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Production of novel oligosaccharides by microbes and evaluation of their function

Research Project

Project/Area Number 07556087
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 応用微生物学・応用生物化学
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

TOMITA Fusao  Hokkaido Univ., Fac.of Agr., Prof., 農学部, 教授 (60217536)

Co-Investigator(Kenkyū-buntansha) SAYAMA Koji  NIPPON BEET SUGAR MFG., CO., LTD., Reserch Center, Director, 総合研究所, 所長
HARA Hiroshi  Hokkaido Univ., Fac.of Agr., Associate Prof., 農学部, 助教授 (70198894)
KASAI Takanori  Hokkaido Univ., Fac.of Agr., Prof., 農学部, 教授 (80001444)
YOKOTA Atsushi  Hokkaido Univ., Fac.of Agr., Associate Prof., 農学部, 助教授 (50220554)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 1997: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1996: ¥7,200,000 (Direct Cost: ¥7,200,000)
KeywordsInulin / Levan / Arthrobacter / DFAIII / DFAIV / Inulotriose / Carsium absorption / Mineral absorption / オリゴ糖 / Anthrobacter / DFAIII分解酵素 / DFAIII合成酵素 / 腸内細菌 / Bifidobacterium
Research Abstract

1.Preparation of Raw Material for Oligosaccharides Production : Extraction and purification of inulin from chicory was attempted according to the method of sugar purification from beet, and was successfully achieved. Preparation of levan was also carried out by using levan producing bacteria, and levan was recovered and purified from its culture broth. More simple purification process for preparation of raw material has attempted by such method as addition of oligosaccharide-producing enzyme to crude extracts.
2.Screening and Improvement of Oligosaccharide Producing Microbe : Oligosaccharide producing microbe was screened from plant in addition to soil as a screening source, and xylobiose-producing fungi was newly isolated. Molecular cloning of ift and lft gene, which encode DFAIII and DFAIV producing enzyme respectively, were executed. Expression systems of ift and lft gene was constructed in E.coli system, and higher activities were gained by using these system rather these original strains.
3.Establishment of Oligosaccharide Production Process : Culture conditions for oligosaccharides producing enzymes and reaction conditions were optimized by characterizing microbes and enzymes producing DFAIII,DFAIV,inulotriose and levanbiose. Large scale preparations of purified DFAIII and DFAIV were gained by the process such as fermentation of reaction mixturi by yeast, ion exchange column and crystallization.
4.Evaluation of Physiological Function of Oligosaccharide : Properties of DFAIII with respect to both industrial processing and nutritional function by using microbe, rat and human were examined. In the case of rat, calcium absorption was increased by adding DFAIII to feed, and the regions of absorption were determined in large and small intestines. Physiological fuction of DFAIV has also been evaluation.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] A.Yokota: "Production of inulotriose from invlin by inulin-degrading enzyme from streptomyces rochei E87." Letters in Applied Microbiology. 21. 330-333 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Sakurai: "Molecular cloning of an Inulin tructotransferese(Depolymerisny)Gene from Arthrobacter sp・H65-7 and Its Expression in Escherihiocoli." Biosci.Biotech.Biochem.61(1). 87-92 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Sakurai: "Metabolism of DFAIII by Arthrobacter sp H65-7:Purification and Properties of a DFAIII Hydrolysis Enzyme(DFAIII ase)" Biosci.Biotech.Biochem.61(6). 989-993 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Saito: "Purification of Levan Fructotransferase from Arthrobacter nicotinovorans GS-9 and Production of DFAIV from Levan by the Enzyme." Biosci.Biotech.Biochem.61(10). 1705-1709 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Saito: "Molecular Cloning of Levan Fructoransferase Gene from Arthrobacter nicotinovorans GS-9 and Its Expression in Escherihia coli." Biosci.Biotech.Biochem.61(12). 