NZ744061B2 - Novel ?-galactosidase - Google Patents
Novel ?-galactosidaseInfo
- Publication number
- NZ744061B2 NZ744061B2 NZ744061A NZ74406116A NZ744061B2 NZ 744061 B2 NZ744061 B2 NZ 744061B2 NZ 744061 A NZ744061 A NZ 744061A NZ 74406116 A NZ74406116 A NZ 74406116A NZ 744061 B2 NZ744061 B2 NZ 744061B2
- Authority
- NZ
- New Zealand
- Prior art keywords
- enzyme
- galactosidase
- amino acid
- acid sequence
- seq
- Prior art date
Links
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
- C12N9/2471—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
Abstract
The purpose is to provide a novel ß-galactosidase useful in the production of oligosaccharides. Disclosed is a ß-galactosidase including an amino acid sequence of any of SEQ ID NOS: 1-4 or an amino acid sequence having 80% or greater identity with the amino acid sequences.
Claims (15)
1. A ß-galactosidase enzyme comprising an amino acid sequence of any one of 5 SEQ ID NOs: 1 to 4 or an amino acid sequence that is 80% or more identical to said amino acid sequence which possesses the following enzymological properties: (1) an enzymatic action by which the enzyme has a lactose hydrolyzing activity and a transgalactosylation activity, wherein the activity of the enzyme to transfer a galactosyl residue via ß-1,4-linkage is superior to that via ß-1,6-, ß-1,3-, or 10 ß-1,2-linkage; (2) an optimum temperature of 70°C; and (3) a molecular weight of about 104 kDa, about 64 kDa, or about 61 kDa (by
2. SDS-PAGE) for the enzyme without sugar chains. 15 2. The ß-galactosidase enzyme according to claim 1, wherein the amino acid sequence is an amino acid sequence that is 85% or more identical to the amino acid sequence of any one of SEQ ID NOs: 1 to 4.
3. The ß-galactosidase enzyme according to claim 1 or claim 2, wherein the 20 amino acid sequence consists of an amino acid sequence having a length equal to or less than that of the amino acid sequence of SEQ ID NO: 1.
4. The ß-galactosidase enzyme according to any one of claims 1 to 3, which further possesses the following enzymological properties: 25 (4) an optimum pH of 4 to 5; (5) a pH stability in which the enzyme is stable in a range of pH 2 to 8 (at 40°C for 30 minutes); and (6) a thermostability in which the enzyme is stable in a temperature range of 30°C to 60°C (at pH 6.0 for 30 minutes).
5. The ß-galactosidase enzyme according to any one of claims 1 to 3, which further possesses the following enzymological properties: (4) an optimum pH of 4 to 5; (5) a pH stability in which the enzyme is stable in a range of pH 2 to 9 (at 40°C 35 for 30 minutes); and (6) a thermostability in which the enzyme is stable in a temperature range of 30°C to 65°C (at pH 6.0 for 30 minutes).
6. An enzyme preparation comprising, as an active ingredient, the 5 ß-galactosidase enzyme according to any one of claims 1 to 5.
7. A ß-galactosidase gene consisting of a DNA selected from the group consisting (a) a DNA encoding the amino acid sequences of any one of SEQ ID NOs: 1 to 10 4; (b) a DNA consisting of the base sequence of any one of SEQ ID NOs: 5 to 8 and 16; and (c) a DNA comprising a base sequence equivalent to that of any one of SEQ ID NOs: 5 to 8 and 16 and encoding a protein with ß-galactosidase activity.
8. A recombinant DNA comprising the ß-galactosidase gene according to claim 7.
9. A microorganism carrying the recombinant DNA according to claim 8. 20 10. A method for producing a ß-galactosidase enzyme, comprising the steps of: (1) culturing cells of Cryptococcus terrestris; and (2) collecting the ß-galactosidase enzyme from the cultured medium and/or cells, wherein the Cryptococcus terrestris is Cryptococcus terrestris strain
10. MM13-F2171 or a mutant strain thereof.
11. A method for producing a ß-galactosidase enzyme, comprising the steps of: (i) culturing the microorganism of claim 9 under conditions allowing the production of protein encoded by the gene; and (ii) collecting the protein that has been produced.
12. A method for producing oligosaccharides, comprising a step of subjecting the ß-galactosidase enzyme according to any one of claims 1 to 5 to a reaction with a disaccharide, oligosaccharide, or polysaccharide having at least one of ß-1,3-, ß-1,4-, and ß-1,6-linkages.
13. A method for producing oligosaccharides, comprising a step of subjecting the ß-galactosidase enzyme according to any one of claims 1 to 5 to a reaction with lactose. 5
14. The method according to claim 12 or 13, wherein the reaction temperature in the step is from 30°C to 75°C.
15. Use of the ß-galactosidase enzyme according to any one of claims 1 to 5 for the production of oligosaccharides. DRAWINGS
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| NZ783737A NZ783737B2 (en) | 2016-12-27 | Novel b-galactosidase |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2015257705 | 2015-12-29 | ||
| PCT/JP2016/089001 WO2017115826A1 (en) | 2015-12-29 | 2016-12-27 | Novel β-galactosidase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| NZ744061A NZ744061A (en) | 2025-06-27 |
| NZ744061B2 true NZ744061B2 (en) | 2025-09-30 |
Family
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