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Production, purification and characterization of D-aspartate oxidase from the fungus Trichoderma harzianumSKW-36

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Advances in Bioscience and Biotechnology 2012年第1期3卷 7-13页

作者： Shigekazu Yano Hiroyuki Ozaki Shinji Matsuo Masahiro Ito mamoru wakayama Department of Biotechnology Faculty of Life Sciences Ritsumeikan University Kusatsu Shiga Japan

Trichoderma harzianum Rifai SKW-36 produced two kinds of D-amino acid oxidizing enzymes. One enzyme was D-aspartate oxidase acting on acidic D-amino acids such as D-aspartate and D-glutamate and another one was D-amino acid oxidase acting on neutral D-amino acid such as D-phenylalanine and D-methionine. These enzymes in the cell-free extract were separated by DEAE-Toyopearl ion-exchange column chromatography. Casamino acids, peptone, and yeast extract as carbon and nitrogen sources were effective for the production of the enzymes. No D-amino acid tested induced the production of the enzymes. Casamino acid (0.33%) as carbon and nitrogen source gave a highest specific activity of D-aspartate oxidase among media tested. D-Aspartate oxidase, which was purified by four-step column chromatography in addition to ammonium sulfate precipitation, exhibited a subunit molecular mass of 40 kDa by SDS-PAGE analysis. D-Aspartate, D-glutamate and N-methyl-D- aspartate were oxidized as substrates with the specific activities of 7.80 U/mg, 4.90 U/mg, and 4.22 U/mg, respectively. D-Asparagine, D-glutamine, D-alanine, and D-valine were slightly oxidized. No other D- amino acids tested were inert. The enzyme exhibited relatively wide substrate specificity compared to D-aspartate oxidases reported so far. The pH and temperature optima were 7.5 - 8.0 and 35°C, respectively. The enzyme was stable at pH 6.0 - 9.0. About 75% of the enzyme activity was retained even after treating the enzyme at 50°C for 10 min. The enzyme activity was inhibited not by benzoate and tartrate, but 60% and 24% by fumarate and malonate, respectively.

关键词： D-Aspartate Oxidase Trichoderma harzianum Purification Characterization

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Efficient Production of L-Theanine Using Immobilized Recombinant Escherichia coliCells Expressing a Modified γ-Glutamyltranspeptidase Gene from Pseudomonas nitroreducens

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Advances in Biological Chemistry 2020年第6期10卷 157-171页

作者： Putthapong Phumsombat Chiharu Sano Hiroki Ikezoe Junji Hayashi Takafumi Itoh Takao Hibi mamoru wakayama Department of Biotechnology College of Life Sciences Ritsumeikan University Shiga Japan Graduate School of Technology Industrial and Social Sciences Tokushima University Tokushima Japan Department of Bioscience and Biotechnology Fukui Prefectural University Fukui Japan

L-Theanine (γ-glutamylethylamide) is a naturally occurring amino acid derivative known to have several beneficial physiological effects as a diet supplement, and to give an umami taste when used as a food additive. The compound is industrially produced by γ-glutamyltranspeptidase from Pseudomonas nitroreducens (PnGGT). Using recombinant PnGGT, we have shown previously that Trp385, Phe417, and Trp525 are key amino acid residues for recognition of acceptor substrates at the PnGGT active site. Here, we demonstrate that a recombinant W525D mutant of PnGGT produces L-theanine from ethylamine and L-glutamine more efficiently than wild-type PnGGT, attributable to an increased ratio of transfer activity to hydrolysis activity. An efficient production of L-theanine was achieved by immobilizing Escherichia coli cells expressing the W525D PnGGT mutant (E. coli-W525D) using 2% alginate as the supporting material. The highest L-theanine production using immobilized E. coli-W525D, representing a conversion rate of 90%, was achieved in optimal reaction conditions of pH 10, 40°C, and a substrate molar ratio of L-glutamine to ethylamine of 1:10. The immobilized E. co

关键词： L-Theanine γ-Glutamyltranspeptidase Pseudomonas nitroreducens Escherichia coli Immobilization