2076-2079 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 櫻井 博章: "イヌリン-新糖質資源としての展望" バイオサイエンスとインダストリー. 55. 276-278 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Suzuki: "Effects of Difructose Anhydride III on Cakium Absorption in small and Large Intestines of Rats." Biosci.Biotech.Biochem.62(5)(in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] A.Yokota, O.Yamauchi, and F.Tomita: "Production of inulotriose from inulin by inulin-degrading enzyme from Streptomyces rochei E87." Letters in Applied Microbiology. 21. 330-333 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Sakurai, A.Yokota, and F.Tomita: "Molecular Cloning of an Inulin Fructotransferase (Depolymerizing) Gene from Arthrobacter sp. H65-7 and Its Expression in Escherihia coli." Biosci.Biotech.Biochem.61(1). 87-92 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Sakurai, A.Yokota, Y.Sumita, Y.Mori, H.Matsui, and F.Tomita: "Metabolism of DFAIII by Arthrobacter sp. H65-7 : Purification and Properties of a DFAIII Hydrolysis Enzyme (DFAIIIase)." Biosci.Biotech.Biochem.61 (6). 989-993 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Saito, H.Goto, A.Yokota, and F.Tomita: "Purification of Levan Fructotransferase from Arthrobacter nicotinovorans GS-9 and Production of DFAIV from Levan by the Enzyme." Biosci.Biotech.Biochem.61 (10). 1705-1709 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Saito, A.Yokota, and F.Tomita: "Molecular Cloning of Levan FructotransferaseGene from Arthrobacter nicotinovorans GS-9 and Its Expression in Escherihia coli." Biosci.Biotech.Biochem.61 (12). 2076-2079 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Sakurai, A.Yokota, and F.Tomita: "A new prespective on inulin as a carbohydrate resourse." BIOSCIENCE & INDUSTRY. 55. 276-278 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Suzuki, H.Hara, T.Kasai, and F.Tomita: "Effects of Difructose Anhydride III on Calcium Absorption in Small and Large Intestines of Rats." Biosci.Biotech.Biochem.62 (5) (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] A.Yokota: "Production of inulotriose from inulin by inulin degrading enzyme from streptomyces roche:E87" Letters in Applied Microbiology. 21. 330-333 (1995)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Sakurai: "Molecular Cloning of an Inulin Fructotranoferase(Depolymerising)Gene from Arthrobacter sp.H65-7 and Its Expression in Escheribia coli" Biosci.Biotech.Biochem.61. 87-92 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Sakurai: "Metabolism of DFAIII by Arthrobacter sp H65-7:Purification and Properties of a DFAIII Hydrolysis Enzyme(DFAIIIase)" Biosci.Biotech.Biochem.61. 989-993 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Saito: "Purification of Levan Fructotransferase from Arthrobacter nicotinovorans GS-9 and Production of DFAIV from Levan by the Enzyme" Biosci.Biotech.Biochem.61. 1705-1709 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Saito: "Molecular Cloning of Levan Fructotransferase Gene from Arthrobacter nicotinovorans GS-9 and Its Expression in Escherihia coli" Biosci.Biotech.Bichem.61. 2076-2079 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 櫻井博章: "イヌリン-新等質資源としての展望" バイオサイエンスとインダストリー. 55. 276-278 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Suzuki: "Effects of Difructose Anhydride III on Calcium Absorption in Small and Large Intestines of Rats" Biosci.Biotech.Biochem.62(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 櫻井 博章: "イヌリン-新糖質資源としての展望" バイオサイエンスとインダストリー. 55. 25-29 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hiroaki Sakurai: "Molecular Cloning an Inulin Fructotransferase (Depolymerizing) Gene from Arthrobacter sp. H65-7 and Its Expression in Escherichia coli" Biosclence, Biotechnology and Biochemistry. 61. 87-92 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Yokota: "Production of inulotriose from inulin by inulin-degrading enzyme from Streptomyces rochei E87" Letters in Applied Microbiology. 21. 330-333 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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