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Analysis of structure-function relationship in Fructosyltransferase of Zymomonas mobilis and its enzymatic production of useful saccharides

Research Project

Project/Area Number 07650959
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 生物・生体工学
Research InstitutionTottori University

Principal Investigator

YANASE Hideshi  Tottori University, Faculty of Engineering, Associate professor, 工学部, 助教授 (20158033)

Co-Investigator(Kenkyū-buntansha) KITA Keiko  Tottori University, Faculty of Engineering, Associate professor, 工学部, 助教授 (70234226)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsBiopolymer / Fructosyltransferase / Levan / Oligosaccharide / Random mutation / Site-directed mutagenesis / Glycosyltransferase / Zymomonas mobilis / NADP結合型酸化・還元酵素 / Zymomonas属細菌 / 糖アルコール / NAD(H)結合型酸化還元酵素
Research Abstract

Zymomonas mobilis produces an exocellular fructosyltransferase (EC2.4.1.10, sucrose : 2,6-beta-D-fructan 6-beta-D-fructosyltransferase, FTase) that is capable of synthesizing branched fructan (levan) of high molecular mass from sucrose. In addition, the enzyme catalyzes the fructosyl transfer from sucrose to numerous acceptors other than levan, such as water, short-chain acylalcohols, and various mono- and disaccharides. In the present study, molecular genetic approaches have been utilized to confirm the structure of the catalytic site of Z.mobilis FTase (sucZE2), as follows ; 1) Cloning and characterization of the gene cording for FTase, 2) Presumption of function domains in FTase by random mutation, 3) Alteration of transfructosylating activity by site-directed mutagenesis, 4) Alteration of substrate specificities of FTase by site-directed mutagenesis, 5) Identification of catalycally important residues, Glu278, His296, inFTase gene, 6) Presumption of transfructosylating reaction of FTase via a general acid-base catalytic mechanism.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (42 results)

All Other

All Publications (42 results)

