Faculty, Department/Institute

• Faculty of Chemistry, Materials and Bioengineering Department of Life Science and Biotechnology

Academic status (qualification)

• Professor Apr. 1,2008

Undergraduate Degrees・University

• University of Tsukuba Second Cluster of CollegeCollege of Agriculture and Forestry 1986 Graduated

Graduate Degrees・University

• Kyoto University Doctor's Degree Program Agricultural Chemistry 1992 Completed

Academic Degrees

• Doctor of Agriculture Jul. 1992 Kyoto University

Homepage Address, E-mail Address

• Homepage Address：http://biomole.life-bio.kansai-u.ac.jp/
• E-mail Address：oikawa@kansai-u.ac.jp

Research fields

Research topics

Research Activities

• In graduated School, I researched the Synthesis and function of selenium-containing peptides, and the result was reported to the international Journal, Proc. Nat1. Acad. Sci. U.S.A. Now I am studying the new aspects of the bimolecular Science and technology, and the results were reported to various international Journals and conferences.

Research Career

• Kansai University/Assistant 1991/4/1～1994年/3/31
• Kansai University/Instructor 1993/4/1～1998年/3/31
• Kansai University/Associate Professer
• Kansai University/Professor

Awards

• May 21,2016

Academic Associations

Intellectual Property Rights

• (Published)

  application number：H10-250931 (Sep. 4,1998)

  application number：2000-078969 (Mar. 21,2000)

Research Publications

Academic presentationOther;;;;OIKAWA,Tadao2021/9～

PapersIn refereedAcademic JournalCo-authored;;OIKAWA,Tadao;2021/8～

PapersIn refereedAcademic JournalCo-authoredYAMANAKA,Kazuya;OZAKI,Ryo;HAMANO,Yoshimitsu;OIKAWA,Tadao2021/6～

Research reportUnrefereedOtherCo-authorOIKAWA,Tadao;;;2021/3/18～

Academic presentationOtherCo-authored;;OIKAWA,Tadao2021/2～

Research reportUnrefereedIn-house publicationSingle-AuthorOIKAWA,Tadao2021～

PatentOIKAWA,Tadao;Wakabayashi, Masaki;Ohta, Hiroko2020/7～

PapersIn refereedAcademic JournalCo-authoredYamanaka, Kazuya;Fukumoto, Hibiki;Takehara, Munenori;Hamano Yoshimitsu;OIKAWA,Tadao2020/6～

PapersIn refereedAcademic JournalCo-authoredYamanaka, Kazuya;Hamano, Yoshimitsu;OIKAWA,Tadao2020/5～

Academic presentationCo-authoredOkajima, Kohei;Yamanaka, Kazuya;Kato, Shiro;OIKAWA,Tadao2020/2～

Research reportUnrefereedIn-house publicationSingle-AuthorOIKAWA,Tadao2020～

PapersUnrefereedAcademic JournalCo-authoredKato, Shiro;OIKAWA,Tadao2019/12～

PapersIn refereedAcademic JournalCo-authoredKato, Shiro;Inagaki, Kenji;OIKAWA,Tadao2019/9～

International academic conferenceCo-authoredOIKAWA,Tadao2019/9～

Academic presentationCo-authored;;;;OIKAWA,Tadao;2019/9～

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Academic presentationCo-authored;;;;OIKAWA,Tadao;2019/3～

PapersUnrefereedAcademic JournalCo-authoredOKAJIMA, Kouhei;OIKAWA,Tadao2018/12～

PapersIn refereedAcademic JournalCo-authored;;OIKAWA,Tadao;;2018/12～

PapersIn refereedAcademic JournalCo-authoredAdachi, Motoyasu;Rumi, Shimizu;KAto, Shiro;OIKAWA,Tadao2018/10～10.1007/s00726-018-2671-y

Academic presentationCo-author2018/6～

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PapersIn refereedAcademic JournalCo-authoredKATO, Shiro;OIKAWA,Tadao2018/3～https://doi.org/10.3389/fmicb.2018.00403

BookMonographCo-authored2018/2/25～

Academic presentationCo-authored2018/2～

PapersIn refereedAcademic JournalCo-authored;OIKAWA,Tadao2018/1～

Research reportUnrefereedOtherSingle-Author2018～

PapersIn refereedAcademic JournalCo-authoredWASHIO, Tsubasa;OIKAWA,Tadao2017/12～

PapersIn refereedAcademic JournalCo-authored2017/12～

PapersUnrefereedAcademic JournalSingle-Author2017/12～

Academic presentationCo-authored2017/12～

PapersIn refereedAcademic JournalCo-authoredKATO, Shiro;OIKAWA,Tadao2017/8～10.1128/ genomeA. 00651- 17

PapersIn refereedAcademic JournalCo-authoredKATO, Shiro;OIKAWA,Tadao2017/8～10.1128/genomeA.00656-17

PapersIn refereedAcademic JournalCo-authoredKATO, Shiro;OIKAWA,Tadao2017/8～10.1128/genomeA.00670-17

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PapersIn refereedAcademic JournalCo-authoredKATO, Shiro;OIKAWA,Tadao2017/2～10.1128/genomeA.00661-17

Research reportUnrefereedOtherSingle-Author2017～

PapersUnrefereedAcademic JournalCo-authored2016/12～

BookIn refereedMonographCo-authored chapterOIKAWA,Tadao2016/10/3～

Academic presentation2016/9/14～

Academic presentation2016/9/14～

PapersIn refereedAcademic JournalCo-authoredWASHIO, Tsubasa;KATO, Shiro;OIKAWA,Tadao2016/7～

Academic presentation2016/6/25～

PapersIn refereedAcademic JournalCo-authored;SATO, Ai;OKAMOTO, Yuko;YAMAUCHI, Takae;KATO, Shiro;YOSHIDA, Masahiro;OIKAWA,Tadao;HATA, Yasuo2016～10.1002/prot.25046

Research reportUnrefereedIn-house publication2016～

International academic conferenceUnrefereedOtherCo-authorKATO, Shiro;Masuda, Yuki;KONISHI, Morichika;OIKAWA,Tadao2015/9/5～

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PapersIn refereedAcademic JournalCo-author2015～

PapersIn refereedAcademic JournalCo-authorOIKAWA,Tadao;Morita, Ayaka2015～

PapersIn refereedAcademic JournalCo-authorKATO, Shiro;MASUDA, Yuki;KONISHI, Moichika;OIKAWA,Tadao2015～

PapersIn refereedAcademic JournalCo-author2015～

Research reportUnrefereedIn-house publicationCo-authorOIKAWA,Tadao2015～

CommentaryUnrefereedAcademic JournalSingle-Author2015～

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International academic conferenceUnrefereedOtherCo-authorOIKAWA,Tadao2014/9/3～

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PapersUnrefereedAcademic JournalCo-authored2014～

PapersIn refereedAcademic JournalCo-authorOIKAWA,Tadao;KATO, Shiro2014～

PapersIn refereedAcademic JournalCo-authorHATA, Yasuo;FUJII, Tomomi;YAMAUCHI, Takae;;OIKAWA,Tadao2014～

PapersIn refereedAcademic JournalCo-author;KOBAYASHI, Kazutaka;;YOSHIDA, Masahiro;OIKAWA,Tadao2014～

PapersIn refereedAcademic JournalCo-authoredYAMAUCHI, Takae;;YOSHIDA, Masahiro;OIKAWA,Tadao;HATA, Yasuo2014～

PapersIn refereedAcademic JournalCo-authoredUCHIDA, Yuki;HAYASHI, Hideyuki;WASHIO, Tsubasa;YAMASAKI, Ryo;KATO, ShIro;OIKAWA,Tadao2014～

Research reportUnrefereedIn-house publicationCo-author2014～

Research reportUnrefereedIn-house publicationCo-author2014～

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Academic presentationInternational coauthorship2013/12/2～

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BookUnrefereedIn-house publicationSingle-AuthorOIKAWA,Tadao2012/6～

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Academic presentationCo-authoredOIKAWA,Tadao2012/3/7～

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PapersUnrefereedAcademic JournalCo-authored2012～

Research reportUnrefereedIn-house publicationCo-authored2012～

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PapersIn refereedAcademic JournalCo-authored2012～

Academic presentationCo-authored2011/11/26～

BookUnrefereedOtherSingle-AuthorOIKAWA,Tadao2011/11/15～

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Academic presentationOtherInternational coauthorship2011/6/11～

BookUnrefereedIn-house publicationSingle-AuthorOIKAWA,Tadao2011/6～

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PapersIn refereedAcademic JournalInternational coauthorship2011～

PapersIn refereedAcademic JournalCo-authored2011～

PapersUnrefereedIn-house publicationCo-authored2011～

Academic presentationOtherSingle-AuthorOIKAWA,Tadao2010/12/18～

Academic presentationOtherCo-authored2010/12/7～2010/12/10

Academic presentationOtherCo-authored2010/12/7～2010/12/10

Academic presentationOtherCo-authored2010/12/7～2010/12/10

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Academic presentationOtherCo-authoredOIKAWA,Tadao2010/10/29～

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Academic presentationOtherCo-authored2010/9/17～2010/9/18

Academic presentationOtherSingle-AuthorOIKAWA,Tadao2010/6/30～

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PapersUnrefereedIn-house publicationCo-authored2010～

