(Vol.68 No.4 2004)
Identification of a Collagen Production-promoting Factor from
an Extract of Royal Jelly and Its Possible Mechanism
Satomi Koya-Miyata,† Iwao Okamoto, Shimpei Ushio, Kanso Iwaki, Masao Ikeda,
and Masashi Kurimoto p.767
Characterization of Sericin Powder Prepared from Citric Acid-degraded
Sericin Polypeptides of the Silkworm, Bombyx Mori
Akira Kurioka,† Fujie Kurioka, and Masayoshi Yamazaki p.774
Effects of a Novel Gaseous Antioxidative System Containing a Rosemary
Extract on the Oxidation Induced by Nitrogen Dioxide and Ultraviolet Radiation
Yoshiro Saito,1,* Akira Shiga,2, Yasukazu Yoshida,1,† Takuya Furuhashi,2
Yoji Fujita,2 and Etsuo Niki1 p.781
Multiplicity of 2,3-Dihydroxybiphenyl Dioxygenase Genes in the
Gram-positive Polychlorinated Biphenyl Degrading Bacterium Rhodococcus rhodochrous
K37
Katsuhiko Taguchi,1 Masaki Motoyama,1 and Toshiaki Kudo1,2,† p.787
Relationship between the Preference for Sake (Japanese rice wine)
and the Movements of Metabolic Parameters Coinciding with Sake Intake
Yasuko Manabe,1,† Hitoshi Utsunomiya,1 Kuniyasu Gotoh,1 Takeyuki Kurosu,1
and Tohru Fushiki2 p.796
Transient Assay System for the Analysis of PR-1a Gene Promoter
in Tobacco BY-2 Cells
Sachiko Ono,1 Tsuneyuki Tanaka,1 Yuriko Watakabe,2 and Kazuyuki Hiratsuka1,†
p.803
Construction and Characterization of Chimeric Proteins Composed
of Type-1 and Type-2 Periplasmic Binding Proteins MglB and ArgT
Kenji Kashiwagi,1,4 Kaoru Fukami-Kobayashi,2 Kiyotaka Shiba,1,4,† and
Ken Nishikawa3 p.808
Alkali- and Halo-tolerant Catalase from Halomonas sp. SK1: Overexpression
in Escherichia coli, Purification, Characterization, and Genetic Modification
Le Huyen Ai THUY,1 Krisana PHUCHAROEN,1 Akira IDENO,2 Tadashi MARUYAMA,3,* and
Takao SHINOZAWA1,† p.814
New Brasiliamide Congeners, Brasiliamides C, D and E, from Penicillium
brasilianum Batista JV-379
Tomoyuki Fujita† and Hideo Hayashi p.820
Effects of Physical Fatigue in Mice on Learning Performance in
a Water Maze
Wataru Mizunoya, Shinichi Oyaizu, Akimori Hirayama, and Tohru Fushiki†
p.827
In Vivo and in Vitro Interactions of the Bombyx mori Chymotrypsin
Inhibitor b1 with Escherichia coli
Ningjia He,1 Yoichi Aso,2 Hiroshi Fujii,1,† Yutaka Banno,1 and Kohji Yamamoto1
p.835
Involvement of Tyrosines at Fucose-binding Sites of Aleuria aurantia
Lectin: Non-equal Response to Site-directed Mutagenesis among Five Sites
Koh Amano,1 Masahiro Fujihashi,2 Akikazu Ando,1 Kunio Miki,2 and Yoshiho Nagata1,†
p.841
Zooxanthellactone, a Novel γ-Lactone-type Oxylipine from
Dinoflagellates of Symbiodinium sp.: Structure, Distribution, and Biological
Activity
Ken-ichi Onodera,1 Takuya Fukatsu,1 Nozomi Kawai,1 Yukio Yoshioka,2 Tetsuji
Okamoto,2 Hideshi Nakamura,1,* and Makoto Ojika1,† p.848
No Detectable Transfer of Dietary Lactoferrin or Its Functional
Fragments to Portal Blood in Healthy Adult Rats
Hiroyuki Wakabayashi,† Hidefumi Kuwata,* Koji Yamauchi, Susumu Teraguchi,
and Yoshitaka Tamura p.853
Preparation and Structural Analysis of Actinidain-processed Atelocollagen
of Yellowfin Tuna (Thunnus albacares)
Koichi Morimoto,† Saori Kunii, Kaori Hamano, and Ben’ichiro Tonomura
p.861
Fractionation and Anti-tumor Activity of the Mycelia of Liquid-cultured
Phellinus linteus
Tomoyuki Nakamura,1,2,† Seiichi Matsugo,2 Yasuyuki Uzuka,2 Sumitaka Matsuo,3
and Hirokazu Kawagishi4,† p.868
Enhanced Resistance to Blast Fungus and Bacterial Blight in Transgenic
Rice Constitutively Expressing OsSBP, a Rice Homologue of Mammalian Selenium-binding
Proteins
Kazutoshi Sawada,1,† Morifumi Hasegawa,2 Lena Tokuda,1 Junko Kameyama,1
Osamu Kodama,2 Takayuki Kohchi,3 Kazuya Yoshida,3 and Atsuhiko Shinmyo3 p.873
Immuno-potentiating Effects of the Antler-shaped Fruiting Body
of Ganoderma lucidum (Rokkaku-Reishi)
Michihiro Kohguchi,1,† Toshio Kunikata,1 Hikaru Watanabe,2 Naoki Kudo,2
Takashi Shibuya,2 Tatsuya Ishihara,1 Kanso Iwaki,1 Masao Ikeda,1 Shigeharu Fukuda,2
and Masashi Kurimoto1 p.881
Inverse Correlation between the Nitrogen Balance and Induction
of Rat Liver Serine Dehydratase (SDH) by Dietary Protein
Ryuhei Kanamoto,1,† Kousuke Fujita,1 Megumi Kumasaki,1 Saeko Imai,1 Makoto
Kotaru,2 Tohru Saeki,1 and Kimikazu Iwami1 p.888
Characterization of Murine Grancalcin Specifically Expressed in
Leukocytes and Its Possible Role in Host Defense against Bacterial Infection
Fengzhi Liu,1 Hiroto Shinomiya,1,† Teruo Kirikae,2 Hajime Hirata,3 and
Yoshihiro Asano1 p.894
Novel Lipoxygenase Inhibitors, Tetrapetalone B, C, and D from
Streptomyces sp.
Toshikazu Komoda, Madoka Kishi, Naoki Abe, Yasumasa Sugiyama, and Akira Hirota†
p.903
RSC Nucleosome-remodeling Complex Plays Prominent Roles in Transcriptional
Regulation throughout Budding Yeast Gametogenesis
Hirofumi Koyama,1 Taka-aki Nagao,1 Tomomi Inai,1 Kohji Miyahara,1 Yasufumi Hayasida,1
Katsuhiko Shirahige,2 and Eiko Tsuchiya1,† p.909
Note
Simple Determination of Trace Amounts of Anionic Surfactants in River Water
by Spectrophotometry Combined with Solid-phase Extraction
Masakazu Tsukamoto,1 Yoshiyuki Murata,1 Mikiro Tada,2 and Yasuaki Shimoishi1,†
p.920
Note
Interaction between a Type-II Dockerin Domain and a Type-II Cohesin Domain from
Clostridium thermocellum Cellulosome
Sadanari Jindou,1 Tsutomu Kajino,2 Minoru Inagaki,1 Shuichi Karita,1 Pierre
Beguin,3 Tetsuya Kimura,1 Kazuo Sakka,1,† and Kunio Ohmiya1 p.924
Note
Effect of the Rate of Temperature Increase on Water Quality during Heating in
Electromagnetic- and Gas-heated Pans
Hiroshi Hiratsuka and Ken Sasaki† p.927
Note
Heterologous Mevalonate Production in Streptomyces lividans TK23
Tomohisa Kuzuyama,1,†,* Tohru Dairi,2 Haruyuki Yamashita,3 Yoshikazu Shoji,3
and Haruo Seto4 p.931
Note
Suppression of Dioxin Mediated Aryl Hydrocarbon Receptor Transformation by Ethanolic
Extracts of Propolis
Yong K. Park,1,† Itsuko Fukuda,2 Hitoshi Ashida,2 Shin Nishiumi,2 Julio
Paredes Guzman,1 Helia H. Sato,1 and Glaucia M. Pastore1 p.935
Note
In Vivo Bioconversion of Tetrahydroisoquinoline by Recombinant Coclaurine N-Methyltransferase
Takashi Morishige, Kum-Boo Choi, and Fumihiko Sato† p.939
Note
A New Bipyrrole and Some Phenolic Constituents in Prunes (Prunus domestica L.)
