US 2019/0094235 A1Published application

Method for quantifying amino acid and amino acid quantification kit

AssigneesIkeda Food Research Co., Ltd., Hiroshima City University

InventorsDaisuke SATO, Tomoko NAKATSUKA, Hideyuki AOKI, Mikiko KIDA, Kenta YAMADA, Shoji KANEKO, Akimitsu KUGIMIYA

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Abstract

[Problem to be solved] It is to provide a method for selectively and easily quantifying the L-form and/or D-form amino acids to be measured using an AARS with high sensitivity, and an amino acid quantification kit. [Solution to Problem] A method for quantifying amino acids (L-AA and/or D-AA) in a sample using an AARS, wherein the amino acids and the AARS are released from an aminoacyl AMP-AARS complex once formed, and they are used again for forming the aminoacyl AMP-AARS complex, so that reaction products such as pyrophosphoric acid to be measured can be ultimately produced up to a molar number larger than that of the amino acids contained in the sample, and an amino acid quantification kit for performing the method.

Claims12

Dependent claims

A method for quantifying amino acids in a sample, comprising:

Step (I) comprising following steps:
(Step I-1) a step including a reaction (Reaction 1) wherein L-form and/or D-form amino acids (L-AA and/or D-AA) in the sample, an aminoacyl tRNA synthetase (AARS) corresponding to the amino acids and an adenosine triphosphate (ATP) are reacted in the presence of a divalent ion or a polyamine to form a complex comprising an aminoacyl adenylate (aminoacyl AMP) and the AARS (aminoacyl AMP-AARS complex);
(Step I-2) a step including a reaction (Reaction 2) wherein an amino acid-regenerating agent acts on the aminoacyl AMP-AARS complex formed in Reaction 1 or Reaction 3 to release the AARS and the amino acids (L-AA and/or D-AA) from the complex;
(Step I-3) a step including a reaction (Reaction 3) wherein the amino acids (L-AA and/or D-AA) released in Reaction 2 and/or the AARS are reused in Reaction 1 to cause the aminoacyl AMP-AARS complex reaction;
and
(Step I-4) a step of repeating the Step I-2 and the Step 1-3, and
Step (II) comprising measuring an amount of reaction products produced in the Step (I) and determining an amount of the L-form and/or D-form amino acids on the basis of the measured amount of the reaction products.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application: 16/145,509Filed 28 Sep 2018
Priority: JP 2017-18738528 Sep 2017
Published: 28 Mar 2019
Term ends: Not estimatedThe source records don’t support an estimate.

Classification4 codes

G01N 33/6812
Determination of free amino acids: assays for specific amino acids
G01N 27/4168
Measuring a particular property of an electrolyte: oxidation-reduction potential, e.g. for chlorination of water
G01N 33/6806
General methods of protein analysis not limited to specific proteins or families of proteins: determination of free amino acids
G01N
Investigating or analysing materials by determining their chemical or physical properties

Family

Application16/145,509Continuation in part
US 10,942,187 B2Related patent
US 2019/0094235 A1This patent

From the US filing history. Foreign family members aren’t listed.

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