公开(公告)号：US09695460B2

公开(公告)日：2017-07-04

申请号：US14016408

申请日：2013-09-03

申请人： Toyama Prefecture ,  Ajinomoto Co., Inc.

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. 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.

公开(公告)号：US20130344526A1

公开(公告)日：2013-12-26

申请号：US14016408

申请日：2013-09-03

申请人： Ajinomoto Co., Inc. ,  Toyama Prefecture

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.

摘要翻译： 本发明公开了一种定量L-色氨酸的方法，其涉及将样品，L-色氨酸氧化酶和水混合的步骤，使得所得反应溶液在氧气存在下放置预定时间的步骤，以及用于 测量在静置后在反应溶液中存在的酶的作用产生的反应产物。 L-色氨酸氧化酶具有给定的氨基酸序列，具有通过在氧和水存在下作用于L-色氨酸而产生过氧化氢和氨的氧化酶活性。 L-色氨酸氧化酶对L-苯丙氨酸的氧化酶活性在L-色氨酸上的氧化酶活性的0-3％的范围内，并且L-色氨酸氧化酶对构成蛋白质的氨基没有氧化酶活性 L-色氨酸和L-苯丙氨酸以外的酸。 还公开了用于定量含有L-色氨酸的L-色氨酸氧化酶的试剂盒，以及使用L-色氨酸氧化酶的酶传感器。 这种方法，试剂盒和酶传感器使用L-色氨酸特异性酶，因此即使在其他氨基酸存在下也能够定量L-色氨酸。