PapersUnrefereedIn-house publicationSingle-AuthorOIKAWA,Tadao2010～

PapersIn refereedAcademic JournalCo-authored2010～

PapersIn refereedAcademic JournalCo-authored2010～

PapersIn refereedAcademic JournalCo-authored2010～

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PapersOccurrence of D-serine in rice and characterization of rice serine racemaseIn refereedAcademic JournalCo-authoredY. Gogami;K. Ito;Y. Kamitani;Y. Matsushima;T. OikawaPhytochemistry2009～

Academic presentationOtherSingle-AuthorOIKAWA,Tadao2008/9/20～

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PapersIn refereedOtherCo-authored2008～

Building an International Network Exchange Program of Education and Research for Graduate Course Students in Life and BiotechnologyIn-house publicationCo-authoredT. Oikawa;S. Uesato;H. Tamura;H. Kawahara;Y. Nagaoka;K. Shimoke;T. Tsuchido;L. Eggeling;Volker F. Wendisch;Jochen Buchs;Ratana Rujiravanit;Nazalan Najimudin;Chan Lai KengScience and technology Report of Kansai University50, 83-942008～

PapersAcademic JournalSingle-AuthorOIKAWA,Tadao2008～

PapersBuilding an international newwork exchange program of education and research for graduate course students in life science and biotechnologyIn-house publicationInternational coauthorshipT. Oikawa;S. Uesato;H. Tamura;H. Kawahara;Y. Nagaoka;K. Shimoke;T. Tsuchido;L. Eggeling;V. F, Wendisch;J. Buchs;R. Rujiravanit;N. Najimudin;C. L. KengScience and Technology Reports of Kansai UniversityNo.50, p83-942008～

Academic presentationOtherCo-authoredOIKAWA,Tadao2007/12/11～

PapersA cold-active and thermostable alcohol dehydrogenase of a psychrotorelant from Antarctic seawater, Flavobacterium frigidimaris KUC-1In refereedAcademic JournalCo-authoredT. Kazuoka;T. Oikawa;I. Muraoka;I. Kuroda;K. SodaExtremophiles11, 257-2672007/10/28～An NAD(+)-dependent alcohol dehydrogenase of a psychrotorelant from Antarctic seawater, Flavobacterium frigidimaris KUC-1 was purified to homogeneity with an overall yield of about 20% and characterized enzymologically. The enzyme has an apparent molecular weight of 160k and consists of four identical subunits with a molecular weight of 40k. The pI value of the enzyme and its optimum pH for the oxidation reaction were determined to be 6.7 and 7.0, respectively. The enzyme contains 2 gram-atoms Zn per subunit. The enzyme exclusively requires NAD(+) as a coenzyme and shows the pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of NAD(+). F. frigidimaris KUC-1 alcohol dehydrogenase shows as high thermal stability as the enzymes from thermophilic microorganisms. The enzyme is active at 0 to over 85 degrees C and the most active at 70 degrees C. The half-life time and k (cat) value at 60 degrees C were calculated to be 50 min and 27,400 min(-1), respectively. The enzyme also shows high catalytic efficiency at low temperatures (0-20 degrees C) (k (cat)/K (m) at 10 degrees C; 12,600 mM(-1 )min(-1)) similar to other cold-active enzymes from psychrophiles. The alcohol dehydrogenase gene is composed of 1,035 bp and codes 344 amino acid residues with an estimated molecular weight of 36,823. The sequence identities were found with the amino acid sequences of alcohol dehydrogenases from Moraxella sp. TAE123 (67%), Pseudomonas aeruginosa (65%) and Geobacillus stearothermophilus LLD-R (56%). This is the first example of a cold-active and thermostable alcohol dehydrogenase.Grant-in-Aid for Scientific Research

Academic presentationOtherCo-authored2007/9/26～

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Academic presentationOtherCo-authored2007/9/14～

PapersThe Two Carboxylases of Corynebacterium glutamicum Essential for Fatty Acid and Mycolic Acid SynthesisIn refereedAcademic JournalInternational coauthorshipR. Gande;L.G. Dover;K. Krumbach;G.S. Besra;H. Sahm;T. Oikawa;L. EggelingJ. Bacteriol.189, 5257-52642007/7～The suborder Corynebacterianeae comprises bacteria like Mycobacterium tuberculosis and Corynebacterium glutamicum, and these bacteria contain in addition to the linear fatty acids, unique alpha-branched beta-hydroxy fatty acids, called mycolic acids. Whereas acetyl-coenzyme A (CoA) carboxylase activity is required to provide malonyl-CoA for fatty acid synthesis, a new type of carboxylase is apparently additionally present in these bacteria. It activates the alpha-carbon of a linear fatty acid by carboxylation, thus enabling its decarboxylative condensation with a second fatty acid to afford mycolic acid synthesis. We now show that the acetyl-CoA carboxylase of C. glutamicum consists of the biotinylated alpha-subunit AccBC, the beta-subunit AccD1, and the small peptide AccE of 8.9 kDa, forming an active complex of approximately 812,000 Da. The carboxylase involved in mycolic acid synthesis is made up of the two highly similar beta-subunits AccD2 and AccD3 and of AccBC and AccE, the latter two identical to the subunits of the acetyl-CoA carboxylase complex. Since AccD2 and AccD3 orthologues are present in all Corynebacterianeae, these polypeptides are vital for mycolic acid synthesis forming the unique hydrophobic outer layer of these bacteria, and we speculate that the two beta-subunits present serve to lend specificity to this unique large multienzyme complex.

Academic presentationOtherCo-authored2007/6/7～

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PapersBiochemical and genetic analysis of the γ-resorcylate (2,6-dihydroxybenzoate) catabolic pathway in Rhizobium sp. strain MTP-10005: identification and functional analysis of its gene clusterIn refereedAcademic JournalCo-authoredM. Yoshida;T. Oikawa;H. Obata;K. Abe;H. Mihara;N. EsakiJ. Bacteriol.189, 1573-15812007～We identified a gene cluster that is involved in the gamma-resorcylate (2,6-dihydroxybenzoate) catabolism of the aerobic bacterium Rhizobium sp. strain MTP-10005. The cluster consists of the graRDAFCBEK genes, and graA, graB, graC, and graD were heterologously expressed in Escherichia coli. Enzymological studies showed that graD, graA, graC, and graB encode the reductase (GraD) and oxygenase (GraA) components of a resorcinol hydroxylase (EC 1.14.13.x), a maleylacetate reductase (GraC) (EC 1.3.1.32), and a hydroxyquinol 1,2-dioxygenase (GraB) (EC 1.13.11.37). Bioinformatic analyses suggested that graE, graR, and graK encode a protein with an unknown function (GraE), a MarR-type transcriptional regulator (GraR), and a benzoate transporter (GraK). Quantitative reverse transcription-PCR of graF, which encodes gamma-resorcylate decarboxylase, revealed that the maximum relative mRNA expression level ([5.93 +/- 0.82]x 10(-4)) of graF was detected in the total RNA of the cells after one hour of cultivation when gamma-resorcylate was used as the sole carbon source. Reverse transcription-PCR of graDAFCBE showed that these genes are transcribed as a single mRNA and that the transcription of the gene cluster is induced by gamma-resorcylate. These results suggested that the graDAFCBE genes are responsible as an operon for the growth of Rhizobium sp. strain MTP-10005 on gamma-resorcylate and are probably regulated by GraR at the transcriptional level. This is the first report of the gamma-resorcylate catabolic pathway in an aerobic bacterium.

BookAmino Acid Biosynthesis-pathways, Regulation and Metabolic EngineeringUnrefereedMonographInternational coauthorshipOIKAWA TadaoSpringer, Berlin Heidelberg New York273-2882007～

PapersCrystallization and preliminary x-ray diffraction studies of tetrameric malate dehydrogenase from the novel Antarctic psychrophile Flavobacterium frigidimaris KUC-1Academic JournalCo-authoredT. Fujii;T. OIkawa;I. Muraoka;K. Soda;Y. HataActa Crystallogr. Sect. F Struct. Biol. Cryst. Commun.63, 983-9862007～

PapersAcademic JournalCo-authored2007～

PapersIn refereedAcademic JournalCo-authoredOIKAWA,Tadao;;OIKAWA Tadao2006/12/20～

Academic presentationOtherCo-authored2006/9/9～

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Academic presentationOtherCo-authored2006/9/9～

Alanine racemase of Peudomonas taetrolens NBRC 3460：Cloning and its finction in vivoOtherCo-authoredT. Oikawa;S. Osumi;D. Matsui;Y. Kamitani20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress1P－B－1412006/6/19～20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB CongressKyoto, Japan

A novel metabolic pathway for γ－resorcylate：cloning and expression of its gene cluster from Rhizobium sp. MTP－10005OtherCo-authoredM. Yoshida;H. Obata;T. Oikawa20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress1P－B－0572006/6/19～20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB CongressKyoto, Japan

Functional analysis of Asn265 in alcohol dehydrogenase from psychrotorelant, Flavobacterium frigidimaris KUC －1OtherCo-authoredI. Muraoka;T. Tsuchido;T. Oikawa20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress1P－B－0522006/6/19～20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB CongressKyoto, Japan

Academic presentationOtherCo-authored2006/6/16～

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PapersSynthesis and cancer antiproliferative activity of new histone deacetylase inhibitors: hydrophilic hydroxamates and 2-aminobenzamide-containing derivativesIn refereedAcademic JournalCo-authoredY. Nagaoka;T. Maeda;Y. Kawai;D. Nakashima;T. Oikawa;K. Shimoke;T. Ikeuchi;H. Kuwajima;S. UesatoEuropean Journal of Medicinal Chemistry41(6):697-7082006～Kansai University Special Research Fund

