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Protein glycosylation and its significance in fungi

Research Project

Project/Area Number 09044214
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research Field 応用微生物学・応用生物化学
Research InstitutionNAGOYA UNIVERSITY

Principal Investigator

TSUKAGOSHI Norihiro  NAGOYA UNIVERSITY,DEPARTMENT OF AGRICULTURE,PROFESSOR, 農学部, 教授 (50115599)

Co-Investigator(Kenkyū-buntansha) HEMMING F.W.  ノッチンガム大学, 生化学部, 教授
PEBERDY J.F.  ノッチンガム大学, 生命科学部, 教授
KATO Masashi  NAGOYA UNIV., DEPT.OF AGRICULTURE,ASSISTANT PROFESSOR, 農学部, 助手 (70242849)
KOBAYASHI Tetsuo  NAGOYA UNIV., DEPT.OF AGRICULTURE,ASSOCIATE PROFESSOR, 農学部, 助教授 (20170334)
JOHN F. Peberdy  NOTTINGHAM UNIV., DEPT.OF LIFE SCIENCE,PROFESSOR
FRANK W. Hemming  NOTTINGHAM UNIV., DEPT.OF BIOCHEMISTRY,PROFESSOR
PEBARDY J.F.  ノッチンガム大学, 生命科学部, 教授
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
KeywordsA.oryzae / A.nidulans / Glycan structure of Taka-amylase A, (TAA) / Glycan structure of AoTAA / Glycan structure of AnTAA / Difference in glycosylation between A.oryzae and A.nidulans / タカミラーゼA(TAA)糖鎖
Research Abstract

In order to elucidate the significance of protein glycosylation during protein secretion and in expression of enzymatic function, the glycosylation mechanism in filamentous fungi has been investigated molecular-biologically and biochemically using Taka-amylase A as a model enzyme. Our joint works carried out at both sites during last two years are summarized as follows ;
1) The A.oiyzae Taka-amylase A (AoTAA) and Taka-amylase A produced by A.nidulans carrying the A.ozyzae Taka-amylase A gene (AnTAA) were highly purified to homogeneity from commercially available enzyme preparation and the culture broth of an A.nidulans trasformant, respectively.
2) The glycan structures of AoTAA and AnTAA were analyzed by lectin blotting. Both enzymes contained mannose as a component of gylcans. However, a highly glycosylated protein with a low molecular mass was found to be contaminated in those preparations during this analysis. This low-molecular-mass glycoprotein interfered further monosaccharide analysis of the glycans. Although we made our best to get rid of this contaminant from AnTAA preparations using several purification methods, we have not yet succeed to remove the contaminant from the enzymes.
3) The glycan structures were analyzed by fluorescent labeling technique and we found that AoTAA and AnTAA comprised 7 and S monosaccharides, respectively. This indicates the glycosylation process is different in A.oryzae and A.nidulans

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Masashi Kato: "An Aspergillus nidulans nuclear protein, AnCP, involved in enhancement of Taka-amylase A gene expression,binds to the CCAAT-containing taaG2, amdS and gatA promoters." Mol.Gen.Genet.254. 119-126 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kato: "The Aspergillus nidulans CCAAT-binding factor, AnCP/AnCF, is a heteromeric protein analogous to the HAP complex of Saccharomyces cerevisiae." Mol.Gen.Genet.257. 404-411 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Norihuki Kitamoto: "Utilization of the TEF1-α gene (TEF1) promoter for expression of polygalacturonase genes, pgaA and pgaB, in Aspergillus oryzae." Appl.Microbiol.Biotechnol.50. 85-92 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kayoko Takeda: "Cloning of a thermostable ascorbate oxidase from Acremonium sp.HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis." Biochimica Biophysica Acta. 1388. 444-456 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kayoko Takeda et al: "Cloning of a thermostable ascorbate oxidase gene from Acremonium sp.HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis." Bichim.Biophys.Acta. 1388. 444-456 (1398)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kato et al.: "An Aspergillus nidulans nuclear protein, AnCP,involved in enhancement of Taka-amylase A gene expression, binds to the CCAAT-containg taaG2, amdS and gatA promoters" Mol.Gen.Genet.254. 119-126 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kato et al.: "The Aspergillus nidulans CCAAT-binding factor, AnCP/AnCF,is a heteromeric protein analogous to the HAP complex of Saccharmyces cerevisiae." Mol.gen.Genet.257. 404-411 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Noriyuki Kitamoto et al.: "Utilization of the TEF1-alpha gene (TEF1 )promoter for expression of polygalacturonase genes, pgaA and pgaB,in Aspergillus oryzae." Appl.Microbiol.Biotechnol.50. 85-92 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kato: "An Aspergillus nidulans nuclear protein,AnCP,involved in enMancement of Taka-amylase A gene expression,binds to the CCAAT-containing taaG2,amdS and gatA promoters." Mol.Gen.Genet.254. 119-126 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masashi Kato: "The Aspergillus nidulans CCAAT-binding factor,AnCP/AnCF,is a heteromeric protein analogous to the HAP complex of Saccharomyces cerevisiae." Mol.Gen.Genet.257. 404-411 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Norihuki Kitamoto: "Utilization of the TEF1-α gene(TEF1) promoter for expression of polygalacturonase genes,pgaA and pgaB,in Aspergillus oryzae." Appl.Microbiol.Biotechnol.50. 85-92 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kayoko Takeda: "Cloning of a thermostable ascorbate oxidase from Acremonium sp.HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis." Biochimica Biophysica Acta. 1388. 444-456 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masashi Kato: "The Aspergillus nidu lans CCAAT-binding factor,AnCP/AnCF,is a heteromeric protein analogous to the HAP complex of Soccharomyces cerevisiae." Mol.Gen.Genet.(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masashi Kato: "An Aspergillus nidulans nuclear protein.AnCP,involved in enhancement of Takaamylase Asone expression binds to the CCAAT-contoining taaG2,and S and gatA promoters." Mol.Gen.Genet.254. 119-126 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masashi Kato: "Seguence,specific binding sites in the Taka-amylase AG2 promotor for the CreA repressor mediating carbon catabolite repression." Biosci.Biotech.Biochem.60. 1776-1779 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Noriyuki Kitamoto: "Molecular cloning,purification and characterization of two endo-1,4-gluconases from Aspergillus oryzae KBN616." Appl.Microbiol.Biotechnol.46. 538-544 (1996)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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