title: Enzymatic assay of D-serine using D-serine dehydratase from Saccharomyces cerevisiae.
abstract: 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.
authors.affiliation: Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Aichi 464-8601, Japan.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Ito
authors.forename: Kei
authors.initials: K
authors.lastname: Takahashi
authors.forename: Tomoko
authors.initials: T
authors.lastname: Naka
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hemmi
authors.forename: Tohru
authors.initials: T
authors.lastname: Yoshimura
mesh_terms: 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
chemical_list: 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