PapersAlanine racemase of alfalfa seedlings (Medicago sativa L.): First evidence for the presence of an amino acid racemase in plantsIn refereedAcademic JournalCo-authoredKazutoshi Ono;Kazuki Yanagida;Tadao Oikawa;Tadashi Ogawa;Kenji SodaPhytochemistry67(12):856-8602006～

PapersCrystal Structure of Nonoxidative Zinc-dependent 2,6-Dihydroxybenzoate (gamma-Resorcylate) Decarboxylase from Rhizobium sp. Strain MTP-10005In refereedAcademic JournalCo-authoredMasaru Goto;Hideyuki Hayashi;Ikuko Miyahara;Ken Hirotsu;Masahiro Yoshida;Tadao OikawaJournal of Biokogical Chemistry281, 34365-343732006～

PapersExpression of alr gene from Corynebacterium glutamicum ATCC 13032 in Escherichia coli and molecular characterization of the recombinant alanine racemaseIn refereedAcademic JournalInternational coauthorshipTadao Oikawa;Andreas Tauch;Steffen Schaffer;Toru FujiokaJournal of Biotechnology125, 503-5122006～

PapersMolecular and biochemical characterization of a serine racemase from Arabidopsis thalianaIn refereedAcademic JournalCo-authoredY. Fujitani;N. Nakajima;K. Ishihara;;K. Ito;M. SugimotoPhytochemistry67: 668-6742006～

PatentsCo-authored2006～

PapersCrystal structures of nonoxidative Zn－dependent 2, 6－dihydroxybenzoate（gamma－resorcylate） decarboxylase from Rhizobium sp. strain MTP－10005Academic JournalCo-authoredM. Goto;H. Hayashi;I. Miyahara;K. Hirotsu;M. Yoshida;T. OikawaJournal of Biological Chemistry281, 34365－343732006～

PapersAcademic JournalCo-authored2006～

ConferenceThermostable and cold-active alcohol dehydrogenase from Flavobacterium frigidimaris KUC-1：x-ray crystal structural analysisOtherCo-authoredI. Muraoka;T. Tsuchido;T. OikawaInternational Interdisciplinary Conference on Vitamins， Coenzymes， and Biofactors　2005P0462005/11/10～International Interdisciplinary Conference on Vitamins， Coenzymes， and Biofactors　2005Awaji， Japan

Academic presentationAmino acids racemase with low substrate specificity of Peudomonas putida IFO 12996: Its localization and exportOtherCo-authoredKamitani;D.Matsui;T. OikawaVol.77， No.8， p.813， (3P-220)，2005/10/21～Kobe

Academic presentationThe enzymological studies on recombinant L-lysine：2-oxoglutarate 6-aminotransferaseOtherCo-authoredY. Kashiwabara;Y. Uchida;K. Soda;T.Fujii;T. Narita;H. Agematsu;N. Agata;K. Isshiki;T. OikawaVol.77， No.8， p.813， (3P-221)2005/10/21～Kobe

Academic presentationStructural analysisi of thermostable and cold-active alcohol dehydrogenase from Flavobacterium frigidimaris KUC-1OtherCo-authoredI. Muraoka;A. Kashima;S. Sugio;T. OikawaVol,77, No.8, p.813, (2P435(WS9-8))2005/10/20～Kobe

Academic presentationGenetical analysisi of γ-resorcylate degradation pathway in Rhizobium sp. MTP-10005OtherCo-authoredM. Yoshida;T. OikawaVol.77， No.8，p.818, (2P468(WS9-10))2005/10/20～Kobe

ConferenceThermophilic, Reversible γ-Resorcylate Decarboxylase from Rhizobium sp. MTP-10005: Purification and Molecular CharacterizationIn refereedOtherCo-authoredT. Oikawa;M. YoshidaTagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMMSP0172005/9/26～Tagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMGoettingen, Germany

ConferenceArginine and alanine racemases of Pseudomonas taetrolens IFO 3460: construction of the integrated mutants and their functional characterizationOtherCo-authoredD. Matsui;A. Tauch;K. Krumbach;L. Eggeling;H. Sahm;T. OikawaTagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMMSP0162005/9/26～Tagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMGoettingen, Germany

ConferenceMycolic acid synthesis in Corynebacterium glutamicumOtherCo-authoredR. Gande;G.S. Besra;T. Oikawa;L. EggelingTagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMMSV0092005/9/26～Tagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAMGoettingen, Germany

Academic presentationOtherCo-authored2005/9/20～

Academic presentationOtherCo-authored2005/5/27～

Academic presentationOtherCo-authored2005/5/20～

PapersCo-authored2005/3～

PapersFlarobacterium frigidimaris sp.nov., Isolated from Antarct'c SeawaterIn refereedCo-authoredY. Nogi;K. Soda;T. OikawaSystematic and Applied Microbiology2005～

PapersPurification, Characterization, and Overexpression of Psychrophilic and Thermolabile Malate Dehydrogenase of a Novel Antarctic Psychrotolerant, Flavobacterium frigidimaris KUC-1In refereedAcademic JournalCo-authoredTadao OIkawa;Noriko Yamamoto;Koji Shimoke;Shinichi Uesato;Toshihiko Ikeuchi;Toru FujiokaBiosci. Biotechnol. Biochem.692005～

PapersNGF-induced phosphatidylinositol 3-kinase signaling pathway prevents thapsigargin-triggered ER stress-mediated apotpsis in PC12 cellsIn refereedAcademic JournalCo-authoredKoji Shimoke;Soichiro Kishi;Takahiro Utumi;Yuichi Shimamura;Harue Sasaya;Tadao Oikawa;Shinichi Uesato;Toshihiko IkeuchiNeuroscience Letters389, 124-1282005～Kansai University Special Research Fund

Academic presentationOtherCo-authored2005～

Academic presentationOtherCo-authored2005～

PapersFlavobacterium frigidimaris sp. nov., isolated from Antarctic seawaterIn refereedAcademic JournalCo-authoredY. Nogi;K. Soda;T. OikawaSyst. Appl. Microbiol.28: 310-3152005～

PapersNGFinduced phosphatidylinositol 3-kinase signaling pathway prevents thapsigargin-triggered ER stressmediated apoptosis in PC12 cellsIn refereedAcademic JournalCo-authoredK. Shimoke;S. Kishi;T. Utsumi;Y. Shimamura;H. Sasaya;T. Oikawa;S. Uesato;T. IkeuchiNeuroscience Letters389, 124-1282005～

PapersPurification， characterization， and overexpression of psychrotrophic and thermolabile malate dehydrogenase of a novel Antarctic psychrotolerant, Flavobacterium frigidimaris KUC-1.In refereedAcademic JournalCo-authoredT. Oikawa;N. Yamamoto;K. Shimoke;S. Shimoke;S. Uesato;T. Ikeuchi;T. FujiokaBiosci. Biotech. Biochem69: 2146-21542005～

PapersIn refereedAcademic JournalCo-authoredTrace Nutrients Research22: 39-442005～

PapersIn refereedCo-authoredOIKAWA,Tadao;;;OIKAWA Tadao2004/12～

ConferenceCharacteristics of Cold‐active Enzymes from a Psychrophile from Antarctic seawaterCo-authoredK. Soda;T. Kazuoka;I. Muraoka;Y. Yamanaka;T. Oikawa;S. Sugio2004/10/2～Japan‐Finland Joint Seminar on New Aspects in Microbial BiotechnologyShiran‐Kaikan,Kyoto, Japan.

PapersThermophilic, Reversible γ-Resorcylate Decarboxylase from Rhizobium sp. MTP-10005: Purification, Molecular Characterization, and ExpressionIn refereedCo-authoredM. Yoshida;N. Fukuhara;T. OikawaJ. Bacteriol186 (20), 6855-68632004/10～

PapersAcyl-CoA Carboxylases (accD2 and accD3) together with a Unique Polyketide Synthase (Cg-PKS) are Key to Mycolic Acid Biosynthesis in Corynebacterianeae like Corynebacterium glutamicum and Mycobacterium tuberculosisIn refereedInternational coauthorshipR. Gande;K. J. C. Gibson;A. K. Brown;K. krumbach;L. G. Dover;H. Sahm;S. Shioyama;T. Oikawa;G. Besra;L. EggelingJ. Biol. Chem2004/8～

ConferenceThrmostable NAD-dependent Alcohol Dehydrogenase from a PsychrophileCo-authoredK. Soda;T. Kazuoka;I. Muraoka;T. Oikawa;S. Sugio2004/5/15～2004/5/18The 3rd China-Japan International Conference on VitaminsGuilin, China

Research reportUnrefereedOtherCo-authoredOIKAWA,Tadao;OIKAWA Tadao2004～

Academic presentationOtherCo-authored2004～

Academic presentationAcademic JournalCo-authored2004～

PapersAcyl‐CoA carboxylases (accD2 and accD3) together with a unique polyketide synthase (Cg‐pks) are key to mycolic acid biosynthesis in Corynebacterianeae like Corynebacterium glutamicum and Mycobacterium tuberculosisIn refereedAcademic JournalInternational coauthorshipR. Gande;K. J. C. Gibson;A. K. Brown;K. Krumbach;L. G.Dover;H. Sahm;Susumu Shioyama;Tadao Oikawa;G. S. Besra;L. EggelingJ. Biol. Chem279, 44847‐448572004～

