# US 2018/0023073 A1 · Aldolase, aldolase mutant, and method and composition for producing tagatose by using same

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- Number: US 2018/0023073 A1
- Document ID: US20180023073A1
- Status: Published application
- Kind: A1
- Application number: 15/718,400
- Filed: 2017-09-28
- Published: 2018-01-25
- Earliest claimed: 2013-07-29
- Assignees: Samyang Corporation
- Applicants: Samyang Corporation
- Inventors: Deok-Kun OH; Seung-Hye Hong; Seon-Hwa Lee
- Web page: https://patentgrep.com/patent/US20180023073A1

## CPC

- C12N 9/88: Lyases (4.)
- C12P 7/26: Ketones
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12Y 401/02013: Fructose-bisphosphate aldolase (4.1.2.13)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES

## Abstract

There are provided aldolase, an aldolase mutant, a method for producing tagatose, and a composition for producing tagatose using the same. The technical feature of the present invention is environment-friendly due to the use of an enzyme acquired from microorganisms, requires only a simple process of enzyme-immobilization, uses a low-cost substrate compared with that of a conventional method for producing tagatose, and has a remarkably high yield, thereby greatly reducing production cost while maximizing production effect.

## Claims

### Claim 1

A method of producing tagatose from fructose, the method comprising:
reacting fructose 6-phosphate with tagatose 6-phosphate epimerase or a mutant thereof to obtain tagatose 6-phosphate; and
converting the tagatose 6-phosphate to tagatose.

### Claim 2 (depends on 1)

The method of claim 1, wherein the tagatose 6-phosphate epimerase catalyzes the conversion of fructose 6-phosphate to tagatose-6-phosphate.

### Claim 3 (depends on 1)

The method of claim 1, wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 1.

### Claim 4 (depends on 1)

The method of claim 1, wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 2.

### Claim 5 (depends on 1)

The method of claim 1, wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 3.

### Claim 6 (depends on 1)

The method of claim 1, wherein the tagatose 6-phosphate epimerase is SEQ ID NOS: 4.

### Claim 7 (depends on 1)

The method of claim 1, wherein the mutant comprises at least one amino acid substitution of SEQ ID NO: 1 selected from the group consisting of:
i) a substitution of arginine residue with glutamine at a position corresponding to position 332,
ii) a substitution of glutamine residue with alanine at a position corresponding to position 314,
iii) a substitution of histidine residue with alanine at a position corresponding to position 227, and
iv) a substitution of serine residue with alanine at a position corresponding to position 62.

### Claim 8 (depends on 1)

The method of claim 1, wherein the fructose 6-phosphate is obtained by treating fructose or a fructose-containing material with hexokinase.

### Claim 9 (depends on 1)

The method of claim 1, wherein the tagatose is obtained by treating the tagatose 6-phosphate with phytase.

### Claim 10 (depends on 1)

The method of claim 1, wherein the reaction of the fructose 6-phosphate with the tagatose 6-phosphate epimerase is performed at a pH from 7.0 to 9.0.

### Claim 11 (depends on 1)

The method of claim 1, wherein the reaction of the fructose 6-phosphate with the tagatose 6-phosphate epimerase is performed at a temperature from 30 ° C. to 70 ° C.