Spectrophotometric assay of d-isoleucine using an artificially created d-amino acid dehydrogenase

Hironaga Akita, Yoshifumi Imaizumi, Hirokazu Suzuki, Katsumi Doi, Toshihisa Ohshima

研究成果: ジャーナルへの寄稿記事

5 引用 (Scopus)

抄録

d-isoleucine (d-Ile) can be assayed using chiral chromatography but the availability of that method is limited by the necessity for special expertise and expensive equipment. We therefore developed a simple and specific colorimetric assay system for d-Ile determination using an artificially created NADP+-dependent d-amino acid dehydrogenase (DAADH). The system consists of two reaction steps: the first is the quantitative conversion of d-Ile to (3R)-2-oxo-3-methyl valerate by DAADH in which NADP+ is converted to NADPH, while the second is chemical conversion of NADPH to reduced water-soluble Tetrazolium-3 via a redox mediator. d-Ile was determined from 1 to 50 µM, and the assay was unaffected by the presence of any of three other isomers (100 µM), alcohol and organic acids.

元の言語英語
ページ(範囲)2245-2248
ページ数4
ジャーナルBiotechnology letters
36
発行部数11
DOI
出版物ステータス出版済み - 1 1 2014

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Isoleucine
NADP
Amino acids
Assays
Oxidoreductases
Amino Acids
Organic acids
Chromatography
Isomers
Alcohols
Availability
Valerates
Oxidation-Reduction
Water
Equipment and Supplies
Acids

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

これを引用

Spectrophotometric assay of d-isoleucine using an artificially created d-amino acid dehydrogenase. / Akita, Hironaga; Imaizumi, Yoshifumi; Suzuki, Hirokazu; Doi, Katsumi; Ohshima, Toshihisa.

:: Biotechnology letters, 巻 36, 番号 11, 01.01.2014, p. 2245-2248.

研究成果: ジャーナルへの寄稿記事

Akita, Hironaga ; Imaizumi, Yoshifumi ; Suzuki, Hirokazu ; Doi, Katsumi ; Ohshima, Toshihisa. / Spectrophotometric assay of d-isoleucine using an artificially created d-amino acid dehydrogenase. :: Biotechnology letters. 2014 ; 巻 36, 番号 11. pp. 2245-2248.
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AU - Ohshima, Toshihisa

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