Academic presentationThermophilic, Reversible γ ‐Resorcylate Decarboxylase from Rhizobium sp. MTP‐10005. Purification, Molecular Characterization, and Primary StructureAcademic JournalCo-authoredM. Yoshida;N. Fukuhara;T. OikawaBiochemistryVol.76, No.8, 8252004～Yokohama

Academic presentationZn2+‐Guanidinobutyrase of Arthrobacter sp.KUJ8602:construction of high expression system in Escherichia coliAcademic JournalCo-authoredY. Gougami;I. Muraoka;K. Soda;T. OikawaBiochemistryVol.76, No.8, 9102004～Yokohama

PapersParadoxical thermostable enzymes from psychrophile: molecular characterization and potentiality for biotechnological applicationIn refereedAcademic JournalCo-authoredT. Oikawa;T. Kazuoka;K. Sodabiotechonological application;Thermostable enzymeJ. Molecular Catalysis B: Enzymatic2003/9～NAD(P)+-dependent aldehyde dehydrogenase (EC 1.2.1.5) and aspartase(EC4.3.1.1) in the cells of an atypical psychrophile from Antarctic seawater, Cytophaga sp. KUC-1, were paradoxically thermostable, although they derived from a psychrophile. Both enzymes showed the highest activity at about 55℃, and also active even under cold conditions. The enzymes contained more Ile residues than the enzymes from mesophiles. The Ile/Ile + Val +Leu ratio of the Cytophaga thermostable enzymes was much higher than that of the enzymes from mesophiles. As compared with the enzymes from other microorganisms, the Cytophaga thermostable enzymes have the structural differences in the C-terminal region of the enzymes. Therefore, the C-terminal region might be important for the paradoxical thermostability of the enzymes. The psychorophilic microorganism produces not only psychophilic enzyme, but thermostable enzyme with psychrophilicity. Therefore, the psychrohilic microorganism is one of the candidates for isolation of novel biocatalysts, which have potential for various industrial applications.

PapersNovel Psychrophilic and Thermolabile L-Threonine Dehydrogenase from Psychrophilic Cytophaga sp. Strain KUC-1.In refereedAcademic JournalCo-authoredT. Kazuoka;S. Takigawa;N. Arakawa;Y. Hizukuri;I. Muraoka;T. Oikawa;K. SodaCytophaga;L-TheroninedehydrogenaseJ. Bacteriology2003/8～A psychrophilic bacterium, Cytophaga sp. strain KUC-1, that abundantly produces a NAD+-dependent L-threonine dehydrogenase was isolated from Antarctic seawater, and the enzyme was purified. The molecular weight of the enzyme was estimated to be 139,000, and that of the subunit was determined to be 35,000. The enzyme is a homotetramer. Atomic absorption analysis showed that the enzyme contains no metals. In these respects, the Cytophaga enzyme is distinct from other L-threonine dehydrogenases that have thus far been studied. L-Threonine and DL-threo-3-hydroxynorvaline were the substrates, and NAD+ and some of its analogs served as coenzymes. The enzyme showed maximum activity at pH 9.5 and at 45℃.The kinetic parameters of the enzyme are highly influenced by temperatures. The K m for L-threonine was lowest at 20℃. Dead-end inhibition studies with pyruvate and adenosine-5'-diphosphoribose showed that the enzyme reaction proceeds via the ordered Bi Bi mechanism in which NAD+ binds to an enzyme prior to L-threonine and 2-amino-3-oxobutyrate is released from the enzyme prior to NADH. The enzyme gene was cloned into Escherichia coli , and its nucleotides were sequenced. The enzyme gene contains an open reading frame of 939 bp encoding a protein of 312 amino acid residues. The amino acid sequence of the enzyme showed a significant similarity to that of UDP-glucose 4-epimerase from Staphylococcus aureus and belonging to the short-chain dehydrogenase-reductase superfamily. In contrast, L-threonine dehydrogenase from E. coli belongs to the medium-chain alcohol dehydrogenase family, and its amino acid sequence is not at all similar to that of the Cytophaga enzyme. L-Threonine dehydrogenase is significantly similar to an epimerase, which was shown for the first time. The amino acid residues playing an important role in the catalysis of the E. coli and human UDP-glucose 4-epimerases are highly conserved in the Cytophaga enzyme, except for the residues participating in the substrate binding.

PapersThermostable Aspartase from a Marine Psychorophile, Cytophaga sp.KUC-1: Molecular Characterization and Primary StructureIn refereedAcademic JournalCo-authoredT. Kazuoka;Y. Masuda;T. Oikawa;K. Sodapsychrophile;Aspartase;Cytophaga;Thermostable enzymeJ.Biochem2003/1～We found that a psychrophilic bacterium isolated from Antarctic seawater, Cytophaga sp. KUC-1, abundantly produces aspartase [EC4.3.1.1], and the enzyme was purified to homogeneity. The molecular weight of the enzyme was estimated to be 192,000, and that of the subunit was determined to be 51,000: the enzyme is a homotetramer. L-Aspartate was the exclusive substrate. The optimum pH in the absence and presence of magnesium ions was determined to be pH 7.5 and 8.5, respectively. The enzyme was activated cooperatively by the presence of L-aspartate and by magnesium ions at neutral and alkaline pHs. In the deamination reaction, the K mvalue for L-asparatate was 1.09 mM at pH 7.0, and the S 1/2 value was 2.13 mM at pH 8.5. The V max value were 99.2 U/mg at pH 7.0 and the 326 U/mg at pH8.5. In the amination reaction, the Km values for fumarate and ammonium were 0.797 and 25.2 mM, respectively, and Vmax was 604 U/mg. The optimum temperature of the enzyme was 55℃. The enzyme showed higher pH and thermal stabilities than that from mesophile: the enzyme was stable in the pH range of 4.5-10.5, and about 80% of its activity remained after incubation at 50℃ for 60min. The gene encoding the enzyme was cloned into Escherichia coli , and its nucleotides were sequenced. The gene consisted of an open reading frame of 1,410-bp encoding a protein of 469 amino acid residues. The amino acid sequence of the enzyme showed a high degree of identity to those of other aspartases, although these enzymes show different thermostabilities.

PapersD-Arginase of Arthrobacter sp, KUJ8602: Characterization and Its Identity with Zn2+-Guanidinobutyrase.In refereedAcademic JournalCo-authoredN. Arakawa;M. Igarashi;T. Kazuoka;T. Oikawa;K. SodaZn2+-Guanidinobutyrase;Arthrobacter;D-ArginaseJ. Biochem2003/1～D-Arginase activity was found in the cells of isolate, Arthrobacter sp. KUJ 8602, grown in the L-arginine medium, and the enzyme was purified and characterized. Its molecular weight was estimated to be about 232,000 by gel filtration, and that of the subunit was approximately 40,000 by SDS-PAGE, suggesting that the enzyme is a homohexamer. The enzyme acted on not only D-arginine but also 4-guanidinobutyrate, 3-guanidinopropionate and even L-arginine. The V max/K m values for 4-guanidinobutyrate and D-arginine were determined to be 87 and 0.81 mol/min/mg/mM, respectively. Accordingly, the enzyme is regarded as a kind of guanidinobutyrase [EC3.5.3.7]. The pH optima for 4-guanidinobutyrate and D-arginine were 9.0 and 9.5, respectively. The enzyme was inhibited competitively by 5-aminovalerate, and thiol carboxylates such as mercaptoacetate served as strong mixed-type inhibitors. The enzyme contained about 1 g-atom of firmly bound Zn2+ per mol of subunit, and removal of the metal ions by incubation with 1,10-phenanthroline resulted in loss of activity. The inactivated enzyme was reactivated markedly by incubation with either Zn2+ or Co2+, and slightly by incubation with Mn2+. The nucleotide sequence of enzyme contains an open reading frame that encodes a polypeptide of 353 amino acid residues (M r: 37,993). The predicted amino acid sequence contains sequences involved in the binding of metal ions and the guanidino group of the substrate, which show a high homology with corresponding sequences of Mn2+-dependent amidinohydrolases such as agmatinase from Escherichia coli and L-arginase from rat liver, though the homology of their entire sequences is relatively low (24-43%).

PapersIn refereedAcademic JournalCo-authoredOIKAWA,Tadao;;;;OIKAWA Tadao2003～Kansai University Research Grants 200304-200503

PapersIn refereedAcademic JournalCo-authored2003～Kansai University Research Grants 200304-200503

PapersUnrefereedSingle-AuthorOIKAWA,Tadao2003～

Research reportUnrefereedOtherCo-authored2003～

PapersIn refereedAcademic JournalCo-authored;;Cytophage2002/12/20～

PapersThermostable aldehyde dehydrogenase from psychrophile, Cytophaga sp. KUC-1: enzymological characteristics and functional propertiesIn refereedAcademic JournalCo-authoredY. Yamanaka;T. kazuoka;M. Yoshida;K. Yamanaka;T. Oikawa;K. Sodapsychrophile;aldehyde dehydrogenase;Cytophaga;Thermostable enzymeBiochemical and Biophysical Research Communication2002/10～We found the occurrence of NAD(P)+ -dependent aldehyde dehydrogenase (EC1.2.1.5) in the cells of a psychrophile from Antarctic seawater, Cytophaga sp. KUC-1, and purified to homogeneity. About 50% of the enzyme activity remained even after heating at 50℃ for 65min and the highest activity was observed in the range of 55-60℃. The enzyme was thermostable and thermophilic, although it was derived from a psychrophile. The circular dichroism at 222nm of the enzyme showed a peak at 32℃. This temperature was closely similar to the transition temperature in the Arrhenius plots. The stereospecificity for the hydride transfer at C4-site of nicotinamide moiety of NADH was pro-R . The gene encoding the enzyme consisted of an open reading frame of 1506-bp encoding a protein of 501 amino acid residues. The significant sequence identity (61%) was found between the Cytophaga and the Pseudomonas aeruginosa enzymes, although their thermostabilities are completely different. Ⓒ2002 Elsevier Science (USA). All rights reserved.