  • [Publications] Hideshi Yanase: "Cloning and characterization of Zymomonas mobilis genes encoding extracellular levansucrase and invertase" Biosci. Biotech. Biochem.59. 197-202 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Cloning Sequence analysis, and expression of the gene encoding formaldehyde dismutase from Pseudomonas putida F61" Biosci. Biotech. Biochem.59. 289-293 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "A recombinant human hemoglobin with asparagine 102 (β) substituted by alanine has a limiting low oxygen affinity" Protein Science. 4. 21-28 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Purification, Crystallization, and Characterization of the Extracellular Invertase from Zymomonas mobilis" J. Ferment. Bioeng., 79 (4), 367-369 (1995). 79. 367-369 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Enzymatic preparation of [1,3-13C] dihydroxyacetone phosphate from [13C] methanol and hydroxypyruvate useing the methanol" Appl. Microbiol. Biotechnol.43. 228-234 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "A signal peptide-independent secretory pathway in gram-negative bacteria, Zymomonas mobilis" Bioscience and Bioindustry. 53. 35-36 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Random mutation of Zymomonas mobilis levansucrase gene" Abstract of paper, Annual Meeting of Japan Society for Biosci. Biotech. Agrochem. 69. 86- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Site-directed mutagenesis of levansucrase gene (sucZE2) from Zymomonas mobilis" Abstract of paper, Annual Meeting of Japan Society for Biosci. Biotech. Agrochem.69. 86- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Gene analysis of transcriptional activator (zliE) from Zymomonas mobilis" Abstract of paper, Annual Meeting of the Society for Ferment. Bioeng. Agrochem. 125- (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Cloning and sequence analysis of the gene encoding 3-hexulose-6 phosphate synthase from the methylotrophic bacterium" FEMS Microbiology Letters. 135. 201-205 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Analysis of structure-function relationship in fructosyltransferase (FTase) from Zymomonas mobilis" Abstract of paper, Annual Meeting of Oyotoushitsu-kagaku kai. 3- (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Expression of Zymomonas zliS gene that enhances secretion of an enzyme via non-GSP in Escherichia coli" Abstract of paper, Annual Meeting of the Society for Ferment, Bioeng. 308- (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kiyoshi Kyono, Hideshi Yanase, Kenzo Tonomura, Haruhiko Kawasaki, and Takuo Sakai: "Cloning and characterization of Zymomonas mobilis genes encoding extracellular levansucrase and invertase" Biosci.Biotech.Biochem.59 (2). 289-293 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Hideyuki Noda, Kiyotaka Aoki, Keiko kita, and Nobuo kato: "Cloning Sequence analysis, and expression of the gene encoding formaldehyde dismutase from Pseudomonas putida F61" Biosci.Biotech.Biochem.59 (2). 197-202 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Lois R.Manning, Kim Vandegriff, Robert M.Winslow, and James M.: "ManningA recombinant human hemoglobin with asparagine 102 (beta) substituted by alanine has a limiting low oxygen affinity, reduced marginally by chloride" Protein Science. 4. 21-28 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Mayumi Iwata, Nobuo Kato, Kenzo Tonomura: "Purification, Crystallization, and Characterization of the Extracellular Invertase from Zymomonas mobilis" J.Ferment.Bioeng.79 (4). 367-369 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Yanase, M.Okuda, K.Kita, Y.Sato, K.Shibata, Y.Sakai, N.Kato: "Enzymatic preparation of [1,3-13C] dihydroxyacetone phosphate from [13C] methanol and hydroxypyruvate useing the methanol-assismilating system of methylotrophic yeasts" Appl.Microbil.Biotechnol.43. 228-234 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "A signal peptide-independent secretory pathway in gram-negative bacteria, Zymomonas mobilis Bioscience and Bioindustry" Bioscience and Bioindustry. 53. 35-36 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Masahiro Maeda, Hiroyuki Yagi, Keiko Kita, Nobuo Kato, Kenzo Tonomura: "Random mutation of Zymomonas mobilis levansucrase gene" Abstract of paper, Annual Meeting of Japan Society for Biosci.Biotech.Agrochem.69. 86 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Hiroyuki Yagi, Hiroyuki Yagi, Keiko Kita, Nobuo Kato, Kenzo Tononura: "Sitedirected mutagenesis of levansucrase gene (sucZE2) from Zymomonas mobilis" Abstract of paper, Annual Meeting of Japan Society for Biosci.Biotech.Agrochem.69. 86 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, Aya Tokunaga, Keiichiro Iwasa, Keiko Kita, Nobuo Kato, Kenzo Tonomura: "Gene analysis of transcriptional activator (zliE) from Zymomonas mobilis" Abstract of paper, Annual Meeting of the Society for Ferment.Bioeng.Agrochem.125 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase, K.Ikeyama, R.Mitsui.S.Ra, K.Kita, Y.Sakai, N.Kato: "Cloning and sequence analysis of the gene encoding 3-hexulose-6 phosphate synthase from the methylotrophic bacterium, Methylomlnas aminofaciens 77a, and its expression in Escherichia coli" FEMS Microbiology Letters. 135. 201-205 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Analysis of structure-function relationship in fructosyltransferase (FTase) from Zymomonas mobilis" Abstract of paper, Annual Meeting of Oyotoushitsu-kagaku kai J.3 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Yanase, Keiichiro Iwasa, Kenji Okamoto, Keiko Kita, Kenzo Tonomura: "Expression of Zymomonas zliS gene that enhances secretion of an enzyme via non-GSP in Escherichia coli" Abstract of paper, Annual Meeting of the Society for Ferment.Bioeng.308 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Hideshi Yanase: "Cloning and characterization of Zymomonas mobilis gene encoding extracellular levansucrase and invertase genes" Biosci.Biotech.Biochem.59. 289-293 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Cloning,Sequence analysis,and expression of the gene encoding formadehyde dismutase from Pseudomonas putida F61" Biosci.Biotech.Biochem.59. 197-202 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "A recombinant human hemoglobin with asparagine 102(b)substituted by alanine has a limiting low oxygen affinity,reduced marginally by chloride" Protein Science. 4. 21-28 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Enzymatic Preparation of [1,3-13C]dihydroxyacetone phosphate from[13C]methanol and hydroxypyruvate using the methanol-assimilating system of methylotrophic yeasts" Appl.Microbiol.Biotechnol.43. 228-234 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 簗瀬 英司: "Zymomonas属細菌にみる新しい酵素分泌機構" バイオサイエンスとバイオインダストリー. 53. 35-36 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Purification,Crystallization,and Characterization of the Extracellular invertase from Zymomonas mobilis" J.Ferment.Bioeng.79. 367-369 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Properties of poly(3-hydroxybutyrate)depolymerase from a marine bacterium,Alcaligenes faecalis AE122" Appl.Environ.Microbiol.61. 1727-1730 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Random mutation of levansucrase gene(sucZE2)from Zymomonas mobilis" Abstract of Papers,Annual Meeting of Japan Society for Bioscience,Biotechnology,and Agrochemistry. 69. 86 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Site-directed mutagenesis of levansucrase gene(sucZE2)from Zymomonas mobilis" Abstract of Papers,Annual Meeting of Japan Society for Bioscience,Biotechnology,and Agrochemistry. 69. 86 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Expression of Zymomonas zliS gene that enhaces secretion of a enzyme via non-GSP in Escherichia coli" Abstract of Papers,Annual Meeting of the Society for Fermentation and Bioengineering. 308 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Cloning and sequence analysis of the gene encoding 3-hexulose-6-phospahte synthase from the methlotrophic bacterium,Methylomonas aminofaciens 77a,and its expression in Escherichia coli" FEMS Microbiology Letters. 135. 201-205 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 簗瀬 英司: "フルクトシルトランスフェラーゼ(FTase)の触媒ドメイン解析と糖転移機能改変" 応用糖質科学. (印刷中).

    • Related Report
      1996 Annual Research Report
  • [Publications] Hideshi Yanase: "Cloning and characterization of Zymomonas mobilis genes encoding extracellular levansucrase and invertase" Biosci.Biotech.Biochem.59. 289-293 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hideshi Yanase: "Cloning,Sequence analysis,and expression of the gene encoding formaldehyde dismutase from Pseudomonas putida F61" Biosci.Biotech.Biochem.59. 197-202 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hideshi Yanase: "A recombinant human hemoglobin with asparagine 102 (β) substituted by alanine has a limiting low oxygen affinity,reduced marginally by choloride" Protein Science. 4. 21-28 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hideshi Yanase: "Enzymatic preparation of [1,3-^<13>C] dihydroxyacetone phosphate from [^<13>C] methanol and hydroxypyruvate useing the methanol-assimilating system of methylotrophic yeasts" Appl.Microbiol.Biotechnol.43. 228-234 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 簗瀬 英司: "Zymomonas属細菌にみる新しい酵素分泌機構" バイオサイエンスとインダストリー. 53. 35-36 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hideshi Yanase: "Purification,Crystallization,and Characterization of the Extracellular Invertase from Zymomonas mobilis" J.Ferment Bioeng.79. 367-369 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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