and Their Oxygen Radical Absorbance Capacity (ORAC)
Shin-ichi Kayano,1,2 Hiroe Kikuzaki,2 Takao Ikami,1 Tomoo Suzuki,1 Takahiko
Mitani,1,* and Nobuji Nakatani2,† p.942
Note
Substrate Specificity of Aminopeptidase from the Mid-gut Gland of the Scallop
(Patinopecten yessoensis)
Hironori Umetsu,1,2,† Mito Arai,1 Toshinori Ota,2 Kaoru Abe,3 Hidemitsu
Uchizawa,3 and Kazuo Sasaki2 p.945
Note
Improved Expression of Novel Red- and Green-emitting Luciferases of Phrixothrix
Railroad Worms in Mammalian Cells
Yoshihiro Nakajima,1,† Takuma Kimura,1 Chie Suzuki,1,2 and Yoshihiro Ohmiya1
p.948
Note
Brassinosteroid Selectively Regulates PIN Gene Expression in Arabidopsis
Ayako Nakamura, Hideki Goda, Yukihisa Shimada,† and Shigeo Yoshida p.952
Note
Zooxanthellamide B, a Novel Large Polyhydroxy Metabolite from a Marine Dinoflagellate
of Symbiodinium sp.
Ken-ichi Onodera,1,2 Hideshi Nakamura,1,2,* Yuichi Oba,1,2 and Makoto Ojika1,†
p.955
Note
Properties of Mycelial Aggregate-specific Lectin of Pleurotus cornucopiae Produced
in Pichia pastoris
Futoshi Sumisa, Naoto Iijima, Akikazu Ando, Maiko Shiga, Kozue Kondo, Koh Amano,
and Yoshiho Nagata† p.959
Note
Thr/Ser-rich Domain of Aspergillus Glucoamylase Is Essential for Secretion
Masatoshi Goto,† Noriko Shinoda, Takuji Oka, Yuka Sameshima, Keisuke Ekino,
and Kensuke Furukawa p.961
Note
3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole Induces Apoptosis and Necrosis with
Activation of Different Caspases in Rat Splenocytes
Takashi Hashimoto,1,2 Takashi Sano,1 Wakana Ito,1 Kazuki Kanazawa,1,2 Gen-ichi
Danno,1,2 and Hitoshi Ashida1,2,† p.964
Communication
Effect of Ethanol on Cell Growth of Budding Yeast: Genes That Are Important
for Cell Growth in the Presence of Ethanol
Shunsuke KUBOTA,1 Ikuko TAKEO,1,2 Kazunori KUME,1 Muneyoshi KANAI,1 Atsunori
SHITAMUKAI,1 Masaki MIZUNUMA,1 Tokichi MIYAKAWA,1 Hitoshi SHIMOI,1,3 Haruyuki
IEFUJI,3 and Dai HIRATA1,† p.968
Communication
Differential Expression of Three Plastidial Sigma Factors, OsSIG1, OsSIG2A,
and OsSIG2B, during Leaf Development in Rice
Koji Kasai,1,* Makiko Kawagishi-Kobayashi,2 Masayoshi Teraishi,3 Yasuhiro Ito,1
Kozo Ochi,1 Kyo Wakasa,2 and Yuzuru Tozawa,3,4 p.973
Communication
Unusual Transcription Regulation of the niaD Gene under Anaerobic Conditions
Supporting Fungal Ammonia Fermentation
Kazuto TAKASAKI,1 Hirofumi SHOUN,2 Akira NAKAMURA,1 Takayuki HOSHINO,1 and Naoki
TAKAYA1,† p.978
-1-
Identification of a Collagen Production-promoting Factor from an Extract of
Royal Jelly and Its Possible Mechanism
Satomi Koya-Miyata,† Iwao Okamoto, Shimpei Ushio, Kanso Iwaki, Masao Ikeda, and Masashi Kurimoto
Fujisaki Institute, Hayashibara Biochemical Laboratories, Inc., 675-1 Fujisaki, Okayama 702-8006, Japan
We have previously shown that royal jelly (RJ) promoted collagen production by skin fibroblasts in the presence of ascorbic acid-2-O-alpha-glucoside (AA-2G). In this study, we purified the honeybee RJ-derived collagen production-promoting factor (HBRJ-CPF) from an alkali-solubilized fraction of RJ by C18 reverse-phase column chromatography. The elution profile by the C18 column chromatography and the molecular mass of the purified HBRJ-CPF material coincided with those of 10-hydroxy-2-decenoic acid (10H2DA). We then examined the collagen production-promoting activities of several commercially available fatty acids contained in RJ. We found that 10H2DA and 10-hydroxydecanoic acid increased the collagen production in a dose-dependent manner. Furthermore, 10H2DA induced the fibroblast cell line, NHDF, to produce transforming growth factor-β1 (TGF-β1) which is an important factor for collagen production. As expected, the collagen production-promoting activity of 10H2DA was neutralized by the anti-TGF-β1 antibody. These result suggest that HBRJ-CPF identified as 10H2DA promoted the collagen production of AA-2G-treated fibroblasts by inducing TGF-β1 production.
-2-
Characterization of Sericin Powder Prepared from Citric Acid-degraded Sericin
Polypeptides of the Silkworm, Bombyx Mori
Akira Kurioka,† Fujie Kurioka, and Masayoshi Yamazaki
Silk Science Research Institute, Hyakunincho 3-25-1, Shinjuku-ku, Tokyo 169-0073, Japan
Acid-degraded sericin powder (AC-SP) was prepared from aqueous solution containing citric acid-degraded sericin polypeptides of Bombyx mori. The morphological and biochemical properties of AC-SP were compared with those of alkali-degraded sericin powder (AL-SP) and hot-water degraded sericin powder (HW-SP). Based on an SEM analysis, AC-SP showed a thin film structure of 10–100 μm with good dispersity while AL-SP and HW-SP had a much larger thin film structure (<500μm). The extract of AC-SP showed stronger trypsin inhibitor activity due to cocoon shell trypsin inhibitor (CSTI-IV) than that of HW-SP. The extract of AL-SP showed no CSTI-IV activity. It was found that AC-SP was a trypsin inhibitor complex powder and that the release of CSTI-IV from AC-SP depended on pH and ion strength. Similar powder materials were obtained when such organic acids as tartaric acid and succinic acid were used. These results suggest that the acid-degraded sericin polypeptides work as a protein matrix to which CSTI-IV may bind ionically.