PapersIn refereedAcademic JournalCo-authored2001/12/20～

PapersIn refereedAcademic JournalCo-authored2001/12/20～

PapersIncreased Transglycosylation Activity of Rhodotorula glutinis Endo-beta-Glucanase in MediaContaining Organic Solvent,In refereedCo-authoredEndo-β-Glucanase;Rhodotorula glutinis;TransglycosylationBiosci. Biotechnol.Biochem.Vol.65 pp.1889-18922001/5～The transglycosylation of p-nitrophenyl-β-D-cellotrioside to cellotetraose catalyzed by endo-1,4-β-glucanase (cellulase, EC 3.2.1.4) from a psychrotrophic yeast, Phodotorula glutinis KUJ 2731, was increased by addition of a miscible organic solvent in the reaction mixture.Among various organic solvents tested, acetone was most effective.The transglycosylation activity increased with an increase in acetone concentrations, while hydrolysis activity was suppressed .The transglycosylation preferably occurred at acidic pH with the optimum pH at 2 in 10mΜ Gly-HCl buffer. The optimum temperature of transglycosylation was found to be 50℃ in the presence of 40℃ acetone.

PapersOne-pot chemo-enzymatic enantiomerization of racemates ,In refereedCo-authoredChemo-Enzymatic enantiomerization;One-potJournal of Molecular Catalysis,Vol.11 pp . 149-1532001/4～A new one-pot chemo-enzymatic procedure was developed for enantiomerization of racemates based on enzymatic enantiospecific oxidation of a substrate and chemical non-enantiospecific reduction of the product. The principle is shown as follows for the L-proline production. L-Pro ←ゥゥゥゥゥゥゥゥゥゥゥゥゥ・　　　　　　　　　　　　　　　　　・　　　　　　　　　　　　　　　　　・　ゥゥゥゥゥゥゥゥゥゥゥゥゥゥゥゥ・ 　↓　　　　　　　　　　　　　　　・D-Pro―D-Amino acid oxidase→△1-Pyrroline-2-carboxylic acidL-Proline and L-pipecolate were produced from racemic proline and pipecolate by means of D-amino acid oxidase and sodium borohydride in high yield in this reaction system [J. W. Huh, K. Yokoigama, N. Esaki, K. Soda, Biosci., Biotechnol., Biochem. 56 (1992) 2081].DL-and L-Lactate were DL-enantiomerized in a one-pot reaction systems containing L-lactate oxidase and sodium borohydride in the similar mannar [S. Mukoyama, K. Yamanaka, T. Oikawa, K. Soda, Nippon Nogei Kagaku Kaishi 73 (1999) 62].Pyruvate was also converted to an equimolar amount of D-lactate in the same system. D-α-Hydroxybutyrate can be produced from the DL- and L-isomers, and α-ketobutyrate in the same manner though slowly.This method is applicable to production of other chiral compounds from the corresponding racemates. Ⓒ2001 Elsevier Science B. V. All right reserved.

PapersChemo-Enzymatic D-Enantiomerization of DL-Lactate,In refereedCo-authoredChemo-Enzymatic D-EnantiomerizationBiotechnology and Bioengineering,Vol . 73 pp . 80-822001/3～Abstract:┏B/┓ We investigated the total conversion of racemic lactate, L-lactate, and pyruvate into D-lactate, which is very useful as a starting material for the synthesis of chiral compounds and much more valuable than the L-enantiomer by means of coupling of L-specific oxidation of the recemate with L-lactate oxidase and non-enantiospecific reduction of pyruvate to DL-lactate with sodiumborohydride.In this one-pot system, L-lactate was enantiospecifically oxidized to an achiral product, pyruvate, which was chemically reduced to DL-lactate leading to a turnover.Consequently, either DL-lactate, L-lactate, or pyruvate was fully converted to the D-enantiomer.We optimized the reaction conditions: DL-lactate was converted to D-lactate in 99% of the theoretical yield and with more than 99% enantiomeric excess.DL-α-Hydroxybutyrate and α- ketobutyrate were converted also to D-α- hydroxybutyrate in the same way, though slowly.Ⓒ2001

PapersPsychrophilic valine dehydrogenase of the antarctic psychrophile, Cytophaga sp. KUC-1: purification,molecular characterization, and expression,In refereedCo-authoredprimary structure;Cytophaga;valine dehydrogenaseEuropean Journal of Biochemistryvol.268, pp4375-43832001/2～We found the occurrence of valine dehydrogenase in the cell extract of a psychrophilic bacterium, Cytophaga sp.KUC-1, isolated from Antarctic seawater and purified the enzyme to homogeneity.The molecular mass of the enzyme was determined to be ≈ 154 kDa by gel filtration and that of the subunit was 43 kDa by SDS/PAGE: the enzyme was a homotetramer.The enzyme required NAD+ as a coenzyme, and catalyzed the oxidative deamination of L-valine, L-isoleucine, L-leucine, and the reductive amination of α-ketoisovalerate, α-ketovalerate, α-ketoisocaproate, and α-ketocaproate.The reaction proceeds through an isoordered bi-bi mechanism.The enzyme was highly susceptible to heat treatment and the half-life at 45℃ was estimated to be 2.4 min.The kcat/Km(μ-1・-1)values for L-valine and NAD+ at 20℃ were 27.48 and 421.6, respectively.The enzyme showed pro-S stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of coenzyme. The gene encoding valine dehydrogenase was cloned into Escherichia coli(Novablue), and the primary structure of the enzyme was deduced on the basis of the nucleotide sequence of the gene encoding the enzyme.The enzyme contains 370 amino-acid residues, and is highly homologous with S.coelicolor ValDH(identity, 46.7%) and S.fradiaeValDH(43.1%).Cytophaga sp.KUC-1 ValDH contains much lower numbers of proline and arginine residues than those of other ValDHs.The changes probably lead to an increase in conformational flexibility of the Cytophaga enzyme molecule to enhance the catalytic activity at low temperatures.

PapersFragmentary Form of Thermostable Leucine Dehydrogenase of Bacillus stearothermophilus : ItsConstruction and Reconstitution of Active Fragmentary Enzyme ,In refereedCo-authoredamino acid dehydrogenase;fragmentary enzyme;Bacillu stearo-thermophilus;leudine dehydrogenaseBiochemical and Biophysical Research CommunicationsVol.280 pp . 1177-11822001/1～X-ray crystallographic studies revealed that various amino acid dehydrogenases fold into two domains in each subunit, a substrate-binding domain and an NAD(P)+-binding domain(Baker, P.J., Turnbull, A. P., Sedelnikova, S. E., Stillman , T. J., and Rice, D. W.(1995) Structure 3, 693-705).To elucidate the function and folding process of these two domains, we have genetically constructed a fragmentary form of thermostable leucine dehydrogenase of Bacillus stearothermophilus consisting of an N-terminal polypeptide fragment corresponding to the substrate-binding domain including an N-terminus, and a C-terminal fragment corresponding to the NAD+-binding domain.The two peptide fragments were expressed in separate host cells and purified. When both fragments were mixed, the leucine dehydrogenase activity with a specific activity of 1.4% of that of the wild-type enzyme appeared.This suggests that both peptide fragments mutually recognize each other, associated and fold correctly to be catalytically active, although the activity is low.However, the fragmentary form of enzyme produced catalyzed the oxidative deamination of L-leucine, L-isoleucine, and L-valine with broad substrate specificity compared to that of the wild-type enzyme.The fragmentary enzyme retained more than 75% of the initial activity after heating at 50℃ for 60 min.The fragmentary enzyme was more stable on heating than separate peptide fragments.These results suggest that the two domains of leucine dehydorogenase probably fold independently, and the two peptide fragments interact and associate with each other to form a function active site.

PapersRelationship between cytocidal activity and glutathione-S-transferase inhibition using doxorubicin coupled to stereoisomers of glutathione with different substrate specificityIn refereedAcademic JournalCo-authoredY.Hashizume;T.Asakura;T.Oikawa;T.Yamauchi;K.Soda;K.OhkawaStereoisomers of glutathione;conjugate;Glutathione-doxorubicin;Cytotoxicity;ApotosisAnti-Cancer Drug2001～To deterumine the cytotoxic mode of action of a glutathione (GSH)-doxorubicin(DXR)conjugate, which exhibited potent cytotoxicity against various multidrug-resistant as well as DXR-sensitive cell lines,the molecular interaction between covalent GSH-DXR conjugates and glutathione-Stransferase (GST), a possible molecular target of the conjugates, was investigated. The following four GSH molecules with stereoisomeric forms were prepared:L-Glu-L-Cys-Gly(LL-GSH),D-Glu-L-Cys-Gly(DD-GSH),L-Glu-D-Cys-Gly(LD-GSH) and D-Glu-D-Cys-Gly(DD-GSH). The enzymic activity of GST against each GSH stereoisomer was 88, 38, 8 and 4 nmol/mg/min, respectivery, syggesting that the L-form cysteine residue in the molecule was an important substrate of GST. Addition of DXR conjugated with each isomer(10M)to a GSH-containing GST assay mixture inhibited the GST activity to 32% for LL-GSH -DXR, 16% for DL-GSH-DXR and 61% for LD-GSH-DXR as compared with the solvent control. Moreover, IC50 values for these conjugates were 30, 20 and 250 nM, respectively. The cytocidal activity of each conjugate corresponded to the substrate specifity of GST activity for the GSH isomer.These conjugates bound to the GST molecule, and the binding ability was 0.746, 0.627 and 0.462 mol/mol of GST for LL-GSH-DXR, DL-GSH-DXR and LD-GSH-DXR, respectively. These findings suggested that GSH-DXR interacted with the substrate-binding site of the GST molecule and inhibition of GST activity exhibited potent cytotoxicity.

