US 12,559,780 B2 · Method for efficient catalytic synthesis of PAPS based on constructing ATP regeneration system

Abstract

The present disclosure discloses a method for efficient catalytic synthesis of PAPS based on constructing an ATP regeneration system, and belongs to the technical field of bioengineering. Efficient production of PAPS is realized through microbial recombination expression and artificial construction of PAPS bifunctional synthetase. On the basis, an ATP regeneration system coupling with polyphosphate kinase from Corynebacterium glutamicum and Mycobacterium tuberculosis can be used for recovering two byproducts: pyrophosphoric acid and ADP at the same time, the equivalent conversion of a substrate and a product is realized, the PAPS generated in a catalysis system has high purity, and the sulfonic acid group donation in most sulfonic acid transfer reactions can be realized.

Claims4

Dependent claims

A method for synthesizing 3′-phosphoadenosine-5′-phosphosulfate (PAPS), comprising:

incubating adenosine triphosphate (ATP) with adenosine 5′-phosphosulfate kinase (APS kinase), ATP sulfurylase, and polyphosphate kinase,
wherein the nucleotide sequence of the APS kinase is as set forth in SEQ ID NO:2,
wherein the nucleotide sequence of the ATP sulfurylase is as set forth in SEQ ID NO:3,
wherein the APS kinase and the ATP sulfurylase are linked through a sequence with the amino acid sequence set forth in SEQ ID NO:
1,
wherein the nucleotide sequence of the polyphosphate kinase is as set forth in SEQ ID NO:4, and
wherein a conversion efficiency of the ATP into PAPS is from 83% to 95%.

4 more elements

Dependencies are inferred from the claim text.

Facts

Application

18/192,724Filed 30 Mar 2023 · continuation

Priority

CN 202011069179.830 Sep 2020

Published

US 2023/0272444 A131 Aug 2023

Granted

24 Feb 2026

Term ends

Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12P 19/40
Nucleosides: having a condensed ring system containing a six-membered ring having two nitrogen atoms in the same ring, e.g. purine nucleosides
C12N 9/1229
Transferring phosphorus containing groups, e.g. kinases: phosphotransferases with a phosphate group as acceptor
C12N 9/1241
Transferring phosphorus containing groups, e.g. kinases: nucleotidyltransferases
C12N 9/88
Enzymes; Proenzymes; Compositions thereof ; Processes for preparing, activating, inhibiting, separating or purifying enzymes: lyases

Method for efficient catalytic synthesis of PAPS based on constructing ATP regeneration system

Life of this patent

Method for efficient catalytic synthesis of PAPS based on constructing ATP regeneration system

Life of this patent

Abstract

Claims4