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Implications of Transient Receptor Potential Cation Channels in Migraine Pathophysiology

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Neuroscience Bulletin 2021年第1期37卷 103-116页

作者： mamoru Shibata Chunhua Tang Department of Neurology Keio University School of MedicineTokyo 160-8582Japan Department of Neurology Tokyo Dental College Ichikawa General HospitalChiba 272-8513Japan Department of Neurology and Center for Clinical Neuroscience Daping HospitalThird Military Medical UniversityChongqing 400042China

Migraine is a common and debilitating headache disorder. Although its pathogenesis remains elusive,abnormal trigeminal and central nervous system activity is likely to play an important role. Transient receptor potential(TRP) channels, which transduce noxious stimuli into pain signals, are expressed in trigeminal ganglion neurons and brain regions closely associated with the pathophysiology of migraine. In the trigeminal ganglion,TRP channels co-localize with calcitonin gene-related peptide, a neuropeptide crucially implicated in migraine pathophysiology. Many preclinical and clinical data support the roles of TRP channels in migraine. In particular,activation of TRP cation channel V1 has been shown to regulate calcitonin gene-related peptide release from trigeminal nerves. Intriguingly, several effective antimigraine therapies, including botulinum neurotoxin type A, affect the functions of TRP cation channels. Here, we discuss currently available data regarding the roles of major TRP cation channels in the pathophysiology of migraine and the therapeutic applicability thereof.

关键词： Migraine TRPV1 TRPM8 TRPA1 TRPV4 Calcitonin gene-related peptide Trigeminal ganglion Neurogenic inflammation

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Molecular Basis of Neuroimmune Interaction in an In Vitro Coculture Approach

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Cellular & Molecular Immunology 2008年第4期5卷 249-259页

作者： mamoru Nakanishi Tadahide Furuno School of Pharmacy Aichi Gakuin University 1-100 Kusumoto-choChikusa-kuNagoya 464-8650Japan

A team of researchers from Nagoya, Tokyo and Hamilton developed a unique technique for studying neuroimmune interaction with confocal laser scanning fluorescence microscopy several years ago. It relies on guiding immune and nerve cell interaction by creating an adhesive environment using an in vitro coculture dish. With their technique, they are able to study details of the mechanism of how nerve cells communicate with immune cells （mast cells and T lymphocytes） and vice versa. They showed that nerve-mast cell communication could occur in the absence of an intermediary transducing cell and that the neuropeptide substance P, operating v/a NK-1 receptors, was a soluble factor of this communication. In addition, recently, they showed that ATP which was released from activated mast cells mediated the activation of nerve cells. Further, with their technique, Nagoya＇s group was able to study details of the molecular mechanism of nerve-mast cell interaction. N-cadherin and CADM1 （cell adhesion molecule 1） appeared to mediate attachment and promoted the communication between mast cells and nerves predominantly. It would lead to new therapeutic modalities for diseases based on neuroimmune interaction such as neurogenic inflammation, intestinal bowel diseases, asthma, and autoimmune disorders. Cellular ＆ Molecular Immunology. 2008;5（4）：249-259.

关键词： neuro-immunology coculture confocal microscopy soluble factor adhesion molecule

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Accelerated Gas Carburizing (URX Gas Carburizing)

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材料热处理学报 2004年第5期25卷 378-381页

作者： Yoshikazu Shimosato mamoru Kamitani Hiroyuki Nakatsu Chugai Ro Co. Ltd. Osaka JapanChugai Ro Co. Ltd. Osaka JapanChugai Ro Co. Ltd. Osaka Japan

URX gas Carburizing is an accelerated gas Carburizing method with 50% CO + 50% H2 gas which comes from CFLt gas + CO2 gas. By using this gas composition, the Carburizing reaction rate increases to the maximum and the controllability of carbon potential improves. A carbon mass flow rate is the product of the Carburizing reaction rate multiplied by the difference of carbon percent between carbon potential of the gas and the carbon content of the surface of treated steel parts. We have obtained excellent results from the experimental tests at our laboratory as mentioned bellow. 1) Carburizing time can be shortened by 40% for 0.5 - 0.9 mm effective case depth. 2) Uniform Carburizing case depth 3) Less internal oxidation with the same case depth. We have already developed the new roller hearth type continuous Carburizing furnace and the new URX gas generator .

