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Molecular Biology and Engineering of Biosynthesis of Sulfur-containing Metabolites in Plants

Research Project

Project/Area Number 08458170
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bioorganic chemistry
Research InstitutionCHIBA UNIVERSITY

Principal Investigator

SAITO Kazuki  Chiba University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (00146705)

Co-Investigator(Kenkyū-buntansha) NOJI Masaaki  Chiba University, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (80271534)
YAMAZAKI Mami  Chiba University, Faculty of Pharmaceutical Sciences, Lecturer, 薬学部, 講師 (70222370)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1998: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsGenetic engineering / Natural products / Moleculart cloning / Biosynthesis / Amino acids / Sulfur metabolism / Cysteine / Alliin
Research Abstract

Cysteine is the first organic precursor for the formation of sulfur-containing metabolites in plants such as methionine and glutathione. Two regulatory mechanisms for cysteine formation have been clarified by our recent studies ; one at the level of enzyme activity and the other at the steady-state mRNA level. The enzyme activity of serine acetyltransferase catalyzing the formation of O -acetylserine from serine and acetyl - CoA is inhibited by L-cysteine at the concentration of less 10 _ M.This inhibition was observed in the isoforms from watermelon, spinach and Arabidopsis thaliana. However, two isozymes from A.thaliana presumably localizing in organelles were insensitive to L-cysteine, suggesting different regulation among isozymes of different subcellular localization. Northern hybridization analysis of sulfate starved plants indicated that the steady-state mRNA level of an isoform of sulfate transporter (AST68) increased specifically in roots up to -9 folds by sulfate starvation Among all the structural genes encoding the proteins for sulfate assimilation, sulfate transporter (AST68) and APS reductase are inducible by sulfate starvation in A.thaliana. The experiments of in situ hybridization and transgenic plants with the promoter-gus fusion indicated that AST68 is expressed in the central cylinder of roots and the vascular tissues of shoots. The transgenic plants of AST68 anti-sense construct exhibited sulfur deficient symptom in cotyledons and also tolerance to selenate. These results indicating that AST68 plays a central role in sulfate assimilation in A.thaliana. The cDNA encoding alliinase responsible for conversion of alliin to sulfenic acid was isolated from A Ilium tuberosum. The recombinant protein expressed in yeast cells exhibited the alliinase activity.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] H.Takahashi: "Regulation of sulfur assimilation in higher plants : A sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana." Proc.Natl.Acad.Sci.USA. 94. 11102-11107 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Manabe: "Alliinase (S-alk(en)yl-L-cysteine sulfoxide lyase) from Allium tuberosum (Chinese chive) : Purification, localization, cDNA cloning and heterologous functional" Eur.J.Biochem.257. 21-30 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M Noji: "Isoform-dependent differences in feedback regulation and subcellular localization of serine acetyltransferase involved in cysteine biosynthesis from Arabidopsis thaliana." J.Biol.Chem.273. 32739-32745 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] C-L.Ho: "Molecular characterization of plastidic phosphoserine aminotransferase in serine biosynthesis from Arabidopsis." Plant J.16. 443-452 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 斉藤和季: "植物の硫黄同化系の分子機構と制御" 化学と生物. 37. 156-163 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hideaki Takahashi, Mami Yamazaki, Noriko Sasakura, Akiko Watanabe, Thomas Leustek, Janice de Almeida Engler, Gilbert Engler, Marc Van Montagu and Kazuki Saito: "Regulation of sulfur assimilation in higher plants : A sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana" Proc.Natl.Acad.Sci.USA. 94. 11102-11107 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tomofumi Manabe, Asako Hasumi, Mitsuyo Sugiyama, Mami Yamazaki and Kazuki Saito: "Alliinase(S-alk(en)yl-L-cysteine sulfoxide lyase)from Allium tuberosum(Chinese chive) : Purification, localization, cDNA cloning and heterologous functional expression." Eur.J.Biochem.257. 21-30 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masaaki Noji, Kenji Inoue, Nobuhito Kimura, Atsuko Gouda and Kazuki Saito: "Isoform-dependent differences in feedbac regulation and subcellular localization of serine acetyltransferase involved in cysteine biosynthesis from Arabidopsis thaliana." J.Biol.Chem.273. 32739-32745 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chai-Ling Ho, Masaaki Noji, Maiko Saito, Mami Yamazaki and Kazuki Saito: "Molecular characterization of plastidic phosphoserine aminotransferase in serine biosynthesis from Arabidopsis." Plant J.16. 443-452 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tomotumi MANABE et al.: "Alliinase[S-alk(en)yl-L-cysteine sulfoxide lyase]from Allium tuberosum (Chinese chive)" Eur.J.Biochem.257. 21-30 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kazuki Saito, et al.,: "Production of plant non-protein amino acids by recombinant enzymes of sequential biosynthetic reactions in bacteria" Biol.Pharm.Bull.20. 47-53 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hideki Takahashi and kazuki Saito: "Subcellular Localization of Spinach Cysteine Synthase Isotorms and Regulation of Their Gene Expression by Nitrogen and Sultur" Plant Physiol.112. 273-280 (1996)

    • Related Report
      1996 Annual Research Report

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

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