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NEW APPROACH TO SUSTAINABLE CHEMICAL RESOURCES BASED ON SEARCH, PRECISE ANALYSIS, AND APPLICATION OF BACTERIAL SUGAR-CARBOCYCLIZATION ENZYMES

Research Project

Project/Area Number 11356004
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Bioproduction chemistry/Bioorganic chemistry
Research InstitutionTOKYO INSTITUTE OF TECHNOLOGY

Principal Investigator

KAKINUMA Katsumi  TOKYO INSTITUTE OF TECHNOLOGY, GRADUATE SCHOOL OF SCIENCE AND ENGINEERING, PROFESSOR, 大学院・理工学研究科, 教授 (90092543)

Project Period (FY) 1999 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥37,860,000 (Direct Cost: ¥33,600,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2002: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2001: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2000: ¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1999: ¥12,300,000 (Direct Cost: ¥12,300,000)
Keywordssugar-carbocyclization enzymes / aminoglycoside antibiotics / metabolic engineering / 2-deoxy-scyllo-inosose / mechanism-based enzyme inhibitor / benzenoid resources / carbaglucose / secondary metabolism enzymes / 糖質還化酵素 / カナマイシン / アミノ基転移酵素 / 基質認識 / 部位特異的変異 / 糖質環化 / スーデオキシ-scyllo-イノソース合成酵素 / 反応機構 / カテコール / 持続可能資源 / バイオマス
Research Abstract

New paradigm for manufacturing sustainable chemical resources based on the CO_2 fixation is highly desirable to conserve fossil resources and to protect our living environments. An approach using carbohydrate-cyclizing enzyme, 2-deoxy-scyllo-inosose (DOI) synthase (DOIS), being originally involved in the biosynthesis of butirosins in Bacillus circulans, has been developed, and detailed analysis and alternative application of DOIS have been exploited. (1) DOIS of B. circulans was purified, and the responsible gene (btrC) was identified and expressed in E. coli. (2) Fermentation conditions of the transformed E. coli and preparative protocol of DOI were established. (3) Conversion of DOI into catechol or 1, 2, 4-triacetoxybenzene was confirmed by simple chemical treatment. (4) Detailed interaction between the enzyme and substrate was analyzed using substrates analogues to elucidate importance of specific hydrogen-bondings. (3) Active site mapping was pursued with mechanism-based inhibitor … More with aids of enzymatic digestion and LC/ESI/MS/MS analysis. (4) For alternative preparation of DOI by metabolic engineering, the biosynthetic gene cluster of butirosin was extensively analyzed. (5) A new transamination enzyme (BtrS) was identified to be responsible for the step next to DOIS, and was over-expressed in E. coli to confirm the function. (6) The btrS gene in B. circulans was disrupted by in frame deletion. Potential of the disruptant for DOI production is currently under investigation. (7) The over-expressed BtrS was utilized to prepare.2-deoxy-scyllo-inosamine as a potential resource for the development of pharmaceuticals. (8) DOI was successfully converted into carbaglucose which is a promising substitute for glucose without dietary load. (8) A new DOIS was identified from a Streptomyces sp. and overexpressed. Its function was confirmed. All these works appear to constitute significant fundamentals for the precise understanding of DOISs and for its application to the preparation of organic resources. Less

