US 12,012,619 B2 · Methods of use for an NADPH-regeneration system based on a monomeric isocitrate dehydrogenase

Abstract

An NADPH-regeneration system based on monomeric isocitrate dehydrogenase (IDH) and a use thereof. Specifically, the present invention relates to a recombinant vector including a polynucleotide encoding an isocitrate dehydrogenase recombinant protein derived from Corynebacterium glutamicum (CgIDH) and an isocitrate dehydrogenase recombinant protein derived from Azotobacter vinelandii (AvIDH), a method for producing the recombinant protein, and an NADPH-regeneration system using the recombinant protein produced by the method. The enzyme in a monomeric form that may be efficiently used in the NADPH-regeneration system in the transformant into which the recombinant vector was introduced, was found, and the NADPH-regeneration system using the enzyme in a monomeric form has a very high utility value as biological parts and biocatalyst materials that provides NADPH to the NADPH-dependent enzyme.

Claims6

Dependent claims

A method of using a recombinant expression vector to regenerate NADPH comprising:

producing the recombinant expression vector comprising a polynucleotide encoding a monomeric isocitrate dehydrogenase (IDH) from Corynebacterium glutamicum or Azotobacter vinelandii;
producing a transformant transformed with the recombinant expression vector;
culturing the transformant to express the monomeric IDH;
recovering the soluble expressed monomeric IDH;
and
coupling the recovered monomeric IDH to an NADPH-dependent enzyme,
wherein the transformant is Escherichia coli, wherein the monomeric IDH from Corynebacterium glutamicum consists of the amino acid sequence of SEQ ID NO:
1, and the monomeric IDH from Azotobacter vinelandii consists of the amino acid sequence of SEQ ID NO:
2.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application

18/054,791Filed 11 Nov 2022 · division

Priority

KR 10-2021-002323222 Feb 2021

Published

US 2023/0143972 A111 May 2023

Granted

18 Jun 2024

Term ends

Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12N 9/0006
Oxidoreductases: acting on CH-OH groups as donors
C12N 9/0042
With a heme protein as acceptor: nADPH-cytochrome P450 reductase
C12P 7/02
Preparation of oxygen-containing organic compounds: containing a hydroxy group
C12P 17/165
Containing two or more hetero rings: heterorings having nitrogen atoms as the only ring heteroatoms

Methods of use for an NADPH-regeneration system based on a monomeric isocitrate dehydrogenase

Life of this patent

Methods of use for an NADPH-regeneration system based on a monomeric isocitrate dehydrogenase

Life of this patent

Abstract

Claims6