Enzymatic assay of D-serine using D-serine dehydratase from Saccharomyces cerevisiae.
D-Serine is localized in the mammalian forebrain and modulates brain functions as a coagonist of an N-methyl-D-aspartate receptor. D-Serine is also found in human urine, although its physiological meaning is unclear. A method for rapid and simple assay of D-serine is probably useful for studying its physiological role and clinical relevance. Currently, D-serine is assayed by high-performance liquid chromatography after derivatization of the amino acid to a diastereomer. The method is time consuming and requires expensive equipment. In this study, we developed a rapid and simple method for the D-serine assay using D-serine dehydratase newly found in Saccharomyces cerevisiae. The yeast d-serine dehydratase acts dominantly on d-serine, in contrast with previously reported bacterial enzymes that act on both D- and L-serine. In our method, pyruvate produced from D-serine by the dehydratase reaction is assayed with lactic dehydrogenase and reduced nicotinamide adenine dinucleotide or with 2,4-dinitrophenylhydrazine. Our enzymatic method could be used for the quantitative determination of D-serine in human urine.
Ito
Tomokazu
T
Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi 464-8601, Japan.
Takahashi
Kei
K
Naka
Tomoko
T
Hemmi
Hisashi
H
Yoshimura
Tohru
T
D000411:Alanine Racemase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006836:Hydro-Lyases / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D019289:Pyruvic Acid; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012694:Serine / Q000378:metabolism / Q000652:urine*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013053:Spectrophotometry; D013237:Stereoisomerism
D009243:NAD; D012710:Serum Albumin, Bovine; D012694:Serine; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D006836:Hydro-Lyases; C101950:D-serine dehydratase; D000411:Alanine Racemase