US 8,785,147 B2 · L-threonine analysis method and L-threonine dehydrogenase

Abstract

A method for analyzing L-threonine contained in an specimen, which includes the steps of mixing a sample containing the specimen with an L-threonine dehydrogenase derived from Cupriavidus necator and a coenzyme NAD+ and analyzing the amount of NADH or 2-amino-3-oxobutyric acid after a predetermined period; an L-threonine dehydrogenase derived from Cupriavidus necator, which is a novel L-threonine dehydrogenase (TDH; EC 1.1.1.103) and can be utilized in the above-mentioned analysis method; a method for preparing a gene or the like to be used in the preparation of the enzyme, or a method for preparing the enzyme; an L-threonine analysis kit which includes (A) the L-threonine dehydrogenase and (B) a coenzyme NAD+; an enzyme preparation for use in the analysis of L-threonine, which includes the L-threonine dehydrogenase contained in a buffer solution; and an enzyme sensor utilizing the L-threonine dehydrogenase.

Claims5

Dependent claims

A method for analyzing the L-threonine contained in a specimen, comprising:

A) mixing a sample comprising a specimen and L-threonine dehydrogenase, with the coenzyme NAD+;
and
B) determining the quantity of a product after a time period,
wherein the product is selected from the group consisting of NADH and 2-amino-3-oxybutyric acid;
wherein said L-threonine dehydrogenase has L-threonine dehydrogenase activity that is oxidative and specific for L-threonine to 2-amino-3-oxobutyric acid, and is selected from the group consisting of:
(i) L-threonine dehydrogenase from Cupriavidus necator;
(ii) a protein comprising the amino acid sequence of SEQ ID NO:
1;
(iii) a protein comprising the amino acid sequence of SEQ ID NO:
1, but having from 1 to 30 amino acid deletions, substitutions, and/or additions;
and
(iv) a protein comprising an amino acid sequence having 90% or greater homology with the amino acid sequence of SEQ ID NO:
1.

10 more elements

Dependencies are inferred from the claim text.

L-threonine analysis method and L-threonine dehydrogenase

Life of this patent

L-threonine analysis method and L-threonine dehydrogenase

Life of this patent

Abstract

Claims5