Alditol oxidase and application thereof

Patent metadata

Field	Value
Publication number	US-2025115937-A1
Application number	US-202318859487-A
Country	US
Kind code	A1
Filing date	Nov 30, 2023
Priority date	Oct 9, 2023
Publication date	Apr 10, 2025
Grant date	—

Abstract

An alditol oxidase and application thereof. The method uses D-glucose as a substrate to dock the alditol oxidase derived from Streptomyces coelicolor A3, and selects amino acid residues around an active center for saturation mutagenesis, and screens for alditol oxidase with D-glucose oxidizing activity by plate color development. An amino acid sequence of the alditol oxidase is shown in SEQ ID NO: 2 or SEQ ID NO: 4. The alditol oxidase has the activity of converting D-glucose to D-gluconic acid and D-glyceraldehyde to D-glyceric acid. By using the alditol oxidase, the conversion of D-glucose to pyruvic acid is realized by only three enzymes for the first time and does not depend on any coenzyme.

First claim

1 . An alditol oxidase, with an amino acid sequence as shown in SEQ ID NO: 2 or SEQ ID NO: 4.

2 . A gene encoding the alditol oxidase of claim 1 .

3 . The gene of claim 2 , wherein the gene has a nucleotide sequence as shown in SEQ ID NO: 3 or SEQ ID NO: 5.

4 . A recombinant vector or a recombinant strain comprising the gene of claim 2 .

5 . The recombinant vector or the recombinant strain of claim 4 , wherein the recombinant vector uses pET28a as an expression vector.

6 . The recombinant vector or the recombinant strain of claim 4 , wherein the recombinant strain uses Escherichia coli BL21 (DE3) as a host.

7 . A method of synthesizing pyruvic acid, comprising using D-glucose as a substrate, and synthesizing pyruvic acid through a catalytic reaction of an alditol oxidase, dehydratase and 2-keto-3-deoxy gluconate aldolase, wherein the alditol oxidase is the alditol oxidase of claim 1 .

8 . The method of claim 7 , wherein final concentration of the alditol oxidase is 0.3˜0.7 mg/mL, final concentration of the dehydratase is 0.3˜0.5 mg/mL, and final concentration of the 2-keto-3-deoxy gluconate aldolase is 0.1˜0.3 mg/mL in a reaction system.

9 . The method of claim 8 , the reaction system further comprising MgCl 2 , Tween 20 and manganese dioxide, wherein reaction temperature of the reaction system is 38˜42° C.

10 . An application of the alditol oxidase of claim 1 in catalyzing oxidation of aldose, wherein the aldose is one or more selected from D-xylose, D-ribose, L-arabinose, D-erythrose, D-mannose and D-galactose.

Assignees

• Univ South China Tech

Inventors

• Zheng Suiping
• Tang Shiming
• Zhang Zhiteng
• Liao Daocheng
• Lin Ying

Classifications

• C12P7/40 Primary

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C12R2001/19

  Escherichia coli · CPC title

• C12Y101/03041 Primary

  Alditol oxidase (1.1.3.41) · CPC title