PapersUnrefereedSingle-AuthorOIKAWA Tadao2000～

PapersProduction of D-Glutamate from L-Glutamate with Glutamate Racemase and L-Glutamate oxidaseIn refereedAcademic JournalCo-authoredTadao Oikawa;Mayumi Watanabe;Hidemi makiura;Hitoshi Kusakabe;Kazuhiro Yamade.;Kenji SodaL-Glutamate oxidase;D-Glutamate productionBiosci. Biotech. Biochem.1999/12～We studied production of D-glutamate from L-glutamate using a bioreactor consiting of two columns of sequentially connected immobilized glutamate racemase (EC 5.1.1.3, from Bacillus subtilis IFO 3336) and L-glutamate oxidase (EC 1.4.3.11, from Streptomyces sp. X119-6: L-glutamate was racemized by the glutamate racemase column, and then L-glutamate was oxidized by the L-glutamate oxidase column. Consequently only D-glutamate remained, and was easily separated from the α-ketoglutarate formed by anion-exchange chromatograph. Both enzymes were highly stabilized by immobilization. The pH and temperature optima of immobilized glutamate racemase (pH 8, 40℃) were similar to those of immobilized L-glutamate oxidase (pH 7, 50℃). Accordingly, we connected the two columns tandemly to do both enzyme reactions under the same conditions. Actually 4.5 μ mol of D-glutamate was produced and isolated from 10 μ mol of L-glutamate, about 90% of the theoretical yield.

Academic presentationUltrathermostable Alanine Aminotrausterases of Hyperthermophilic Archaeron, Pyrococcus furiosusUnrefereedOtherCo-authoredKenji Soda;Satoru Sakaguchi;Tadao OikawaAlanine Aminotransferase;Ultrathermostable enzyme1999/10～We found the occurrence of alanine aminotransferase (AlaAT, EC 2.6.1.2) isozymesⅠand Ⅱin a hyperthermophilic archaeron, Pyrococcus furiosus DSM 3638 grown on pyruvate as a sole carbon source, and purified to characterize it. The enzyme was homotetrameric, and the molecular mass was about 168,000 Da. The enzyme showed the maximum activity at pH 9, and Km for L-alanine and 2-oxoglutarate were 2.3 and 2.4 mM, respectively. The enzyme catalyzes the pyridoxal phosphate-dependent transamination of alanine, 2-amino-butyrate, norvaline, and norleucine with α-ketoglutarate, whereas phenylalanine, tyrosine, and tryotophane are insert as an amino donor. The inhibition experiment showed that both pyridoxal phosphate and sulfhydryl group were directly involved in catalysis. The arrhenius plots of from P. furisus: activation energy, 57 kJ/mol. The consensus sequence (YAVR of alanine aminotransferase Ⅱwas found as 1XKASKRAMSIXYAIRNVVLP-in the N-terminal region.

Academic presentationBacterial Guanidino butyrase acting on D-ArginineUnrefereedOtherCo-authoredNoriaki Arakawa;Tadao Oikawa;kenji SodaGuanidino butyrase1999/10～We found the occurrence of D-arginase in cells of Acinetobacter haemolyticus KUJ 8661 grown in the L-arginine medium, purified and characterized the enzyme. The molecular weight and subunit of the enzyme was estimated to be about 232,000 and 40,000, respectively, suggesting that the enzyme is a homohexamer. The enzyme does not act on only D-arginine, but 4-guanidinobutrate, 3-guanidinopropionate and L-arginine. The Km values for 4-guanidinobutyrate and D-arginine were determined 3.9 and 22 mM, respectively. Accoringly, it is more pertinent to name the enzyme guanidinobutyrase (guanidinobutyrate amidinohydrorase, Ec.3.5.3.7). The optimum pH was 9.0. Incubation of the enzyme in part. The activity of the inactivated enzyme was substantially restored by incubation with Co2+.

PapersVitamin B6 enzymes participating in selenium amino acid metabolismIn refereedAcademic JournalCo-authoredKenji Soda;Tadao Oikawa;Nobuyoshi EsakiSelenium amino acid;Vitamin B6 enzymeBiofactors1999/6～Various vitamin B6 enzymes play important roles in mammalian and microbial metabolism of selenium amino acids. Selenocysteine is synthesized from selenohomocysteine by catalysis of cystathionine β-synthase and cystathionine γ-lyase, which both require pyridoxal phosphate. Selenocysteine β-lyase, a new B6-enzyme, exclusively catalyzes β-elimination of selenocysteine, and occurs in mammalian systems and bacteria. Methionine γ-lyase, cysteine desulfurase, cysteline sulfinate desulfinase, and D-selenocystine α,β-lyase, whice are B6-enzymes, act on cysteine, cysteine sulfinate, D-cystine, and their derivatives, and their selenium conuterparts indiscriminately. Their reaction mechanisms are comparatively described.

PapersStereoisomers of Glutathione : Preparation and Enzymatic ReactivitiesIn refereedAcademic JournalCo-authoredTadao Oikawa;Takahiro Yamauchi;Hidehiko Kumagai;Kenji SodaGlutatione;StereoisomerJ Nutr Sci Vitaminol1999/4～We synthesized a series of stereoisomers of glutathione (GHS) and glutathione disulfide (GSSG) by the solid-phase method. These peptides were used to examine their reactivities with enzymes acting on glutathione. The glutathione reductase of yeast acted only on LL-GSSG. Glutathione S-transferase catalyzed the conjugation of 1-chloro-2,4-dinitrobenzene sith LL-GSH and DL-GSH (Km(mM): for LL-GSH, 0.035; and for DL-GSH, 0.62), but the DD- and LD-diastereomers were inert. γ-Glutamyl transpeptidase catalyzed the transfer of γ-glutamyl moiety of LL-GSH and DL-GSH to taurine foming γ-glutamyl taurine and cysteinyl taurine (Km(mM): for LL-GSH, 0.336; and for DL-GSH, 0.628), but the other diastereomers were not the substrates. The occurrence of L-cysteinyl residue in the tripeptides is required for the glutathione analogue to be a substrate of the enzymes.

Academic presentationAlanine Aminotransferase Ⅱ from Hyperthermophilic Archaeron, Pyrococcus furiosus : Primary structureUnrefereedOtherCo-authoredSatoru Sakaguchi;Tadao Oikawa;Shun-ichi Kuroda;Katsuyuki Tanizawa;Kenji SodaAlanine aminotransferase1999～Alanine aminotransferase isozyme Ⅱ (AlaAT Ⅱ) purified from Pyrococcus furiosus DSM 3638 was digested with lysylendopeptidase, and the peptides were purified by reversed phase high performance liquid chromatography. The amino acid sequences of the peptides were analyzed by protein sequencer. N-Terminal and internal amino acid sequences of the enzyme were highly homologous to those of thermostable aspartate aminotransferase of P. horikoshii OT3. Based on these peptides sequences, the primers for polymerase chain reaction (PCR) were designed and AlaAT Ⅱ gene in chromosomal DNA of P. furiosus DSM 3638 was amplified by PCR. The DNA purified from agarose gel was ligated with pT7 Blue T-vector and tansformed into NovaBlue. The plasmid prepared by alkali miniprep method was purified by polyethylene glycol, and the insert (1.0 kb) was analyzed by DNA sequencer. The partial primary structure of AlaAT Ⅱ determined by DNA sequence agreed with the internal amino acid sequences, and the total DNA of AlaAT Ⅱ was obtained by genome-walking PCR.

Academic presentationUltrathermostable Alanine Aminotransferase Ⅱ of Hyperthermophilic Archaeron, Pyrococcus furiosus : Purification and characterizationUnrefereedOtherCo-authoredTadao Oikawa;Satoru Sakaguchi;Kenji SodaArchaeron;Alanine aminotransferase1999～Alanine aminotransferase (EC 2.6.1.2) isozyme Ⅱ (AlaAT Ⅱ) was purified to homogeneity from hyperthermophilic archaeron Pyrococcus furiosus DSM 3638 grown on pyruvate as a sole carbon source. The enzyme was homotetrameric, and the molecular mass was about 168,000 Da. The enzyme showed the maximum activity at pH 9, and K m for L-alanine and 2-oxoglutarate were 2.3 and 2.4 mM, respectively. The enzyme catalyzes the pyridoxal 5'-phosphate-dependent transamination of L-alanine, 2-amino-L-butyric acid, L-norvaline, and L-norleucine with α-ketoglutarate, whereas L-phenylalanine, L-tyrosine, and L-tryotophane are inert as an amino donor. The inhibition experiment showed that both pyridoxal 5'-phosphate and sulfhydryl groups were directly involved in catalysis. The arrhenius plots of the enzyme showed a single phase not similar to those of other enzymes from P. furiosus: activation energy, 57 kJ/mol. The consensus sequence (YAVR) of alanine aminotransferase was found as 1XKASKRAMSIXYAIRNVVLP-in N-terminal region.