-3-
Effects of a Novel Gaseous Antioxidative System Containing a Rosemary Extract
on the Oxidation Induced by Nitrogen Dioxide and Ultraviolet Radiation
Yoshiro Saito,1,* Akira Shiga,2, Yasukazu Yoshida,1,† Takuya Furuhashi,2 Yoji Fujita,2 and Etsuo Niki1
1Human Stress Signal Research Center, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan
2Living Environment Systems Laboratory, Mitsubishi Electric Corporation, 5-1-1 Ofuna, Kamakura, Kanagawa 247-8501, Japan
Rosemary is commonly used as a spice and a flavoring agent in food processing. Although the antioxidative properties of its extracts have been investigated, there have been few reports on the volatile components of rosemary. We designed a novel antioxidative system which can generate the volatile constituents in the gaseous phase from a rosemary extract and evaluated the gaseous antioxidative activities against both lipid peroxidation and cell death induced by nitrogen dioxide and ultraviolet radiation. The antioxidative effects of the major volatile components on the oxidation of linoleic acid induced by azo compounds were also investigated in a solution.
The volatile components in the novel antioxidative system suppressed the Jurkat cell death induced by nitrogen dioxide and the intracellular formation of reactive oxygen species in fibroblast cells induced by ultraviolet radiation. 1,8-Cineole among the volatile components exerted an antioxidative effect against the oxidation of linoleic acid in a solution induced by azo compounds and ultraviolet radiation. These data suggest that the volatile constituents of a rosemary extract had antioxidative properties and that gaseous exposure antioxidant is a promising method for promoting health.
-4-
Multiplicity of 2,3-Dihydroxybiphenyl Dioxygenase Genes in the Gram-positive
Polychlorinated Biphenyl Degrading Bacterium Rhodococcus rhodochrous K37
Katsuhiko Taguchi,1 Masaki Motoyama,1 and Toshiaki Kudo1,2,†
1Environmental Molecular Biology Laboratory, RIKEN Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
2Science of Biological Supramolecular Systems, Graduate School of Integrated Science, Yokohama City University, Suehiro, Tsurumi-ku, Yokohama 230-0045, Japan
Rhodococcus rhodochrous K37, a Gram-positive bacterium grown under alkaline conditions, was isolated for its ability to metabolize PCBs. Analysis revealed that it has eight genes encoding extradiol dioxygenase, which has 2,3-dihydroxybiphenyl 1,2-dioxygenase activity, and these genes were designated bphC1 to bphC8. According to the classification of extradiol dioxygenases [Eltis, L. D., and Bolin, J. T., J. Bacteriol., 178, 5930–5937 (1996)], BphC3 and BphC6 belong to the type II enzyme group. The other six BphCs were classified as members of the type I extradiol dioxygenase group. BphC4 and BphC8 were classified into a new subfamily of type I, family 3. Two linear plasmids, 200 kb and 270 kb in size, were found in K37, and the bphC6 and bphC8 genes were located in the 200 kb linear plasmid. Northern hybridization analysis revealed that the bphC1, bphC2, and bphC7 genes were induced in the presence of testosterone, the bphC6 gene was induced by fluorene, and the bphC8 gene was induced by biphenyl. All eight BphC products exhibited much higher substrate activity for 2,3-dihydroxybiphenyl than for catechol, 3-methylcatechol, or 4-methylcatechol.
-5-
Relationship between the Preference for Sake (Japanese rice wine) and the Movements
of Metabolic Parameters Coinciding with Sake Intake
Yasuko Manabe,1,† Hitoshi Utsunomiya,1 Kuniyasu Gotoh,1 Takeyuki Kurosu,1 and Tohru Fushiki2
1National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima 739-0046, Japan
2Laboratory of Nutritional Chemistry, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan
Selection of foods by animals largely depends on their physiological condition. In this paper, we reported how physiological changes in rats after ingestion of sake (Japanese rice wine) affected their preference for different kinds of sake. Rats could discriminate among various kinds of sakes in a two-bottle choice test, even after adjustment of the glucose concentration or alcohol concentration of the sake, suggesting that the choice of a sake by animals was based on more than one ingredient. To identify effect of a rat’s physiological condition on the selection of sake, we monitored the levels of blood glucose, nonesterified fatty acids (NEFA), and ketone bodies in serum in mice after forced intragastric ingestion of sake that had previously been offered to the rats in a two-bottle choice test. Blood glucose levels in mice were not different between the rats fed the palatable and unpalatable sakes, and the NEFA level and ketone level were high in rats fed the unpalatable sake. To further clarify the relationship between physiological condition and preference for sake, rats were offered eight kinds of Junmai-syu, which is made with only rice and no sub-ingredient. Rats could still discriminate among Junmai-syu. Furthermore, with two exceptions, the preference for a sake was significantly correlated with the ketone level. The order in the ratio of blood insulin level to blood glucagon level, which is an indicator of metabolism, was correlated with the order in preference to sake. These results suggest that some physiological factors besides oral stimulation also are important factors in the selection of a sake.
-6-
Transient Assay System for the Analysis of PR-1a Gene Promoter in Tobacco BY-2
Cells
Sachiko Ono,1 Tsuneyuki Tanaka,1 Yuriko Watakabe,2 and Kazuyuki Hiratsuka1,†
1Graduate School of Environment and Information Sciences, Yokohama National University, 79-7 Tokiwadai, Hodogaya-ku, Yokohama 240-8501, Japan
2Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma-shi, Nara 630-0101, Japan
In order to develop a rapid and versatile assay system suitable for the analysis of regulated expression of tobacco pathogenesis-related protein 1a (PR-1a) gene, we investigated the use of the transient gene expression system in tobacco BY-2 cells by microprojectile bombardment. Using dual luciferase assay as a reporter gene expression detection system, we observed significant induction of PR-1a promoter activity by salicylic acid (SA) treatment. On the other hand, treatment with 4-hydroxybenzoic acid (4-HBA) resulted in no detectable increase in luciferase activity. Co-expression of a trans-acting factor, the NPR1/NIM1 protein of Arabidopsis, resulted in the induction of higher expression levels of the PR-1a promoter. These results suggest that the assay system is applicable for the analysis of factors involved in the regulated expression of SA-inducible defense-related genes.
-7-
Construction and Characterization of Chimeric Proteins Composed of Type-1 and
Type-2 Periplasmic Binding Proteins MglB and ArgT
Kenji Kashiwagi,1,4 Kaoru Fukami-Kobayashi,2 Kiyotaka Shiba,1,4,† and Ken Nishikawa3
1Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 1-37-1 Kami-Ikebukuro, Toshima-ku, Tokyo 170-8455, Japan
2Bioresource Information Division, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba-shi, Ibaraki 305-0074, Japan
3Laboratory for Gene-Product Informatics, National Institute of Genetics, 1111 Yada, Mishima, Shizuoka 411-8540, Japan
4CREST, Japanese Science and Technology Agency
The respective type-1 and type-2 periplasmic binding proteins (PBPs) MglB and ArgT are believed to have evolved from a common ancestor into siblings showing topological differences in their main chain connectivity. At first glance, they show similar structure. But, more detailed examination reveals that the chain connectivity of ArgT is more convoluted than that of MglB. Reflecting that complexity, the folding of ArgT is complicated and involves intermediate folds. On the other hand, the folding of MglB is a simple two-state transition. In the present study, we constructed and characterized several chimeras made up of various subdomains of MglB and ArgT with the aim of gaining insight into the evolution of protein folding and protein structure. Although these chimeras did not fold as compactly as their parental proteins, some did exhibit cooperative folding, which suggests that novel proteins with new connectivity and new folding pathways could have emerged at a fairly high rate throughout the evolution of proteins.
