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Structure and gene of novel glucose isomerase from Bifidobacteria

Research Project

Project/Area Number 03806018
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 応用微生物学・発酵学
Research InstitutionGifu University

Principal Investigator

HORITSU Hiroyuki  Gifu Univ.,Dept.of Agric.,Professor, 農学部, 教授 (60021680)

Co-Investigator(Kenkyū-buntansha) KAWAI Keiichi  Gifu Univ.,Dept.of Agric.,Professor, 農学部, 教授 (00002064)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1992: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsGlucose isomerase / Xylose isomerase / Bifidobacterium adolescentis / Bifidobacterium / Bifidobacterium adolescenti / グルコース・6リン酸イソメラーゼ / ビフイズス菌 / グルコ-ス・イソメラ-ゼ
Research Abstract

To date, glucose isomerase(D-xylose ketol isomerase EC:5.3.1.5)is the enzyme that catalyzes the isomerization of both D-xylose and D-glucose to D-xylulose and D-fructose,respectively. An enzyme that catalyzes only D-glucose into D-fructose is not yet known. For the first time,I succeeded in isolating, purifying and characterizing a novel enzyme from Bifidobacterium adolescentis that is able to catalyze only the isomerization of D-glucose to D-fructose. To produce the enzyme,the bacterium was cultivated anaerobically in the presence of glucose as carbon source at 37゚C for 20 hr. Thereafter,cells(harvested by continuous centrifugation at 12,000 rpm)were disrupted by ultra-sonication and centrifuged at 12,000 rpm for 30 min, The resulting supernatant was then used as crude preparation for purification. The enzyme was purified in 5 step,namely ion exchange chromatography on DE-32 column; Gel filtration chromatography on HW-55-S Toyopearl column; ion exchange chromatography using Smart system with MonoQ-PC16/5 column and HPLC gel filtration chromatography on 300SWxl column. The purified enzyme was characterized and the following are some of its characteristics. (1) Molecular weight was found to be 240,000 by gel filtration method. (2) Optimal pH was 7.0 and the enzyme was stable between pH 6 to 8. (3) Optimal temperature of the enzyme activity was found to be at 40-50゚C,and the thermal stability for the enzyme was found to be up to 40゚C. (4) The effect of metal ion on the enzyme activity indicated that Mg^<2+> and Mn^<2+> were effective. (5) The Km value(D-glucose)of the enzyme was found to be 8mM. This value was about 2 times higher than that exhabited by xylose isomerase. These experimental data prompt me to say that this is a novel enzyme never characterized before.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] 堀津浩章,川合芳文,小西宏明: "ビフィズス菌由来のグルコース・イソメラーゼに関する研究" 岐阜大学地域協同研究センター研究成果報告書. 1. 19-23 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Horitsu,Y.Kawai,K.Konishi & K.Kawai: "D-Glucose isomerase from Bifidobacterium adolescetis" Biosci.Biotoch.Biochem.56. 165-166 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Horitsu.Y.Kawai,H.Konishi & K.Kawai: "D-Glucose isomerase from Bifidobacterium adolescentis" Biosci.Biotech.Biochem.56.(1). 165-166 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 堀津 浩章、川合 芳文、小西 宏明: "ビフイズス菌由来のグルコース・イソメラーゼに関する研究" 岐阜大学地域共同研究センター成果報告書. 1. 19-23 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Horitsu,Y.Kawai,H.Konishi and K.Kawai: "D-Glucose isomerase from Bifidobacterium adolescentis" Biosci.Biotech.Biochem.56. 165-166 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Horitsu,Y.Kawai,H.Honishi&K.Kawai: "DーGlucose isomerase from Bifidobacterium adolescentis" Bioscience,Biotechnology&Biochemistry. 56. 165-166 (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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