PapersEndo-beta-Glucanase Secreted by a Psychrotrophic Yeast : Purification and CharacterizationIn refereedAcademic JournalCo-authoredTadao Oikawa;Yasuyuki Tsukagawa;Kenji SodaPsychrotrophic Yeast;Endo-β-GlucanasBiosci. Biotech. Biochem1998/9～A psychrotrophic yeast, Rhodotorula glutinis KUJ 2731, isolated from soil, effectively produced an extracellular endo-β-glucanase (EC 3.2.1.4). The enzyme was monomeric, and the molecular mass was about 40,000 Da. The N -terminal amino acid sequence was H-Ser-Leu-Pro-Lys-Leu-Gly-Gly-Val-Asp-Leu-Ala-Gly-Leu-Asp-Ile-Gly-Lys-Asp-Lys-Asn-. α-Helix content was calculated to be about 32.6%. The isoelectric point was 8.57. The activation energy was 20.9 kJ/mol, which was much smaller than that of mesophilic enzymes. The enzyme was active at temperatures from 0 to 70℃, sith a highest initial velocity at 50℃ similar to other psychrotrophic enzymes. The enzyme was inhibited by Hg2+. The enzyme catalyzed hydrolysis of carboxymethyl cellulose with an apparent Km of 1.1% and V max of 556 μ mol/min mg. Products from the enzymatic hydrolysis of carboxymethyl cellulose by the enzyme were glucose, cellobiose, and cellotriose. The enzyme also catalyzed the transglycosylation of p -nitrophenyl-β-cellotrio-side to cellotetraose.

CommentaryThermotorelant D-Amino acid transferase:Characterization and Reaction MechanismUnrefereedIn-house publicationCo-authoredKenji Soda;Tadao Oikawa;Nobuyoshi Esaki;Tohru Yoshimura1998/3～

CommentaryTrace elements and Euzymes:Reaction Mechanism of Enzyme ActivationUnrefereedIn-house publicationCo-authoredNobuyoshi Esaki;Tadao Oikawa;Kenji Soda1998/1～

BookCo-authored chapterOIKAWA,Tadao1998～

Research reportIn refereedOtherSingle-AuthorOIKAWA Tadao1998～

PapersA Novel Type of D-Mannitol Dehydrogenase from Acetobacter xylinum : Occurrence, Purification, and Basic PropertiesIn refereedAcademic JournalCo-authoredTadao Oikawa;Junji Nakai;Yasuyuki TsukagawaAcetobacter xylinum;Mannitol DehydrogenaseBiosci. Biotech. Biochem.1997/10～We purified a novel type of D-mannitol dehydrogenase, which contains a c -type cytochrome and an unknown chromophore in the soluble fraction of an acetic aced bacterium, Acetobacter xylinum KU-1, to homogeneity. The enzyme showed the maximum activity at pH 5 and 40℃. It was stable up to 60℃ at pH 6, and was inhibited by Hg2+ and p-quinone (KI=0.18mM). The molecular weight of the enzyme was about 140,000, and those of the subunits were 69,000 51,000, and 20,000; the enzyme is hetero-trimeric and contained 8 g-atoms of Fe per mole. The α-helix content was estimated to be about 52.9%. The enzyme catalyzed phenazine methosulfate dependent oxidation of D-mannitol with an apparent Km of 98 μ M (for D-mannitol) and Vmax of 213μ mol/min/mg. The reduced form of the enzyme showed the absorption maxima at 386, 416, 480, 518, 550, and 586 nm, which are attributable to a c-type cytochrome in the enzyme.

PapersEndo-beta-Glucanase from Acetobacter xylinum KU-1 : Purification and CharacterizationIn refereedAcademic JournalCo-authoredTadao Oikawa;Toshinori Kamatani;Takeshi Kaimura;Minoru Ameyama;Kenji SodaAcetobacter xylinum;Endo-β-GlucanaseCurrent Microbiology1997/5～A cellulose-producing acetic acid bacterium, Acetobacter xylinum KU-1, abundantly produces an extracellular endo-β-glucanase (EC 3.2.1.4) in the culture broth. The enzyme was purified to homogeneity by DEAE- and CM- Toyopearl 650M ion-exchange chromatography, Butyl-Toyopearl 650M hydrophobic chromatography, and Toyopearl HW-50 gel filtration. The purified enzyme showed the maximum activity at pH 5 and 50℃: it was stable up to 50℃ at pH 5, activated by Co2+, and competitively inhibited by Hg2+; the apparent Ki was 7 μ M. The molecular weight of the enzyme was determined to be about 39,000 by sodium dodesyl sulfate/polyacrylamide ge electrophoresis, and about 41,000 by Toyopearl HW-50 gel filtration; the enzyme is monomeric. The enzyme hydrolyzed carboxymethylcellulose with an apparent Km of 30 mg/ml and Vmax of 1.2 μ M/min. It hydrolyzed cellohexaose to cellobiose, cellotriose and cellotetraose, and also cellopentaose to cellobiose and cellotriose, but did not act on cellobiose, cellotriose, or cellotetraose.

PapersUnrefereedSingle-AuthorOIKAWA,Tadao1997～

PapersUnrefereedCo-authoredOIKAWA Tadao1996～

PapersProduction of Cellulose from D-Arabitol by Acetobacter xylinum KU-1In refereedAcademic JournalCo-authoredTadao Oikawa;Takafumi Morino;Minoru AmeyamaAcetobacter xylinum;Arabitol;Cellulose productionBiosci. Biotech. Biochem1995/8～We found that Acetobacter xylinum KU-1 produced cellulose from D-arabitol. The maximum cellulose production was obtained when it was grown in a medium containing 2.0% D-arabitol, 1.0% tryptone, and 1.0% yeast extract (pH 5) at 30℃ for 96h statically. The productivity was more than 6 times as much as that of D-glucose[productivity (mg/ml-medium): from D-arabitol, 12.4; from D-glucose, 2.0].

PapersProduction of Cellulose from D-Mannitol by Acetobacter xylinum KU-1In refereedAcademic JournalCo-authoredTadao Oikawa;Toshiyuki Ohtori;Minoru AmeyamaAcetobacter xylinum;Mannitol;Cellulose productionBiosci. Biotech. Biochem1995/2～We found that Acetobacter xylinum KU-1 produced cellulose from D-mannitol. The optimum culture conditions for cellulose production were 1.5% D-mannitol, 0.5% Polypeptone, 2.0% yeast extract, pH 5, 30℃, and 48h. The amount of cellulose from D-mannitol was more than 3 times as much as that from D-glucose under the same culture conditions [productivity (mg/ml-medium): from D-mannitol, 4.6; from D-glucose, 1.2].

PapersDetection of Carboxymethyl Cellulase Activity in Acetobacter xylinum KU-1In refereedAcademic JournalCo-authoredTadao Oikawa;Miho takagi;Minoru AmeyamaAcetobacter xylinum;Carboxymethyl CellulaseBiosci. Biotech. Biochem.1994/11～We detected carboxymethyl cellulase activity in a crude extract of Acetobacter xylinum KU-1. The enzyme activity was detected when glycerol, D-fructose, D-mannitol, D-glucose, D-arabitol, D-sorbitol, or carboxymethyl cellulose was used as a carbon source. The optimum pH was found to be 4.0, while the optimum temperature was 50℃. The enzyme activity was inhibited characteristically by the addition of Hg2+

PapersDetection of Dye-linked D-Mannitol Dehydrogenase Activity in Acetobacter xylinum KU-1In refereedAcademic JournalCo-authoredTadao Oikawa;Minoru AmeyamaAcetobacter;Mannitol dehydrogenaseBiosci. Biotech. Biochem1993/9～We detected dye-linked D-mannitol dehydrogenase activity in the crude extract of Aectobacter xylinum KU-1. The enzyme activity was specific for D-mannitol, and not pyridine nucleotide (NAD+, NADP+)-dependent. The optimal pH was found to be 5.0, while the optimal temperature was at 50℃. The enzyme activity was inhibited by p-quinone noncompetitively.Kansai University Research Grants 199204-199303

PapersSynthesis and Characterization of the Selenium Analog of Glutathione DisulfideIn refereedAcademic JournalCo-authoredTakashi Tamura;Tadao Oikawa;Akira Ohtaka;Nobutaka Fujii;Nobuyoshi Esaki;Kenji SodaGlutathione;Selenium analogAnal. Biochem1993/1～We synthesized the selenium analog of glutathione disulfide by a liquid phase method and named it glutaselenone (I.e., γ-L-glutamyl-L-selenocysteinylglycine) diselenide. The selenol of selenocysteine was protected by the p-methoxybenzyl group, which was removed by acidolysis with trifluoroacetic acid in the presence of thioanisol. The overall yield of the final product, glutaselenone diselenide, was about 9% based on the starting compound, Se-(p-methoxybenzyl)-L-selenocysteine. Glutaselenone diselenide showed a broad absorption band between 270 and 400 nm and circular dichroism bands around 270 nm (positive) and 330 nm (negative), which were attributable to diselenide bond.Grant-in-Aid for Scientific Research 199204-199304