-8-
Alkali- and Halo-tolerant Catalase from Halomonas sp. SK1: Overexpression in
Escherichia coli, Purification, Characterization, and Genetic Modification
Le Huyen Ai THUY,1 Krisana PHUCHAROEN,1 Akira IDENO,2 Tadashi MARUYAMA,3,* and Takao SHINOZAWA1,†
1Department of Biological and Chemical Engineering, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan
2Sekisui Chemical Co., Ltd., R&D Division Minase Institute, Mishima-gun, Osaka 618-8589, Japan
3Marine Biotechnology Institute, Kamaishi Laboratories, Kamaishi-shi, Iwate 026-0001, Japan
A catalase gene, ohktA, from an alkali- and halo-tolerant bacterium, Halomonas sp. SK1, on the pKK223-3, was expressed in the catalase-lacking Escherichia coli strain UM2. Highly purified catalase showing a single band on SDS-PAGE was obtained by two liquid chromatography steps on DEAE-Toyopear1 and Chelating-Sepharose Fast Flow. The enzyme, oHktA, shows high catalase activity with a pH optimum at 10, and the activity was stable in 4 M KC1. This enzyme is thermo-sensitive, showing a significant loss of activity within 5 minutes at 37 °C. To modify the stability of the catalase, the addition of domain II of the heat stable Mn catalase from Thermus thermophilus to the C-terminus was made. When coexpressed with a chaperone (PhFKBP29) gene product, peptidyl–prolyl cis–trans isomerase, from a thermophilic bacterium, a chimeric catalase was produced in the soluble fraction. The stability of this catalase in the range of 37°–45 °C was improved and it was stable for more than 1 h at 37 °C.
-9-
New Brasiliamide Congeners, Brasiliamides C, D and E, from Penicillium brasilianum
Batista JV-379
Tomoyuki Fujita† 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
Three new brasiliamide congeners, brasiliamides C, D and E, were isolated from okara fermented with Penicillium brasilianum Batista JV-379. Their structures were elucidated on the basis of spectral data and chemical evidence. NMR spectra of these brasiliamides exhibited a mixture of four or two conformers due to the restricted rotation of an amide bond in a solution. The 1H- and 13C-NMR spectral data were analyzed for a major rotamer at an appropriate temperature, since the signals were broadened at room temperature. Both brasiliamides C and D showed convulsive activity against silkworms with an ED50 value of 400 μg/g of diet, whereas brasiliamide E showed less activity than the others.
-10-
Effects of Physical Fatigue in Mice on Learning Performance in a Water Maze
Wataru Mizunoya, Shinichi Oyaizu, Akimori Hirayama, and Tohru Fushiki†
Laboratory of Nutrition Chemistry, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Oiwakecho, Kitashirakawa, Sakyo, Kyoto 606-8502, Japan
We investigated the effects of physical fatigue produced by swimming exercise on learning the Morris water maze in BALB/c mice. We measured the escape latency in the maze immediately after the swimming exercise. The control group was soaked in the water but not fatigued. For easier tasks, like one with an obvious cue flag, the escape latency was not changed by exercise fatigue. However, escape latency was increased after exercise fatigue for more difficult tasks of spatial learning. These results appear to suggest that physical fatigue impaired learning performance. The effects of swimming exercise fatigue on learning efficiency were then investigated. Mice were continuously fatigued during the spatial learning period. This increased escape latency between the first and third sessions. The results suggest that learning efficiency was impaired by exercise fatigue. This system may be useful for screening new foods used to enhance brain function during exercise.
-11-
In Vivo and in Vitro Interactions of the Bombyx mori Chymotrypsin Inhibitor
b1 with Escherichia coli
Ningjia He,1 Yoichi Aso,2 Hiroshi Fujii,1,† Yutaka Banno,1 and Kohji Yamamoto1
1Laboratories of Insect Genetic Resources, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan
2Protein Chemistry and Engineering, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan
Various chymotrypsin inhibitors occur in the hemolymph of silkworm larvae. Interaction of chymotrypsin inhibitor b1 (CI-b1) with Escherichia coli was examined from the viewpoint of action against invading bacteria. Injection of dead E. coli cells into larva reduced the CI-b1 content of the hemolymph, suggesting in vivo binding of CI-b1 to the outer membrane of the cell. Results from incubation of E. coli in cell-free hemolymph in the presence or absence of lipopolysaccharide indicated that CI-b1 is the only CI bound to E. coli and that it interacts with lipopolysaccharide. CI-b1 formed a complex with lipopolysaccharide in vitro; the value of the dissociation constant was relatively large. Inhibitory activity of CI-b1 changed insignificantly in mixture with lipopolysaccharide. CI-b1 affected the growth of E. coli but never worked lethally. CI-b1 is speculated to be a mediator that scavenges intruding bacteria rather than a direct anti-bacterial factor. This is the first report confirming that CI-b1 is a lipopolysaccharide binding protein.
-12-
Involvement of Tyrosines at Fucose-binding Sites of Aleuria aurantia Lectin:
Non-equal Response to Site-directed Mutagenesis among Five Sites
Koh Amano,1 Masahiro Fujihashi,2 Akikazu Ando,1 Kunio Miki,2 and Yoshiho Nagata1,†
1Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo 271-8510, Japan
2Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
Since the involvement of Tyr residues in the fucose-binding of Aleuria aurantia lectin (AAL) was proved by chemical modification using the Tyr-specific reagent tetranitromethane, site-directed mutagenesis was attempted. Since the tertiary structure of AAL was determined recently to be a six-bladed β-propeller fold, and five fucose-binding sites per subunit were found, based on positions of Tyr residues in the tertiary structure, three classes of mutants were constructed: 1) Tyr on the 2nd β-strand of each blade (β-2 mutants), 2) Tyr or Trp on the 3rd β-strand (β-3 mutants), and 3) Tyr outside of binding sites (other-Y mutants). The mutagenized cDNA was expressed in Escherichia coli as His-tag-AAL, and the hemagglutinating activity was assayed. Among 14 mutants, three β-2 mutants (Y26A, Y79A, and Y181A), and three β-3 mutants (Y92A, W149A, and Y241A) showed decreased activity. These mutated residues resided at Sites 1, 2, and 4, at the same locations relatively in the binding sites. Mutagenesis of Tyr or Trp at the corresponding locations in Sites 3 and 5 did not lead to a reduction in activity. Results indicate that the properties of Sites 1, 2, and 4 are different from those of Sites 3 and 5, and that the contribution of these two sites to the hemagglutination reaction was minor.
-13-
Zooxanthellactone, a Novel γ-Lactone-type Oxylipine from Dinoflagellates
of Symbiodinium sp.: Structure, Distribution, and Biological Activity
Ken-ichi Onodera,1 Takuya Fukatsu,1 Nozomi Kawai,1 Yukio Yoshioka,2 Tetsuji Okamoto,2 Hideshi Nakamura,1,* and Makoto Ojika1,†
1Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
2Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan
A novel fatty acid derivative named zooxanthellactone (ZL) was isolated from several strains of symbiotic microalgae, dinoflagellates of the genus Symbiodinium. The metabolite is structurally related to docosahexaenoic acid (DHA) and seems to be biosynthesized by oxidation and subsequent lactonization. The absolute stereochemistry was determined from the specific rotation of the perhydro derivative. The distribution of ZL within several Symbiodinium isolates was quantitatively analyzed by HPLC techniques and suggested a relationship between the productivity of this metabolite and the Symbiodinium phylogeny. The cytotoxicity of ZL was evaluated by using human squamous cell carcinoma cell lines in comparison with that of DHA and other common fatty acids, suggesting that the long unsaturated chain was important rather than the γ-lactone moiety.
