed. All these data consistently support the idea that the action of Sla1 Δ C is to inactivate Pat1 kinase.