Molecular Cloning, Functional Expression, and Mutagenesis of cDNA Encoding Rye (Secale cereale) Seed Chitinase-c(Biochemistry & Molecular Biology)

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概要

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• We cloned a complete cDNA encoding rye seed chitinase-c, designated RSC-c, by rapid amplification of cDNA end and PCR procedures. The cDNA of RSC-c consists of 1,018 nucleotides and includes an open reading frame encoding a polypeptide of 266 amino acid residues. A recombinant RSC-c was produced by expression in Escherichia coli Origami(DE3) and purified. rRSC-c had almost the same chitinase activity toward glycolchitin and antifungal activity against Trichoderma sp. as the authentic RSC-c did. RSC-c mutants were subsequently constructed and characterized with respect to their chitinase and antifungal activities. Mutation of Glu67 to Gin completely abolished the chitinase activity and diminished the antifungal activity. Considerable decreases in both activities were observed in the mutations of Trp72 and Ser120 to Ala, and Glu89 to Gln. The roles of these residues in the catalytic event of RSC-c are discussed.

• 社団法人日本農芸化学会の論文
• 2002-02-23

著者

• Yanagita T

  Japan Institute Of Oil & Fats Other Foods Inspection Foundation

• ASO yoichi

  Laboratory of Protein Chemistry & Engineering, Kyushu University

• ISHIGURO Masatsune

  Laboratory of Protein Chemistry and Engineering, Kyushu University Graduate School of Bioresource an

• Yanagita Teruyoshi

  Japan Institute Of Oil & Fats Other Foods Inspection Foundation

• Aso Y

  Institute Of Genetic Resources Graduate School Of Bioresources And Bioenvironmental Science Kyushu U

• Ohnuma Takeshi

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci

• YAMAGAMI Takeshi

  Laboratories of Protein Chemistry and Engineering

• OHNUMA Takayuki

  Laboratories of Protein Chemistry and Engineering

• Ishiguro M

  Laboratory Of Insect Genetic Resources Faculty Of Agriculture Kyushu University

• TAIRA Toki

  Department of Bioscience and Biotechnology, Faculty of Agriculture, Ryukyu University

• YAGI Mikako

  Laboratory of Protein Chemistry and Engineering, Graduate School of Bioresource and Environmental Sc

• TAIRA Toki

  Laboratory of Protein Chemistry and Engineering, Graduate School of Bioresource and Environmental Sc

• Ishihara M

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci

• Ishiguro Masatsune

  Laboratory Of Protein Chemistry & Engineering Department Of Genetic Resources Technology Facultr

• Yamagami T

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci

• Taira T

  Department Of Bioscience And Biotechnology Faculty Of Agriculture Ryukyu University

• Yagi Mikako

  Laboratory Of Protein Chemistry And Engineering Graduate School Of Bioresource And Environmental Sci

• Ishiguro Masatsune

  Laboratory of Insect Genetic Resources, Faculty of Agriculture, Kyushu University

• ASO Yoichi

  Laboratory of Genetic and Protein Engineering, Faculty of Agriculture, Kyushu University

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