-14-
No Detectable Transfer of Dietary Lactoferrin or Its Functional Fragments to
Portal Blood in Healthy Adult Rats
Hiroyuki Wakabayashi,† Hidefumi Kuwata,* Koji Yamauchi, Susumu Teraguchi, and Yoshitaka Tamura
Nutritional Science Laboratory, Morinaga Milk Industry Co., Ltd., Zama, Kanagawa 228-8583, Japan
We investigated the transfer of dietary bovine lactoferrin (LF) and its functional lactoferricin (LFcin) B-containing fragments to the portal blood of healthy adult rats by using several techniques. After a single administration of 125I-labeled LF, radioactive bands were detected in autoradioluminograms of the portal blood, but similar bands were also observed after the administration of [125I]NaI. Although ovalbumin was detected by ELISA at 3–18 ng/ml in the portal blood plasma after an overnight administration, no LF was detected (≤ 1.5 ng/ml). The antibody-captured ovalbumin fragments, but not the LF fragments, were detected in the plasma by surface-enhanced laser desorption/ionization affinity mass spectrometry (SELDI affinity MS). We finally attempted to detect the LFcin B-containing fragments by SELDI affinity MS with on-chip LFcin B-conversion, but could not detect them (≤ 1 ng/ml) in the portal blood after the LF ingestion. The level of LF or its functional fragments transferred to the portal blood was therefore extremely low, if any.
-15-
Preparation and Structural Analysis of Actinidain-processed Atelocollagen of
Yellowfin Tuna (Thunnus albacares)
Koichi Morimoto,† Saori Kunii, Kaori Hamano, and Ben’ichiro Tonomura
Department of Biotechnological Science, Kinki University, Nishi-mitani 930, Uchita, Naga-gun, Wakayama 649-6493, Japan
Pepsin-hydrolyzed collagen (atelocollagen) is a trimer, consisting of α 1 and α 2 monomers, and shows molecular species corresponding to a monomer, dimer (β chain), and trimer (γ chain) by SDS-polyacrylamide gel electrophoresis. Atelocollagen was purified from yellowfin tuna (Thunnus albacares) by salt precipitation and cation-exchange chromatography. Enzymatic hydrolysis of the atelocollagen by actinidain, a cysteine protease purified from kiwifruit, was analyzed by SDS-polyacrylamide gel electrophoresis. The triple helical structure unique to collagen was retained in the atelocollagen as judged by circular dichroism spectra. The actinidain-processed atelocollagen showed only monomeric α 1 and α 2 chains, with no β and γ chains, by SDS-polyacrylamide gel electrophoresis; nevertheless, it retained the typical triple helical structure. It is suggested that actinidain cleaved the atelocollagen molecule at specific sites on the inside of the inter-strand cross-linking peptides.
-16-
Fractionation and Anti-tumor Activity of the Mycelia of Liquid-cultured Phellinus
linteus
Tomoyuki Nakamura,1,2,† Seiichi Matsugo,2 Yasuyuki Uzuka,2 Sumitaka Matsuo,3 and Hirokazu Kawagishi4,†
1Applied Fungi Institute, IBI Corporation, 7841 Anayama-cyo, Nirasaki-shi, Yamanashi 407-0263, Japan
2Division of Applied Biological Sciences, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu-shi, Yamanashi 400-8511, Japan
3Allied Health Sciences, Department of Pharmacology, Kitasato University, 1-15-1 Kitasato, Sagamihara-shi, Kanagawa 228-8555, Japan
4Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan
All the fractions of Phellinus linteus mycelia showed anti-tumor activity toward solid tumors planted in mice. The highest anti-tumor activity of 81.2% was observed in the protein–glucan complex obtained by precipitating the 24% NaOH extract at pH 6.0. This protein–glucan complex consisted of 39.3% polysaccharide and 49.4% protein. Its 13C- and 1H-NMR data showed that the main glucan part of the complex was simple α-1,3-glucan chains.
-17-
Enhanced Resistance to Blast Fungus and Bacterial Blight in Transgenic Rice
Constitutively Expressing OsSBP, a Rice Homologue of Mammalian Selenium-binding
Proteins
Kazutoshi Sawada,1,† Morifumi Hasegawa,2 Lena Tokuda,1 Junko Kameyama,1 Osamu Kodama,2 Takayuki Kohchi,3 Kazuya Yoshida,3 and Atsuhiko Shinmyo3
1Research Association for Biotechnology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan
2Laboratory of Phytochemical Ecology, College of Agriculture, Ibaraki University, 3-21-1 Chuo, Ami, Ibaraki 300-0393, Japan
3Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101, Japan
The rice Oryza sativa selenium-binding protein homologue (OsSBP) gene encodes a homologue of mammalian selenium-binding proteins, and it has been isolated as one of the genes induced by treating a plant with a cerebroside elicitor from rice blast fungus. The possible role of OsSBP in plant defense was evaluated by using a transgenic approach. Plants overexpressing OsSBP showed enhanced resistance to a virulent strain of rice blast fungus as well as to rice bacterial blight. The expression of defense-related genes and the accumulation of phytoalexin after infection by rice blast fungus were accelerated in the OsSBP overexpressors. A higher level of H2O2 accumulation and reduced activity of such scavenging enzymes as ascorbate peroxidase and catalase were seen when the OsSBP-overexpressing plants were treated with the protein phosphatase 1 inhibitor, calyculin A. These results suggest that the upregulation of OsSBP expression conferred enhanced tolerance to different pathogens, possibly by increasing plant sensitivity to endogenous defense responses. Additionally, the OsSBP protein might have a role in modulating the defense mechanism to biotic stress in rice.
-18-
Immuno-potentiating Effects of the Antler-shaped Fruiting Body of Ganoderma
lucidum (Rokkaku-Reishi)
Michihiro Kohguchi,1,† Toshio Kunikata,1 Hikaru Watanabe,2 Naoki Kudo,2 Takashi Shibuya,2 Tatsuya Ishihara,1 Kanso Iwaki,1 Masao Ikeda,1 Shigeharu Fukuda,2 and Masashi Kurimoto1
1Fujisaki Institute, Hayashibara Biochemical Laboratories, Inc., 675-1 Fujisaki, Okayama 702-8006, Japan
2Amase Institute, Hayashibara Biochemical Laboratories, Inc., 7-7 Amase-minami, Okayama 700-0834, Japan
The immuno-potentiating effects of the antler-shaped fruiting body of Ganoderma lucidum (Rokkaku-Reishi, RR), which has been used as a traditional supplement for human health, were investigated in mice. BALB/c mice were administered orally with RR for 3 days at a dose of 50 mg/kg or 500 mg/kg, and interferon-gamma (IFN-γ) production by splenocytes in response to lipopolysaccharide (LPS) was examined on day 4. The oral administration of 500 mg/kg of RR resulted in a significant increase (p<0.05) in IFN-γ production. Stimulation of splenic adherent cells from these mice with LPS also resulted in a significant increase (p<0.05) in interleukin-12 (IL-12) production compared with that from the control mice, suggesting that splenic macrophages were activated by RR administration. Furthermore, 500 mg/kg of RR administered for 14 days resulted in a significant increase (p<0.05) in IFN-γ production by splenocytes in response to both LPS and concanavalin A (Con A). These results suggest that not only splenic macrophages but also T cells were activated by the long-term treatment with RR in vivo. On the other hand, the production of interleukin-4 (IL-4), which is known as an allergic disease-related cytokine, was not affected by the long-term treatment with RR. Our results suggest that the oral administration of RR resulted in Th1-associated immuno-potentiating activities in vivo.