Report

(5 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Fumitaka Kudo: "Purification and Characterization of 2-Deoxy-scyllo-inosose Synthase Derived from Bacillus circulans. A Crucial Carbocyclization Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics"Journal of Antibiotics. 52[2]. 81-88 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Fumitaka Kudo: "Molecular Cloning of the Gene for the Key Carbocycle-forming Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminocyclitol Antibiotics and Its Comparison with Dehydroquinate Synthase"Journal of Antibiotics. 52[6]. 559-571 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Katsumi Kakinuma: "An Expeditious Chemo-Enzymatic Route from Glucose to Catechol by the Use of 2-Deoxy-scyllo-inosose Synthase"Tetrahedron Letters. 41[12]. 1935-1938 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yasumasa Ota: "Butirosin Biosynthetic Gene Cluster from Bacillus circulans"Journal of Antibiotics. 53[10]. 1158-1167 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tadashi Eguchi: "Importance of Specific Hydrogen-Bonding Donor-Acceptor Interactions for the Key Carbocycle-forming Reaction Catalyzed by 2-Deoxy-scyllo-inosose Synthase in the Biosynthesis of 2-Deoxystreptamine-containing Aminocyclitol Antibiotics"Journal of Organic Chemistry. 67[12]. 3979-3984 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hideyuki Tamegai: "Significance of the 20k-Da Subunit of Heterodimeric 2-Deoxy-scyllo-inosose Synthase for the Biosynthesis of Butirosin Antibiotics in Bacillus circulans"Bioscience, Biotechnology and Biochemistry. 66[7]. 1538-1545 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hideyuki Tamegai: "Identification of L-Glutamine:2-Deoxy-scyllo-inosose Aminotransferase Required for the Biosynthesis of Butirosin in Bacillus circulans"Journal of Antibiotics. 55[8]. 707-714 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Hideyuki Tamegai: "First Identification of Streptomyces genes involved in the Biosynthesis of 2-Deoxy-streptamine-containing Aminoglycoside Antibiotics. Genetic and Evolutionary Analysis of L-Glutamine:2-deoxy-scyllo-inosose Aminotransferase Genes"Journal of Antibiotics. 55[11]. 1016-1018 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Eriko Nango: "Reaction Stereochemistry of 2-Deoxy-scyllo-inosose Synthase, the Key Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics"Chemistry Letters. 32[5]. 438-439 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takashi Yukita: "Biosynthesis of (2R)-4-Amino-2-hydroxybutyric Acid, Unique and Biologically Significant Substituent in Butirosin"Journal of Antibiotics. 56[5]. 497-500 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Katsumi Kakinuma: "My Favorite Organic Synthesis"The Society of Synthetic Organic Chemistry, Japan. 58-59 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] F. Kudo, et al: "Purification and Characterization of 2-Deoxy-scyllo-inosose Derived from Bacillus circulans. A Crucial Carbo-cyclization Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics"J. Antibiot.. 52 [2]. 81-88 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] F. Kudo, et al: "Molecular Cloning of the Gene for the Key Carbocycle-forming Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminocyclitol Antibiotics and Its Comparison with Dehydroquinate Synthase"J. Antibiot.. 52 [6]. 559-571 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Kakinuma, et al: "An Expeditious Chemo-Enzymatic Route from Glucose to Catechol by the Use of 2-Deoxy-scyllo-inosose Synthase"Tetrahedron Lett.. 41 [12]. 1935-1938 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y. Ota, et al: "Butirosin Biosynthetic Gene Cluster from Bacillus circulans"J. Antibiot.. 53 [10]. 1158-1167 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Eguchi, et al: "Importance of Specific Hydrogen-Bonding Donor-Acceptor Interaction for the Key Carbocycle-forming Reaction Catalyzed by 2-Deoxy-scyllo-inosose Synthase in the Biosynthesis of 2-Deoxystreptamine- containing Aminocyclitol Antibiotics"J. Org. Chem.. 67(12). 3979-3984 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H. Tamegai, et al: "Significance of the 20k-Da Subunit of Heterodimeric 2-Deoxy-scyllo-inosose Synthase for the Biosynthesis of Butirosin Antibiotics in Bacillus circulans"Biosci. Biotechnol. Biochem.. 66 [7]. 1538-1545 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H. Tamegai, et al: "Identification of L-Glutamine : 2-Deoxy-scyllo-inosose Amino-transferase Required for the Biosynthesis of Butirosin in Bacillus circulans"J. Antibiot.. 