Heterologous Expression of Dehydroascorbate Reductase from Rice and Its Application to Determination of Dehydroascorbate Concentrations

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概要

• 論文の詳細を見る

• We constructed an expression vector for rice dehydroascorbate reductase (DHAR) (EC 1.8.5.4) with a polyhistidine tag at the amino terminus and introduced the vector into several strains of Escherichia coli. On conventional induction treatment with isopropylthiol-β-D-galactoside, E. coli harboring rice DHAR cDNA produced doublet polypeptides of about 27 kDa. Induction duration or growth temperature did not affect the ratio of these polypeptides. Only the larger polypeptide, corresponding to full-length recombinant DHAR, was produced in E. coli supplemented with tRNAs for several minor codons. Most of the enzymatic characteristics of the recombinant DHAR were similar to those of the native one, although the recombinant protein showed increased heat susceptibility. Using recombinant DHAR, we developed a method for simple and precise determination of dehydroascorbate concentrations in tissue extracts by spectrophotometry, and we successfully applied the method to several fruit juices and vegetables.

• 日本ビタミン学会の論文

著者

• USHIMARU Takashi

  Department of Biology and Geoscience, Faculty of Science, Shizuoka University

• Ushimaru Takashi

  Department Of Biology And Geosciences Faculty Of Science Shizuoka University

• Ushimaru Takashi

  Department Of Biology And Geoscience Faculty Of Science Shizuoka University

• Goda Kiyoshi

  Faculty Of Nutrition Kobe Gakuin University:high Technology Research Center Kobe Gakuin University

• Ogishi Yasuka

  Faculty Of Nutrition Kobe Gakuin University

• Amako Katsumi

  Faculty Of Nutrition Kobe Gakuin University:high Technology Research Center Kobe Gakuin University

• Ishikawa Ayami

  Faculty Of Nutrition Kobe Gakuin University

• KISHIMOTO Ritsuko

  Faculty of Nutrition, Kobe Gakuin University

• Kishimoto Ritsuko

  Faculty Of Nutrition Kobe Gakuin University

• Goda Kiyoshi

  Faculty Of Nutrition Kobe Gakuin University

• Ishikawa Ayumi

  Faculty of Nutrition, Kobe Gakuin University

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