-19-
Inverse Correlation between the Nitrogen Balance and Induction of Rat Liver
Serine Dehydratase (SDH) by Dietary Protein
Ryuhei Kanamoto,1,† Kousuke Fujita,1 Megumi Kumasaki,1 Saeko Imai,1 Makoto Kotaru,2 Tohru Saeki,1 and Kimikazu Iwami1
1Department of Biological Resource Chemistry, Kyoto Prefectural University, Shimogamo, Sakyo-ku, Kyoto 606-8522, Japan
2Department of Food Science, College of Human Ecology, Koka Women’s University, Nishikyogoku, Ukyo-ku, Kyoto 615-0882, Japan
Rats of different ages (3 to 15-wk-old) were fed on a 25% casein diet for one week, and the nitrogen balance and liver serine dehydratase (SDH, EC 4.2.1.13) activity were then determined. The value for nitrogen balance decreased with the age of the rats, while the liver SDH activity increased. A statistical analysis showed clear inverse correlation between the two factors (R2 = 0.7372, p < 0.01). This result suggests that SDH was induced by response to the amount of surplus amino acids from dietary protein taken beyond the body’s requirement. The increase in SDH activity was accompanied by an increase in the level of SDH mRNA. Since the half-life of this mRNA did not change significantly, the induction was mainly controlled at the level of transcription. In addition, the induction seems not to be related to gluconeogenesis, since the mRNA levels of tyrosine aminotransferase (TAT) and phosphoenolpyruvate carboxykinase (PEPCK), other gluconeogenic enzymes, were not changed under these experimental conditions.
-20-
Characterization of Murine Grancalcin Specifically Expressed in Leukocytes and
Its Possible Role in Host Defense against Bacterial Infection
Fengzhi Liu,1 Hiroto Shinomiya,1,† Teruo Kirikae,2 Hajime Hirata,3 and Yoshihiro Asano1
1Department of Immunology and Host Defenses, Ehime University School of Medicine, Ehime 791-0295, Japan
2International Medical Center of Japan, Tokyo 162-8655, Japan
3Department of Life Science, Himeji Institute of Technology, Hyogo 678-1297, Japan
Such phagocytic leukocytes as macrophages and neutrophils are the key cellular components of innate immunity. The actin cytoskeleton is essential for their recruitment and activation in infected tissues. We have previously identified p65/L-plastin with Ca2+-, calmodulin-, and β-actin-binding domains in macrophages. In order to further investigate the p65/L-plastin-involved cellular functions, we cloned the cDNA for murine grancalcin, a possible binding partner of p65/L-plastin. According to the sequence, grancalcin is a member of the penta-EF-hand protein family. We prepared recombinant (r) grancalcin for functional studies and found that it exhibited Ca2+-dependent precipitation. High-titer antibodies against the protein enabled us to detect intracellular grancalcin. A flow cytometric analysis revealed grancalcin to be highly expressed in macrophages and neutrophils. The protein was particularly abundant in those cells recovered from bacteria-infected sites. Immunohistochemical studies clarified that grancalcin was translocated to the actin cytoskeleton in macrophages upon exposure to bacterial lipopolysaccharide. These findings suggest that grancalcin plays a key role in leukocyte-specific functions that are responsible for host defense.
-21-
Novel Lipoxygenase Inhibitors, Tetrapetalone B, C, and D from Streptomyces sp.
Toshikazu Komoda, Madoka Kishi, Naoki Abe, Yasumasa Sugiyama, and Akira Hirota†
Laboratory of Applied Microbiology, School of Food and Nutritional Sciences, University of Shizuoka, Yada 52-1, Shizuoka 422-8526, Japan
Three novel lipoxygenase inhibitors, tetrapetalone B (2, C28H35NO9), C (3, C26H34NO8), and D (4, C28H36NO10), were isolated from a culture broth of Streptomyces sp. USF-4727 that produced a lipoxygenase inhibitor tetrapetalone A (1) simultaneously. Each chemical structure was revealed by spectroscopic evidence, this suggests that these three compounds are structurally related to 1. They had a tetracyclic skeleton and a β-D-rhodinosyl moiety. Tetrapetalone B, C, and D inhibited soybean lipoxygenase with IC50: 320, 360, and 340 μM respectively.
-22-
RSC Nucleosome-remodeling Complex Plays Prominent Roles in Transcriptional Regulation
throughout Budding Yeast Gametogenesis
Hirofumi Koyama,1 Taka-aki Nagao,1 Tomomi Inai,1 Kohji Miyahara,1 Yasufumi Hayasida,1 Katsuhiko Shirahige,2 and Eiko Tsuchiya1,†
1Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Kagamiyama, Higashi-Hiroshima 739-8530, Japan
2Riken Genomic Science Center, Tsurumi, Yokohama 230-0045, Japan
RSC is a nucleosome-remodeling complex of Saccharomyces cerevisiae essential for growth that can alter histone–DNA interaction by using the energy of ATP hydrolysis. Nps1p/Sth1p is an ATPase subunit of RSC. A mutation in the conserved ATPase domain of Nps1p causes a sporulation defect with decreased expression of early meiotic genes, especially IME2. This defect is partially suppressed by the overexpression of either IME1 or IME2. A homozygous diploid of a novel temperature-sensitive nps1 mutation, nps1-13, harboring amino acid substitutions within the bromodomain, was unable to sporulate. Overexpression of IME, IME2, or both of these genes allowed the completion of meiosis I and meiosis II in nps1-13 but not the formation of mature asci. In nps1-13 carrying YEpIME1, the expression of a group of sporulation-specific genes, which express at the middle stages of sporulation and are required for spore-wall formation, notably diminished, and several late sporulation genes expressed at the early stages of sporulation. These results suggest that Nps1p/RSC plays important roles during the spore development process by controlling gene expression for initiating both meiosis and spore morphogenesis, and ensures proper expression timing of late meiotic genes.
-23-
Note
Simple Determination of Trace Amounts of Anionic Surfactants in River Water
by Spectrophotometry Combined with Solid-phase Extraction
Masakazu Tsukamoto,1 Yoshiyuki Murata,1 Mikiro Tada,2 and Yasuaki Shimoishi1,†
1Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Tsushima-naka, Okayama 700-8530, Japan
2Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka, Okayama 700-8530, Japan
A simple and sensitive specrophotometric method combined with solid-phase extraction (SPE) for the simultaneous determination of sodium linear-dodecylbenzenesulfonate (DBS) and sodium dodecyl sulfate (SDS) is described. The C2 (ethyl group bonded silicagel) cartridge could be repeatedly used more than 500 times for SPE, and it enabled the anionic surfactants to be concentrated by 50-fold. The calibration graph for DBS was linear in the range from 1.6 × 10-8 M to 5.0 × 10-7 M and for SDS from 2.0 × 10-9 M to 3.0 × 10-7 M. The relative standard deviation (n=5) for 5.0 × 10-7 M DBS was 3.1% and for 2.5 × 10-7 M SDS was 1.7%. The proposed method was applied to the simultaneous determination of DBS and SDS in river-water samples.
