METHOD OF MEASURING ADENINE NUCLEOTIDE

• US 20090162881A1
• Filed: 11/10/2006
• Published: 06/25/2009
• Est. Priority Date: 11/14/2005
• Status: Abandoned Application

• Current Assignee: Panasonic Corporation (Panasonic Holdings Corporation), Tokyo University of Agriculture and Technology
• Original Assignee: Matsushita Electric Industrial Company Limited (Panasonic Holdings Corporation), Tokyo University of Agriculture and Technology
• Inventors: Okamura, Daisuke, Tsugawa, Wakako, Sode, Koji
• Application Number:
  US12/093,521
• Publication Number:
  US 20090162881A1
• Time in Patent Office: Days
• US Class Current: 435/15
• CPC Class Codes: C12Q 1/008 for determining co-enzymes or co-factors, e.g. NAD, ATP
• CPC Class Codes: G01N 33/5735 co-enzymes or co-factors, e.g. NAD, ATP

Abstract

A high sensitivity electrochemistry type method for measuring adenine nucleotide which has a convenient and further miniaturized measuring device structure; is low in consumptive power; and does not require a treatment operation for substances that cause turbidity is provided. A method for measuring adenine nucleotide, which comprises a step A for converting adenosine triphosphate to adenosine diphosphate by an enzyme E₁, a step B for converting said adenosine diphosphate and a phosphate donor P₂ to adenosine triphosphate and dephosphorylated phosphate donor P_2′ by an enzyme E₂, and a step C for electrochemically measuring said donor P_2′ by carrying out an oxidation-reduction reaction.

Claims

1. A method for measuring adenine nucleotide, which comprises

a step A for converting adenosine triphosphate to adenosine diphosphate by an enzyme E₁;

a step B for converting said adenosine diphosphate and a phosphate donor P₂ to adenosine triphosphate and dephosphorylated phosphate donor P_2′

by an enzyme E₂; and

a step C for electrochemically measuring said donor P_2′

by carrying out an oxidation-reduction reaction of said donor P_2′

.

2. The method for measuring adenine nucleotide according to claim 1, wherein the adenosine triphosphate and the dephosphorylated phosphate donor P_2′

are formed according to the frequency of the reactions by repeating a cycle consisting of the step A and step B two or more times to carry out a reaction.

3. The method for measuring adenine nucleotide according to claim 1, wherein the dephosphorylated phosphate donor P_2′

formed by step B is measured as the amount of adenine nucleotide.

4. The method for measuring adenine nucleotide according to claim 2, wherein myokinase is used as the enzyme E₁.

5. The method for measuring adenine nucleotide according to claim 1, wherein the oxidation-reduction reaction in the step C is carried out by an oxidation-reduction enzyme E₃.

6. The method for measuring adenine nucleotide according to claim 5, wherein the donor P_2′

is electrochemically measured in the step C by carrying out an oxidation-reduction reaction of said donor P_2′

which consumes oxygen molecule and detecting amount of the consumed oxygen.

7. The method for measuring adenine nucleotide according to claim 5, wherein the donor P_2′

is electrochemically measured in the step C by carrying out an oxidation-reduction reaction of said donor P_2′

which produces hydrogen peroxide and detecting amount of the produced hydrogen peroxide.

8. The method for measuring adenine nucleotide according to claim 5, wherein the electrochemically detecting way in the step C uses an electron mediator as the electron acceptor.

9. The method for measuring adenine nucleotide according to claim 1, wherein pyruvate kinase is used as the enzyme E₂, and pyruvate oxidase is used as the enzyme E₃.

10. The method for measuring adenine nucleotide according to claim 1, wherein pyruvate kinase is used as the enzyme E₂, and pyruvate dehydrogenase is used as the enzyme E₃.

11. The method for measuring adenine nucleotide according to claim 1, wherein hexokinase or glucokinase is used as the enzyme E₂, and glucose oxidase is used as the enzyme E₃.

12. The method for measuring adenine nucleotide according to claim 2, wherein the dephosphorylated phosphate donor P_2′

formed by step B is measured as the amount of adenine nucleotide.

13. The method for measuring adenine nucleotide according to claim 3, wherein myokinase is used as the enzyme E₁.