BookCo-authored chapterOIKAWA Tadao1993～

PapersA Selenium Analogue of Metallothionein Chemical Synthesis and CharacterizationUnrefereedCo-authoredOIKAWA TadaoToyobo Biotechnology Fundation Record of Activitiespp. 196-1971993～

PapersUnrefereedCo-authored1992～

PapersUnrefereedCo-authored1992～

PapersMetalloselenonein, the Selenium Analogue of Metallothionein : Synthesis and Characterization of its complex with Copper IonsIn refereedAcademic JournalCo-authoredTadao Oikawa;Nobuyoshi Esaki;Hidehiko Tanaka;Kenji SodaMetallothionein;MetalloselenoneinProc. Natl. Acad. Sci. USA1991/4～We used an automated peptide synthesizer to produce a peptide, metalloselenonein, that contains selenocysteine residues substituted for all cysteine residues in Neurospora crassa copper metallothionein. Metalloselenonein binds 3 mol of Cu(I) per mol. This adduct shows a broad absorption band between 230 and 400 nm and a fluorescence band at 395 nm, which can be attributed to copper-selenolate coordination. The circular dichroism spectrum of the copper-metalloselenonein complex shows a positive band around 245 nm attributable to asymmetry in metal coordination.Grant-in-Aid for Scientific Research 199004-199103

PapersPurification and Properties of Dye-linked Aldehyde Dehydrogenase in Rhodopseudomonas acidophila M402In refereedAcademic JournalCo-authoredKei Yamanaka;Hideaki Iino;Tadao OikawaRhodopseudomonas acidophila;Aldehyde dehydrogenaseAgric. Biol. Chem1991/4～Rhodopseudomonas acidophila M402 grown under aerobic-dark condition shows two bands of dye-linked aldehyde dehydrogenase activity by polyacrylamide gel electrophoresis. The slower migrating band coincided with the dye-linked aromatic alcohol deyhdrogenase that was active on aromatic and aliphatic alcohols and aldehydes. The faster-migrating band was a new dye-linked aldehyde dehydrogenase. The latter enzyme was purified 125-fold by ultracentrifugation and column chromatographies on DEAE-cellulose, Bio GelHTP, and Sepharose CL-6B. The enzyme has a relative molecular mass of 70,000 daltons, and a subunit size of 35,000 dalton indicates the dehydrogenase has a dimeric structure. The isoelectric point was pH 4.74. NAD+ and NADP+ do not act as electron acceptors. The enzyme was active specifically on straight chain aldehydes (C3-C10) rather than on benzaldehyde and its substitutes. Aromatic and aliphatic alcohols were inert for this enzyme. The Michaelis constant (mM) were 1.6, 0.6, 0.9, 3.6, 0.5, 0.3, 0.1, 0.9, 0.6, 0.3, and 0.1 for benzaldehyde, m -hydroxybenzaldehyde, m -anisaldehyde, vanillin, propionaldehyde, butyraldehyde, hexaldehyde, heptaldehyde, octaldehyde, nonaldehyde, and decaldehyde, respectively.

PapersIn refereedCo-authored1991～

PapersUnrefereedCo-authoredOIKAWA,Tadao1991～

PapersIn refereedCo-authored1990～

PapersChemical Synthesis and Expression of Copper Metallothionein Gene of Neurospora crassaIn refereedAcademic JournalCo-authoredManabu Sugimoto;Tadao Oikawa;Nobuyoshi Esaki;Hidehiko Tanaka;Kenji SodaNeurospora crassa;MetallothioneinJ. Biochem.1988/7～The gene coding for the Neurospora crassa copper metallothionein (MT) was synthesized and inserted in the lacZ' gene of pUC18 plasmid to give the same translational reading frame as the latter gene. The MT-β-galactosidase fused gene was expressed in Escherichia coli to produce a fused protein in which the amino and carboxy termini of MT are linked to the β-galactosidase through methionine residues. An MT derivative containing an extra homoserine residue at the carboxy terminus was prepared by cyanogen bromide cleavage of the fused protein followed bya a reverse-phase HPLC separation. The spectral features of the MT derivative and its copper complex were similar to those of the corresponding native MTs.

PapersSynthesis of Biologically Active Selenium-containing Amino Acids and PeptidesIn refereedAcademic JournalCo-authoredHidehiko Tanaka;Nobuyoshi Esaki;Manabu Sugimoto;Tadao Oikawa;Patrick Chocat;Kenji Sodapeptide;aminoacid;SeleniumPhosphorus and Sulfur1988/7～We here describe the synthesis of selenium amino acids with O-acetylhomoserine sulfhydrylase, partially purified from baker's yeast. The enzyme was found to catalyze the syntehses of L-selenocystine and L-selenohomocystine from sodium diselenide with the corresponding acety1-derivatives of serine and homoserine, respectively. L-serine O-sulfate also serves as a substrate of the β-replacement reaction. Sodium diselenide is less efficient as a substituent donor than the physiological substrate, sodium sulfide and inhibits the enzyme at high concentrations. Therefore, limited amounts of sodium diselenide were added to the reaction mixture to increase the yield (about 60%). This provides a facile method to produce optically active selenocystine and selenohomocystine. In addition, we developed a convenient method for the syntehsis of a new seleniumcontaining amino acid, L-selenodjenkolic acid (3,3 -methyl- enediselenobis(2-amino-prpionic acid)) from L-selenocystine thus prepared. This amino acid undergoed α,β-elimination to produce pyruvate, formaldehyde, ammonia and selenium by bacterial methionine γ-lyase under aerobic conditions.

PapersIn refereedCo-authored1988～

PapersUnrefereedSingle-Author1988～

Research Activities Overseas

• Kansai University's Overseas Research Program(long term)Aug. 2001-Aug. 0,Germany Forsehungszentrium JUlich, IBT-1

Participation in International Conferences

• Dutch-Japanese Workshop on Biocatalysis Sep.2003
• Ⅴｔｈ International Symposium on the Chemistry of Selenium and Tellurium 1987
• Ⅳth International Symposium of Selenium Biology and Medicine 1988
• 1st Swiss-Japanese Joint Meeting on Bioprocess Department 1988
• Second Meeting of the International Society for trace element Research in Humans 1989
• Chemical Synthesis and Characterization: The 1989 International Chemical Congress of Pacific Basin Societies 1989
• The Ⅵth International Conference on the Chemistry of Selenium and Tellurium 1991
• The Ⅵth International Conference on the Chemistry of Selenium and Tellurium 1991
• The First International Congress on "Vitamines and Biofactors in Life Science" 1991
• 3 rd International Congress on Vitamins and Related Biofactors 1998
• 3 rd International Congress on Vitamins and Related Biofactors 1998
• Purification and Characterization: 10th International Symposium on Vitamin B6 and Carbonyl Catalysis and 4th Meeting on PQQ and Quinoproteins 1999
• Primary Structure: 10th International Symposium on Vitamin B6 and Carbonyl Catalysis and 4th Meeting on PQQ and Quinoproteins 1999
• Purification, Characterization, and Structure: 6th International Union of Biochemistry and Molecular Biology Seoul Conference 1999
• 6th International Union of Biochemistry and Molecular Biology Seoul Conference 1999
• 6th International Union of Biochemistry and Molecular Biology Seoul Conference 1999
• Purification, Characterization, and Cloning: The 2000 International Chemical Congress of Pacific Basin Societies 2000
• Cloning and Primary Structure: The 2000 International Chemical Congress of Pacific Basin Societies 2000
• The 2000 International Chemical Congress of Pacific Basin Societies 2000
• The 2000 International Chemical Congress of Pacific Basin Societies 2000
• Cloning and Primary Structure: The 2000 International Chemical Congress of Pacific Basin Societies 2000
• The 11th German-Japanese Workshop on Enzyme Technology 2001
• The 2nd China-Japan International Conference on Vitamins 2001
• Japan-Italy Symposium New Trends in Enzyme Science and Technology 2001
• The 2nd International Symposium on Peptide and Protein Materials 2002
• 3rd International Symposium on Vitamin B6 2002
• 3rd International Symposium on Vitamin B6, PQQ, Carbonyl Catalysis and Quinoproteins
• 3rd International Symposium on Vitamin B6, PQQ, Carbonyl Catalysis and Quinoproteins 2002
• VAAM-Jahrestagung 2002
• An International Symposium held under auspices of the European Society for Marine Biotechnolog Greifswald 2002
• Geongju 2002
• Dutch-Japanese Workshop on Biocatalysis 2003
• The 12th Japanese-German Workshop on Enzyme Technology (Nov. 17-18, Ohtsu, Japan). 2003
• Italy-Japan Symposium 2003
• The 3rd China-Japan International Conference on Vitamins May15,2004-May 18,2004
• Japan‐Finland Joint Seminar on New Aspects in Microbial Biotechnology Oct.2,2004
• Tagungsband zum 2. Gemeinsamen Kongress der DGHM und VAAM Sep.26,2005
• 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress Jun.19,2006
• International Interdisciplinary Conference on Vitamins， Coenzymes， and Biofactors　2005 Nov.10,2005

Courses Taught

• Freshmen Seminar
• Biochemistry I
• Enzyme Chemistry
• Protein Engineering
• Exercises in Thesis Projects I and II
• Thesis Projects I
• Thesis Projects II
• SeminarI（Biotechnology)
• SeminarII（Biotechnology)
• SeminarIII（Biotechnology)
• SeminarIV（Biotechnology)
• Advanced Enzyme Chemistry
• Advanced Enzyme Chemistry