关键词：气体渗碳内部氧化质量流速反应速率系数

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Validation and Application of Skeletochronology for Age Determination of the Ryukyu Ground Gecko, Goniurosaurus kuroiwae(Squamata: Eublepharidae)

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Asian Herpetological Research 2013年第4期4卷 233-241页

作者： Takaki KURITA mamoru TODA Graduate School of Engineering and Science University of the Ryukyus Tropical Biosphere Research Center University of the Ryukyus

Skeletochronology is a method commonly used for estimating the age of amphibians and reptiles in the wild. However, the number of lines of arrested growth （LAGs） does not necessarily reflect age in some species. We validated the applicability of this method to an endangered eublepharid gecko, Goniurosaurus kuroiwae, then inferred its longevity and age structures in wild populations. We classified young geckos into three groups using previously published data for early growth： Group 1 contained hatchlings before the first winter, Group 2 contained hatchlings after the first win- ter, and Group 3 included yearlings after the second winter. LAG numbers in these groups were then compared. All individuals in Group 1 possessed a single LAG, which was considered as a hatching line. Most individuals in Groups 2 and 3 possessed one and two additional LAGs, respectively （LAG1 and LAG2）, corroborating the notion that LAGs are formed annually. A few geckos exhibited fewer LAGs than expected. Analysis of variations in LAG and marrow cavity diameter demonstrated that in animals with fewer LAGs, endosteal resorption or fusion of hatching line and LAG1 had occurred. LAG2 was never lost by endosteal resorption and was identifiable by its diameter. Thus, the age of adult geckos could be determined by counting LAGs outward from LAG2. Application of this method to wild populations re- vealed that the longevity of this species is not less than 83 months, but that almost all individuals in fragmented habitats die before 50 months, suggesting lower population sustainability in such habitats.

关键词： age structure endosteal resorption hatching line longevity rapprochement Ryukyu Archipelago

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Exchange Rate Instability: Japan's Micro-Macro Experiences and Implications for China

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China & World Economy 2006年第2期14卷 32-44页

作者： mamoru Ishida Advisor Itochu Corporation.

Since 1985, the yen-dollar exchange rates repeatedly fluctuated and climbed to a level that could not be justified by economic fundamentals. The impacts on the Japanese economy were serious and far-reaching. Since 21 July 2005, China has been moving toward a more flexible exchange rate regime. Keeping RMB exchange rates basically stable and providing Chinese industries with means to hedge exchange rate risks are essential for China＇s sound economic development.

关键词： exchange rate stability exchange risks forward exchange market

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Variation in Texture and Lankford Value of 1070 Aluminum Sheet Rolled by Cone-shaped Roll

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Journal of Materials Science & Technology 2013年第2期29卷 175-179页

作者： Yasumasa Chino Xinsheng Huang Kazutaka Suzuki mamoru Mabuchi Materials Research Institute for Sustainable Development National Institute of Advanced Industrial Science and Technology 2266-98 AnagahoraShimo-shidami Moriyama-ku Nagoya 463-8560 Japan Department of Energy Science and Technology Graduate School of Energy Science Kyoto University Yoshida-honmachi Sakyo-ku Kyoto 606-8501 Japan

A rolling with cone-shaped roll, the diameter of which continuously varies along the axial direction, has been proposed as a new shear rolling for controlling the texture of an aluminum alloy sheet. In this study, variations in the texture and Lankford value of a 1070 aluminum sheet rolled by the cone-shaped roll were investigated. Rolling with the cone-shaped roll was found to impose intense shear strain at the edges of the specimen, specifically near the surface. The shear directions in the left and right portions of the specimen were opposite to each other. The surface and middle layer of the specimen rolled by the cone-shaped roll and the reference specimen were characterized by a shear texture and typical recrystallization texture components, respectively. Notably, the specimen rolled by the cone-shaped roll exhibited smaller texture intensity than the reference specimen, especially at the surface, and the shear texture-components were observed at relatively deeper positions. As a result of Lankford value measurements, the specimen rolled with the cone-shape roll exhibited a smaller planar anisotropy than the reference specimen and an average Lankford value close to unity, which are likely due to the texture modifications introduced during rolling with the cone-shaped roll.

