Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 2245-2248 |
| Number of pages | 4 |
| Journal | Biotechnology letters |
| Volume | 36 |
| Issue number | 11 |
| DOIs | |
| Publication status | Published - Jan 1 2014 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
Cite this
Spectrophotometric assay of d-isoleucine using an artificially created d-amino acid dehydrogenase. / Akita, Hironaga; Imaizumi, Yoshifumi; Suzuki, Hirokazu; Doi, Katsumi; Ohshima, Toshihisa.
In: Biotechnology letters, Vol. 36, No. 11, 01.01.2014, p. 2245-2248.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Spectrophotometric assay of d-isoleucine using an artificially created d-amino acid dehydrogenase
AU - Akita, Hironaga
AU - Imaizumi, Yoshifumi
AU - Suzuki, Hirokazu
AU - Doi, Katsumi
AU - Ohshima, Toshihisa
PY - 2014/1/1
Y1 - 2014/1/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=84920707979&partnerID=8YFLogxK
U2 - 10.1007/s10529-014-1597-z
DO - 10.1007/s10529-014-1597-z
M3 - Article
C2 - 24966047
AN - SCOPUS:84920707979
VL - 36
SP - 2245
EP - 2248
JO - Biotechnology Letters
JF - Biotechnology Letters
SN - 0141-5492
IS - 11
ER -