Method for producing 2-keto-3-deoxygluconate from 2-(acetylamino)-2-deoxy-d-gluconic acid by two enzymes

Patent metadata

Field	Value
Publication number	US-11946092-B2
Application number	US-202218051024-A
Country	US
Kind code	B2
Filing date	Oct 31, 2022
Priority date	Apr 8, 2022
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

A method for producing 2-keto-3-deoxygluconate (KDG) from 2-(acetylamino)-2-deoxy-D-gluconic acid (GlcNAc1A) by two enzymes; GlcNAc1A is converted to KDG by incubating GlcNAc1A with a deacetylase OngB at 25° C. for 4-12 h and then with a deaminase OngC at 25° C. for another 10-15 h; it constructs two engineered E. coli/pET22b-ongB (carrying the ongB gene) and E. coli/pET22b-ongC (carrying the ongC gene) strains to prepare recombinant proteins OngB and OngC, respectively; at the action of these two enzymes, OngB and OngC, GlcNAc1A is converted to KDG, which solves the bottleneck of GlcNAc1A utilization during the bioconversion of chitin; the KDG is an important metabolic intermediate to synthesize furan derivatives, herbicides, food additives and other industrially important chemical compounds, having wide industrial applications.

First claim

What is claimed is:

1. A method for producing 2-keto-3-deoxygluconate (KDG) from 2-(acetylamino)-2-deoxy-D-gluconic acid (GlcNAc1A), wherein said method comprises contacting GlcNAc1A with a deacetylase having the amino acid sequence of SEQ ID NO: 3 and a deaminase having the amino acid sequence of SEQ ID NO: 4 for 1-24 h to generate KDG.

2. The method according to claim 1 , wherein the method for producing KDG is carried out by the successive addition of the deacetylase and the deaminase, wherein GlcNAc1A is first incubated with the deacetylase at 25° C. for 12 h, and then with the deaminase at 25° C. for another 12 h.

3. The method according to claim 1 , wherein the molar ratio of the deacetylase to GlcNAc1A is 1:900-1:1100.

4. The method according to claim 1 , wherein the molar ratio of the deaminase to the deacetylase is 1:9-1:11.

5. The method according to claim 1 , wherein the deacetylase is encoded by a deacetylase DNA having the nucleotide sequence of SEQ ID NO: 1.

6. The method according to claim 1 , wherein the deaminase is encoded by a deaminase DNA having the nucleotide sequence of SEQ ID NO: 2.

Assignees

• Univ Shandong

Inventors

• Zhang Yuzhong
• Li Pingyi
• Jiang Wenxin
• Chen Xiulan
• Zhang Yishuo
• Song Xiaoyan

Classifications

• C12P7/42 Primary

  Hydroxy-carboxylic acids · CPC title

• C12N9/78

  acting on carbon to nitrogen bonds other than peptide bonds (3.5) · CPC title

• C12N15/70

  Vectors or expression systems specially adapted for E. coli · CPC title

• C12Y305/01 Primary

  in linear amides (3.5.1) · CPC title

• C12Y305/04

  in cyclic amidines (3.5.4) · CPC title