关键词： Aluminum Mechanical properties Texture Rolling Shear deformation

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Profile and removal of endocrine disrupting chemicals by using an ER/AR competitive ligand binding assay and chemical analyses

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Journal of Environmental Sciences 2009年第7期21卷 900-906页

作者： Ze-hua Liu mamoru Ito Yoshinori Kanjo Atsushi Yamamoto Department of Urban Engineering Graduate School of Engineering Osaka City University 3-3-138 Sugimoto Sumiyoshi-ku Osaka 558-8585 Japan Osaka City Institute of Public Health and Environmental Sciences 8-34 Tojo-cho Tennoji-ku Osaka 543-0026 Japan

An estrogen receptor （ER）/androgen receptor （AR） ligand competitive binding assay （ER/AR-binding assay） and chemical analyses were used to evaluate the endocrine disrupting chemicals （EDCs） behavior of two municipal wastewater treatment plants （WWTPs）（K and S）. In the influents, estrone （E 1）, androsterone （A）, androstenedione （AD）, BPA （bisphenol A）, NP （nonylphenol） and daidzein （DZ） were detected in high amounts with subsequent 24 h-average concentrations of 350, 1000, 29, 1300, 3900, and 5700 ng/L in K-WWTP and of 310, 620, 59, 1600, 2600, and 8400 ng/L in S-WWTP. The estrogenic （androgenic） activity as 17β-estradiol （E2） equivalents （EEQ） or testosterone （Te） equivalents （TEQ） was consequently 620 ng E2/L （570 ng Te/L） and 580 ng E2/L （800 ng Te/L） for the two WWTPs. The removal efficiencies of the above mentioned sole target chemicals were 51%-100% for K-WWTP and 55.6%-100% for S-WWTP. The removal efficiencies of EEQ were about 73% for both WWTPs, while the removal efficiencies of TEQ were 62.1% for K-WWTP and 98.4% for S-WWTP. In addition, chemical-derived EEQ were about 1.2%-52.4% of those by ER-binding assay for K-WWTP and the corresponding ratios were 1.3%-83.3% for S-WWTP, while chemical derived TEQ were less than 3% of values measured by the AR-binding assay for both WWTPs.

关键词： bioassays ER-binding AR-binding 17β-estradiol equivalents testosterone equivalents

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Current advances in humanized mouse models

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Cellular & Molecular Immunology 2012年第3期9卷 208-214页

作者： Ryoji Ito Takeshi Takahashi Ikumi Katano mamoru Ito Central Institute for Experimental Animals Kawasaki Japan

Humanized mouse models that have received human cells or tissue transplants are extremely useful in basic and applied human disease research. Highly immunodeficient mice, which do not reject xenografts and support cell and tissue differentiation and growth, are indispensable for generating additional appropriate models. Since the early 2000s, a series of immunodeficient mice appropriate for generating humanized mice has been successively developed by introducing the IL-2Rynull gene （e.g., NOD/SCID/7c＂uu and Rag2nu＂ycnun mice）. These strains show not only a high rate of human cell engraftment, but also generate well-differentiated multilineage human hematopoietic cells after human hematopoietic stem cell （HSC） transplantation. These humanized mice facilitate the analysis of human hematology and immunology in vivo. However, human hematopoietic cells developed from HSCs are not always phenotypically and functionally identical to those in humans. More recently, a new series of immunodeficient mice compensates for these disadvantages. These mice were generated by genetically introducing human cytokine genes into NODISCIDI？cn＂u and Rag2nulITcnUll mice. In this review, we describe the current knowledge of human hematopoietic cells developed in these mice. Various human disease mouse models using these humanized mice are summarized.

关键词： animal model immunodeficient mice immunology hematology humanized mice

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