کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
70883 48850 2010 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A highly sensitive enzymatic assay for d- and total serine detection using d-serine dehydratase from Saccharomyces cerevisiae
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی کاتالیزور
پیش نمایش صفحه اول مقاله
A highly sensitive enzymatic assay for d- and total serine detection using d-serine dehydratase from Saccharomyces cerevisiae
چکیده انگلیسی

d-Serine acts as a co-agonist of the N-methyl-d-aspartate (NMDA) receptor, an excitatory glutamate receptor in the mammalian brain that is thought to be involved in higher brain functions such as memory and study. A relationship between the concentration of d-serine in cerebrospinal fluid and neurological disorders such as schizophrenia and amyotrophic lateral sclerosis has been hypothesized. A simple and accurate d-serine assay system would be useful in the study and diagnosis of these diseases. Previously, we developed an enzymatic assay of d-serine using d-serine dehydratase from Saccharomyces cerevisiae, lactate dehydrogenase and NADH. Although this method can detect 10 μM d-serine, a 10-fold increase in sensitivity is required for the assay to be useful for the study and diagnosis of neurological disorders. In this study, we increased the sensitivity of this assay to detect submicromolar concentrations of d-serine. In the new assay, d-serine dehydratase converts d-serine to pyruvate, which is in turn oxidized by pyruvate oxidase. Then, in the presence of horseradish peroxidase, hydrogen peroxide formed during the oxidation converts 10-acetyl-3,7-dihydroxy-phenoxazine (Amplex® Red) to resorufin, which exhibits a strong fluorescence. We show that this improved assay can be used to determine the concentration of d-serine in calf serum.

Figure optionsDownload as PowerPoint slideResearch highlights▶ A sensitive enzymatic assay method for submicromolar d-serine was constructed. ▶ The system consists of the d-serine dehydratase, pyruvate oxidase and peroxidase. ▶ The method can be used for the total serine assay with a mutant alanine racemase. ▶ The method could accurately measure d-serine and total serine in calf serum.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Molecular Catalysis B: Enzymatic - Volume 67, Issues 1–2, November 2010, Pages 150–154
نویسندگان
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