-24-
Note
Interaction between a Type-II Dockerin Domain and a Type-II Cohesin Domain from
Clostridium thermocellum Cellulosome
Sadanari Jindou,1 Tsutomu Kajino,2 Minoru Inagaki,1 Shuichi Karita,1 Pierre Beguin,3 Tetsuya Kimura,1 Kazuo Sakka,1,† and Kunio Ohmiya1
1Faculty of Bioresources, Mie University, 1515 Kamihamacho, Tsu 514-8507, Japan
2Special Research Laboratory II, Toyota Central R&D Laboratories, Inc., Nagakute, Aichi 480-1192, Japan
3Department of Biotechnology, Pasteur Institute, Rue du Dr. Roux 28, Paris 75724, Cedex 15, France
The interaction between the type-II dockerin domain of the scaffoldin protein CipA and the type-II cohesin domain of the outer layer protein SdbA is the fundamental mechanism for anchoring the cellulosome to the cell surface of Clostridium thermocellum. We constructed and purified a dockerin polypeptide and a cohesin polypeptide, and determined affinity constants of the interaction between them by the surface plasmon resonance method. The dissociation constant (KD) value was 1.8 × 10-9 M, which is a little larger than that for the combination of a type-I dockerin and a type-I cohesin.
-25-
Note
Effect of the Rate of Temperature Increase on Water Quality during Heating in
Electromagnetic- and Gas-heated Pans
Hiroshi Hiratsuka and Ken Sasaki†
Materials Science and Engineering, Graduate School of Engineering, Hiroshima Kokusai Gakuin University, 6-20-1 Nakano, Akiku, Hiroshima 739-0321, Japan
More rapid increases in the pH value and hardness during electromagnetic heating of a pan of water were observed than when the pan was heated by LNG or LPG. The water quality changed universally in several tap water samples across Japan. This quality change was closely correlated with the rate of temperature increase, irrespective of heating by electromagnetic induction, LNG or LPG.
-26-
Note
Heterologous Mevalonate Production in Streptomyces lividans TK23
Tomohisa Kuzuyama,1,†,* Tohru Dairi,2 Haruyuki Yamashita,3 Yoshikazu Shoji,3 and Haruo Seto4
1Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan
2Biotechnology Research Center, Toyama Prefectural University, Toyama 939-0398, Japan
3Basic Research Laboratory, Asahi Denka Co., Ltd., Arakawa-ku, Tokyo 116-8553, Japan
4Department of Applied Biology and Chemistry, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan
Mevalonate is a ubiquitous biosynthetic intermediate of terpenoids and is used as a moisturizer in cosmetics and a chemical for biochemical research. In this study, we have achieved a heterologous production of this useful compound by expression in Streptomyces lividans TK23 of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase genes, which were cloned from Streptomyces sp. strain CL190.
-27-
Note
Suppression of Dioxin Mediated Aryl Hydrocarbon Receptor Transformation by Ethanolic
Extracts of Propolis
Yong K. Park,1,† Itsuko Fukuda,2 Hitoshi Ashida,2 Shin Nishiumi,2 Julio Paredes Guzman,1 Helia H. Sato,1 and Glaucia M. Pastore1
1State University of Campinas, College of Food Engineering (UNICAMP), Department of Food Science, Laboratory of Food Biochemistry, Campinas, SP, Brazil
2Kobe University, Department of Biofunctional Chamistry, Faculty of Agriculture, Rokkodai-cho, Nada-ku, Kobe 657-0013, Japan
Present study demonstrated that the ethanolic extracts of propolis containing higher concentrations of flavonoids suppressed 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced aryl hydrocarbon receptor transformation in a dose-dependent manner. The IC50 values of propolis group 3 and group 12 were 1.2 and 3.6 μg/ml, respectively, indicating that propolis showed stronger antagonistic effects as compared with vegetable extracts.
-28-
Note
In Vivo Bioconversion of Tetrahydroisoquinoline by Recombinant Coclaurine N-Methyltransferase
Takashi Morishige, Kum-Boo Choi, and Fumihiko Sato†
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan
Coclaurine N-methyltransferase from Coptis japonica catalyzes the N-methylation of coclaurine as well as simple tetrahydroisoquinoline. We examined the possibility of converting 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline into its N-methylated product using transgenic Escherichia coli, which expressed recombinant coclaurine N-methyltransferase, without the addition of a methyl-group donor. Transgenic E. coli successfully N-methylated the substrate added to the medium and excreted the product. Limitation of bioconversion by the supply of methyl-group donor is discussed.
-29-
Note
A New Bipyrrole and Some Phenolic Constituents in Prunes (Prunus domestica L.)
and Their Oxygen Radical Absorbance Capacity (ORAC)
Shin-ichi Kayano,1,2 Hiroe Kikuzaki,2 Takao Ikami,1 Tomoo Suzuki,1 Takahiko Mitani,1,* and Nobuji Nakatani2,†
1Research & Development Institute, MIKI Corporation, 12-4, Naruohama 3, Nishinomiya 663-8142, Japan
2Division of Food and Health Sciences, Graduate School of Human Life Science, Osaka City University, 3-138, Sugimoto 3, Sumiyoshi, Osaka 558-8585, Japan
Isolation and structural elucidation of prune constituents were performed and total 10 compounds were determined by NMR and MS analyses. A novel compound was identified to be 2-(5-hydroxymethyl-2’,5’-dioxo-2’,3’,4’,5’-tetrahydro-1’H-1,3’-bipyrrole)carbaldehyde, and 7 phenolic compounds were isolated from prunes for the first time. In addition, antioxidant activity of them was evaluated on the basis of the oxygen radical absorbance capacity (ORAC).
-30-
Note
Substrate Specificity of Aminopeptidase from the Mid-gut Gland of the Scallop
(Patinopecten yessoensis)
Hironori Umetsu,1,2,† Mito Arai,1 Toshinori Ota,2 Kaoru Abe,3 Hidemitsu Uchizawa,3 and Kazuo Sasaki2
1Graduate School of Environmental Science, Aomori University, Kobata, Aomori 030-0943, Japan
2Institute of Bioscience and Biotechnology, Aomori University, Kobata, Aomori 030-0943, Japan
3Aomori Industrial Research Center, Daini-tonyamachi, Aomori 030-0013, Japan
An action for various peptides and a kinetic study for amino acid p-nitroanilides (pNAs) and 4-methylcoumaryl-7-amides (MCAs) were performed with purified aminopeptidase from the mid-gut of the scallop. The enzyme preferred dipeptides having Ala, Met, and Phe in the amino-terminal or the penultimate position from the amino-termini. The catalytic efficiencies, kcat/Km values for Ala-pNA and MCA were the highest in the tested substrates, and those for pNA and MCA substrates having Met or Phe were the next highest. The enzyme was found to be a new alanine-specific aminopeptidase.
-31-
Note
Improved Expression of Novel Red- and Green-emitting Luciferases of Phrixothrix
Railroad Worms in Mammalian Cells
Yoshihiro Nakajima,1,† Takuma Kimura,1 Chie Suzuki,1,2 and Yoshihiro Ohmiya1
1Cell Dynamics Research Group, Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan
2TOYO B-Net Co., 27 Wadai, Tsukuba, Ibaraki 300-4247, Japan
Luciferases are widely used for the quantitative monitoring of gene expression in a variety of organisms. We successfully expressed novel red- and green-emitting luciferases of Phrixothrix railroad worms in mammalian cells in combination with the Kozak sequence and the CAG promoter. The characteristic properties of these luciferases indicate that they are appropriate reporter genes for the simultaneous monitoring of two gene expressions.
