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    • 92. 发明授权
    • Assay method for amylase activity and method of producing maltose
dehydrogenase for use therein
    • 淀粉酶活性的测定方法和生产用于其中的麦芽糖脱氢酶的方法
    • US4427771A
    • 1984-01-24
    • US311263
    • 1981-10-14
    • Hideo MisakiEiji MuramatsuHidehiko IshikawaKazuo Matsuura
    • Hideo MisakiEiji MuramatsuHidehiko IshikawaKazuo Matsuura
    • C12N9/04C12Q1/40C12N1/20C12Q1/32C12R1/11
    • C12N9/0006C12Q1/40G01N2400/18Y10S435/837
    • An assay method for amylase activity in a biological specimen such as serum, saliva or urine. The enzyme amylase in the specimen is used to decompose a substrate which is a glucose polymer having a modified reducing terminal glucose residue or a cyclic glucose polymer. A component of the decomposed substrate is measured as an indication of amylase activity in the specimen. The residue may be amylose, amylopectin, starch, starch hydrolyzate, an etherified reducing terminal, an esterified reducing terminal, gluconolactone or a gluconic acid residue or its derivative. Decomposed substrate assay may be effected by contacting the same with maltose dehydrogenase and NAD or NADP, whereupon the assay is performed by measuring the amount of reduced NAD or reduced NADP, by reacting the same with reduced-form hydrogen transport colorimetric reaction reagent. This reagent may be a tetrazolium salt and diaphorase, or tetrazolium salt and phenazinemethosulfate. To remove pre-existing glucose and maltose present in the specimen, the specimen may be pretreated with alpha-glucosidase or kinase in the presence of Mg.sup.++ and ATP, the kinase being for example hexokinase. The preferred maltose dehydrogenase is produced by culturing Bacillus megaterium B-0779 FERM-P No. 5662.
    • 生物样品如血清,唾液或尿液中淀粉酶活性的测定方法。 试样中的酶淀粉酶用于分解具有修饰的还原性末端葡萄糖残基或环状葡萄糖聚合物的葡萄糖聚合物的底物。 测量分解的底物的组分作为样品中淀粉酶活性的指示。 残基可以是直链淀粉,支链淀粉,淀粉,淀粉水解物,醚化还原末端,酯化还原末端,葡萄糖酸内酯或葡萄糖酸残基或其衍生物。 分解的底物测定可以通过使其与麦芽糖脱氢酶和NAD或NADP接触来实现,因此通过使其与还原形式的氢转运比色反应试剂反应来测量还原的NAD或还原的NADP的量来进行测定。 该试剂可以是四唑盐和心律黄素,或四唑鎓盐和吩嗪甲硫酸盐。 为了除去样品中存在的现有葡萄糖和麦芽糖,可以在Mg ++和ATP存在下用α-葡糖苷酶或激酶预处理样品,所述激酶例如己糖激酶。 优选的麦芽糖脱氢酶是通过培养巨大芽孢杆菌B-0779 FERM-P No.5662产生的。