55 [8]. 707-714 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H. Tamegai, et al: "First Identification of Streptomyces genes involved in the Biosynthesis of 2-Deoxy-streptamine-containing Amino-glycoside Antibiotics Genetic and Evolutionary Analysis of L-Glutamine : 2-deoxy-scyllo-inosose Aminotransferase Genes"J. Antibiot.. 55 [11]. 1016-1018 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] E. Nango, et al: "Reaction Stereochemistry of 2-Deoxy-scyllo-inosose Synthase The Key Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics"Chem. Lett.. 32 [5]. 438-439 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Yukita, et al: "Biosynthesis of (2R)-4-Amino-2-hydroxybutyric Acid, Unique and Biologically Significant Substituent in Butirosin"J. Antibiot.. 56 [6]. 497-500 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Kakinuma, et al, Tatsuta, K. Ed.: "D-Glucose : Extremely Versatile Resource for Organic Synthesis "My Favorite Organic Synthesis", Commemorative Publication of the 60th Anniversary of the Society of Synthetic Organic Chemistry, Japan"The Society of Synthetic Organic Chemistry, Japan. 58-59 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tadashi Eguchi: "Importance of Specific Hydrogen-Bonding Donor-Acceptor Interactions for the Key Carbocycle-forming Reaction Catalyzed by 2-Deoxy-scyllo-inosose Synthase in the Biosynthesis of 2-Deoxystreptamine-containine Aminocyclitol Antibiotics"Journal of Organic Chemistry. 67[12]. 3979-3984 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hideyuki Tamegai: "Significance of the 20k-Da Subunit of Heterodimeric 2-Deoxy-scyllo-inosose Synthase for the Biosynthesis of Butirosin Antibiotics in Bacillus circulans"Bioscience, Biotechnology and Biochemistry. 66[7]. 1538-1545 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hideyuki Tamegai: "Identification of L-Glutamine : 2-Deoxy-scyllo-inosose Aminotransferase Required for the Biosynthesis of Butirosin in Bacillus circulans"Journal of Antibiotics. 55[8]. 707-714 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hideyuki Tamegai: "First Identification of Streptomyces genes involved in the Biosynthesis of 2-Deoxy-streptamine-containing Aminoglycoside Antibiotics. Genetic and Evolutionary Analysis of L-Glutamine : 2-deoxy-syllo-inosose Aminotransferase Genes"Journal of Antibiotics. 55[1]. 1016-1018 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Hideyuki Tamegai: "Regulation of the Expression of Butirosin Biosynthetic Gene Cluster and Excretion System of Butirosin in Bacillus circulans"Journal of Antibiotics. 56(印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Eriko Nango: "Reaction Stereochemistry of 2-Deoxy-scvllo-inosose Synthase. The Key Enzyme in the Biosynthesis of 2-Deoxystreptamine-containing Aminoglycoside Antibiotics"Chemistry Letters. 32(印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Shohei Sakuda: "Biosynthetic Studies on the Cyclopentane Ring Formation of Allosamizoline an Aminocyclitol component of the Chitinase Inhibitor Allosamidin"Jornal of Organic Chemistry. 66-10. 3356-3361 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hideyuki Tamegai: "Significance of the 20KDa Subunit of Heterodimeric 2-Deoxy-Slyllo-inosose Synihase for the Biosynthesis Butirosin Antibiotics in Bacillus circulans"Bioscience, Biotechnology and Biochemistry. 66(印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Katsumi Kakinuma: "An Expedition Chemo-Enzymetic Route form Glucose to Catechol by the Uose of 2-Deoxy-scyllo-inosose Synthase"Tetrahedron Lett.,. 41・[12]. 1935-1938 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yasumasa Ota: "Batirosin Biosynthetic Gene Cluster from Bacillus circulans"J Antibiotics. 53・[10]. 1158-1167 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 柿沼勝己: "バイオマスを化学資源に-二次代謝酵素の可能性-"バイオサイエンスとインダストリー. 58・[12]. 39-40 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Fumitaka Kudo: "Molecular cloning of the gene for the key carbocycle-forming Enzyme in the Biosynthesis of 2-Deoxystreptamine Containing Aminocyclitol antibiotics and its Comparison with Dehydroquinate Synthase"J. Antibiotics. 52・3. 559-571 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Katsumi Kakinuma: "An Expeditions Chemo-Enzymatic Route from Glucose to Catechol by the Use of 2-Deoxy-scyllo -inosose Synthase"Tetrahedron Lett.,. 41・(印刷中). (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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