-32-
Note
Brassinosteroid Selectively Regulates PIN Gene Expression in Arabidopsis
Ayako Nakamura, Hideki Goda, Yukihisa Shimada,† and Shigeo Yoshida
Plant Science Center, RIKEN, Suehiro 1-7-22, Tsurumi, Yokohama, Kanagawa 230-0045, Japan
When brassinosteroid (BR)-deficient mutant (det2) or wild-type (WT) seedlings were treated with brassinolide (BL), the most active BR, for 3 h, the abundance of PIN4 and PIN7 transcripts decreased, and there were fewer PIN4 and PIN7 transcripts in det2 than in the WT. This suggests that BL selectively regulates the PIN gene in a complex manner.
-33-
Note
Zooxanthellamide B, a Novel Large Polyhydroxy Metabolite from a Marine Dinoflagellate
of Symbiodinium sp.
Ken-ichi Onodera,1,2 Hideshi Nakamura,1,2,* Yuichi Oba,1,2 and Makoto Ojika1,†
1Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan
2CREST, JST (Japan Science and Technology)
Zooxanthellamide B, C128H220N2O53S2, a polyhydroxy secondary metabolite, was isolated from a cultured marine dinoflagellate of the genus Symbiodinium. A detailed 2D NMR analysis revealed the chemical structure as a δ -lactone analogue of zooxanthellamide A, which had previously been isolated from the same dinoflagellate by us. The relative configuration of the δ -lactone moiety was determined by NOE experiments and a coupling constant analysis, and that of other ring systems was found to be the same as zooxanthellamide A by the chemical correlation between zooxanthellamides A and B.
-34-
Note
Properties of Mycelial Aggregate-specific Lectin of Pleurotus cornucopiae Produced
in Pichia pastoris
Futoshi Sumisa, Naoto Iijima, Akikazu Ando, Maiko Shiga, Kozue Kondo, Koh Amano, and Yoshiho Nagata†
Department of Bioresources Chemistry, Faculty of Horticulture, Chiba University, Matsudo, Chiba 271-8510, Japan
cDNA of a mycelial aggregate-specific lectin of Pleurotus cornucopiae was expressed in Pichia pastoris, and the expression product was purified and characterized. The product was functional, and the hemagglutinating activity was inhibited most strongly by the addition of N-acetyl-D-galactosamine as was the native lectin. The native lectin is a glycoprotein having five glycosylation recognition signals, and the expression product showed slightly larger molecular mass than that of the native one due to further glycosylation.
-35-
Note
Thr/Ser-rich Domain of Aspergillus Glucoamylase Is Essential for Secretion
Masatoshi Goto,† Noriko Shinoda, Takuji Oka, Yuka Sameshima, Keisuke Ekino, and Kensuke Furukawa
Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University, Hakozaki 6-10-1, Fukuoka 812-8581, Japan
The recombinant Aspergillus awamori strain carrying the mutant glucoamylase-encoding gene in which the entire Thr/Ser-rich Gp-I domain was deleted abolished secretion of mutant glucoamylase. The transcription of the Bip-encoding bipA was low in the wild type (wt) strain, but elevated in the recombinant strain under the condition of glaA expression. The results indicate that the Gp-I domain is vital for glucoamylase secretion.
-36-
Note
3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole Induces Apoptosis and Necrosis with
Activation of Different Caspases in Rat Splenocytes
Takashi Hashimoto,1,2 Takashi Sano,1 Wakana Ito,1 Kazuki Kanazawa,1,2 Gen-ichi Danno,1,2 and Hitoshi Ashida1,2,†
1Department of Biofunctional Chemistry, Faculty of Agriculture, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan
2Division of Life Science, Graduate School of Science and Technology, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan
A dietary carcinogen, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) at 20 μ M activates caspase-3-like proteases as an apoptotic marker in rat splenocytes. The present study demonstrated 100 μ M Trp-P-1 induced necrosis with activation of caspase-3-like proteases. The activation in necrosis and apoptosis resulted from the activation of caspase-9 and caspase-8, respectively. Thus, Trp-P-1 induces apoptosis and necrosis with the activation of different caspases.
-37-
Communication
Effect of Ethanol on Cell Growth of Budding Yeast: Genes That Are Important
for Cell Growth in the Presence of Ethanol
Shunsuke KUBOTA,1 Ikuko TAKEO,1,2 Kazunori KUME,1 Muneyoshi KANAI,1 Atsunori SHITAMUKAI,1 Masaki MIZUNUMA,1 Tokichi MIYAKAWA,1 Hitoshi SHIMOI,1,3 Haruyuki IEFUJI,3 and Dai HIRATA1,†
1Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8530, Japan
2Chiyonoharu-Shuzo, 3312 Shiwa-bori, Higashi-Hiroshima 739-0269, Japan
3National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima 739-0046, Japan
The budding yeast Saccharomyces cerevisiae has been used in the fermentation of various kinds of alcoholic beverages. But the effect of ethanol on the cell growth of this yeast is poorly understood. This study shows that the addition of ethanol causes a cell-cycle delay associated with a transient dispersion of F-actin cytoskeleton, resulting in an increase in cell size. We found that the tyrosine kinase Swe1, the negative regulator of Cdc28-Clb kinase, is related to the regulation of cell growth in the presence of ethanol. Indeed, the increase in cell size due to ethanol was partially abolished in the SWE1-deleted cells, and the amount of Swe1 protein increased transiently in the presence of ethanol. These results indicated that Swe1 is involved in cell size control in the presence of ethanol, and that a signal produced by ethanol causes a transient up-regulation of Swe1. Further we investigated comprehensively the ethanol-sensitive strains in the complete set of 4847 non-essential gene deletions and identified at least 256 genes that are important for cell growth in the presence of ethanol.
-38-
Communication
Differential Expression of Three Plastidial Sigma Factors, OsSIG1, OsSIG2A,
and OsSIG2B, during Leaf Development in Rice
Koji Kasai,1,* Makiko Kawagishi-Kobayashi,2 Masayoshi Teraishi,3 Yasuhiro Ito,1 Kozo Ochi,1 Kyo Wakasa,2 and Yuzuru Tozawa,3,4
1National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan
2National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba, Ibaraki 305-8518, Japan
3Mitsubishi Kagaku Institute of Life Sciences, 1000 Kamoshida-cho, Aoba-ku, Yokohama 227-8502, Japan
4Cell-Free Science and Technology Research Center, 3 Bunkyo-cho, Matsuyama 790-8577, Japan
We isolated and characterized two rice nuclear genes, OsSIG2A and OsSIG2B, encoding the putative σ-factor of the plastid RNA polymerase. Deduced protein sequences predicted a plastid-localizing signal in the N-terminus and subsequent polypeptides similar to known SIG2 proteins. Gene expression analysis revealed that the OsSIG2A transcript is more abundant than the OsSIG2B transcript in all tissues tested and that both rice SIG2s are expressed from earlier stages of leaf development than that in the case of OsSIG1. These results indicate differential expression of SIG genes in leaf morphogenesis, suggesting the existence of tissue- and stage-specific functions of SIG proteins for transcriptional regulation of chloroplast genes in plant development.
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Communication
Unusual Transcription Regulation of the niaD Gene under Anaerobic Conditions
Supporting Fungal Ammonia Fermentation
Kazuto TAKASAKI,1 Hirofumi SHOUN,2 Akira NAKAMURA,1 Takayuki HOSHINO,1 and Naoki TAKAYA1,†
1Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan
2Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-Ku, Tokyo 113-8657, Japan
The niaD gene of the fungus Aspergillus nidulans encodes an assimilatory nitrate reductase and exogenous ammonium represses its expression. Under anoxic conditions, however, A. nidulans expressed niaD even in the presence of ammonium and used the gene product for dissimilatory nitrate reduction (ammonia fermentation). This transcription regulation mechanism under anaerobiosis is critical for the fungus to ferment ammonium.