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1. 发明申请

US20170268033A1 Method of Analyzing L-Tryptophan in Biological Samples, and Kit Used Therein 审中-公开
公开(公告)号：US20170268033A1
公开(公告)日：2017-09-21
申请号：US15608460
申请日：2017-05-30
申请人： Yasuhisa Asano , Masafumi Kameya , Hiroyasu Onaka
发明人： Yasuhisa Asano , Masafumi Kameya , Hiroyasu Onaka
IPC分类号： C12Q1/26
CPC分类号： C12Q1/26 , C12Y104/03
摘要： Disclosed is a method for quantifying L-tryptophan involving a step for mixing a specimen, L-tryptophan oxidase, and water, a step for allowing the obtained reaction solution to stand a predetermined period of time in the presence of oxygen, and a step for measuring the reaction product resulting from action of enzymes present in the reaction solution after allowing to stand. The L-tryptophan oxidase has a given amino acid sequence and has oxidase activity that generates hydrogen peroxide and ammonia by acting on the L-tryptophan in the presence of oxygen and water. The oxidase activity of the L-tryptophan oxidase on the L-phenylalanine is in the range of 0-3% of the oxidase activity thereof on the L-tryptophan, and the L-tryptophan oxidase does not have oxidase activity on protein-constituting amino acids other than L-tryptophan and L-phenylalanine. Also disclosed are a kit used to quantify the L-tryptophan containing L-tryptophan oxidase, and an enzyme sensor using the L-tryptophan oxidase. This method, kit and enzyme sensor use an L-tryptophan-specific enzyme, so are capable of quantifying L-tryptophan even in the presence of other amino acids.

2. 发明申请

US20140357524A1 Method for Quantifying Subject Substance 有权
标题翻译：量化物质的方法
公开(公告)号：US20140357524A1
公开(公告)日：2014-12-04
申请号：US14453189
申请日：2014-08-06
申请人： Yasuhisa Asano , Masafumi Kameya
发明人： Yasuhisa Asano , Masafumi Kameya
IPC分类号： C12Q1/48 , C12Q1/32 , C12Q1/34
CPC分类号： C12Q1/485 , C12Q1/25 , C12Q1/32 , C12Q1/34 , C12Q1/48 , G01N33/6812 , G01N33/6815
摘要： There is provided a method for quantifying a subject substance, of which typical examples are amino acids. The method of the present invention comprises the following steps: the step of allowing an enzyme that can generate pyrophosphate by using adenosine triphosphate (ATP) as a substrate with converting the subject substance to act on the subject substance to generate pyrophosphate; the step of allowing pyruvate pyrophosphate dikinase (PPDK) to act on the generated pyrophosphate in the presence of adenosine monophosphate (AMP) and phosphoenolpyruvate (PEP) to generate ATP, phosphoric acid, and pyruvate; and the step of quantifying the generated pyruvate, and amount of the subject substance is determined on the basis of the obtained amount of pyruvate. According to the present invention, an amino acid in a biological sample containing a lot of various kinds of contaminants such as inorganic phosphoric acid and urea can be conveniently and quickly quantified without being influenced by the contaminants.
摘要翻译：本发明提供了一种定量目标物质的方法，其中典型的实例是氨基酸。本发明的方法包括以下步骤：通过使用三磷酸腺苷（ATP）作为底物，使可以产生焦磷酸盐的酶转化为目标物质以产生焦磷酸盐; 在磷酸腺苷（AMP）和磷酸烯醇丙酮酸（PEP）存在下，使丙酮酸焦磷酸二激酶（PPDK）作用于产生的焦磷酸盐以产生ATP，磷酸和丙酮酸盐的步骤; 并且基于获得的丙酮酸的量来确定定量产生的丙酮酸的步骤和目标物质的量。根据本发明，可以方便快速地定量含有大量各种污染物如无机磷酸和尿素的生物样品中的氨基酸，而不受污染物的影响。

3. 发明授权

US09708639B2 Method for quantifying amino acids with pyrophosphate 有权
公开(公告)号：US09708639B2
公开(公告)日：2017-07-18
申请号：US14453189
申请日：2014-08-06
申请人： Yasuhisa Asano , Masafumi Kameya
发明人： Yasuhisa Asano , Masafumi Kameya
IPC分类号： G01N33/68 , C12Q1/48 , C12Q1/25 , C12Q1/32 , C12Q1/34
CPC分类号： C12Q1/485 , C12Q1/25 , C12Q1/32 , C12Q1/34 , C12Q1/48 , G01N33/6812 , G01N33/6815
摘要： There is provided a method for quantifying a subject substance, of which typical examples are amino acids. The method of the present invention comprises the following steps: the step of allowing an enzyme that can generate pyrophosphate by using adenosine triphosphate (ATP) as a substrate with converting the subject substance to act on the subject substance to generate pyrophosphate; the step of allowing pyruvate pyrophosphate dikinase (PPDK) to act on the generated pyrophosphate in the presence of adenosine monophosphate (AMP) and phosphoenolpyruvate (PEP) to generate ATP, phosphoric acid, and pyruvate; and the step of quantifying the generated pyruvate, and amount of the subject substance is determined on the basis of the obtained amount of pyruvate. According to the present invention, an amino acid in a biological sample containing a lot of various kinds of contaminants such as inorganic phosphoric acid and urea can be conveniently and quickly quantified without being influenced by the contaminants.

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