NZ783737B2 - Novel b-galactosidase - Google Patents
Novel b-galactosidaseInfo
- Publication number
- NZ783737B2 NZ783737B2 NZ783737A NZ78373716A NZ783737B2 NZ 783737 B2 NZ783737 B2 NZ 783737B2 NZ 783737 A NZ783737 A NZ 783737A NZ 78373716 A NZ78373716 A NZ 78373716A NZ 783737 B2 NZ783737 B2 NZ 783737B2
- Authority
- NZ
- New Zealand
- Prior art keywords
- enzyme
- galactosidase
- amino acid
- galactosidase enzyme
- acid sequence
- 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 present invention has a purpose of providing a novel ß-galactosidase enzyme useful for the production of oligosaccharides. Disclosed is a ß-galactosidase enzyme comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 4 or an amino acid sequence that is 80% or more identical to said amino acid sequence
Claims (17)
1. A ß-galactosidase enzyme comprising an amino acid sequence that is 90% or 5 more identical to any one of SEQ ID NOs: 1 to 4.
2. The ß-galactosidase enzyme according to claim 1, wherein the 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.
3. The ß-galactosidase enzyme according to claim 1 or claim 2, wherein the amino acid sequence comprises the amino acid sequence of any one of SEQ ID NOs: 1 to 4.
4. A ß-galactosidase enzyme according to claim 1 which possesses the following 15 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 ß-1,2- linkage; 20 (2) an optimum temperature of 70°C; and (3) a molecular weight of about 104 kDa, about 64 kDa, or about 61 kDa (by SDS-PAGE) for the enzyme without sugar chains.
5. The ß-galactosidase enzyme according to claim 4, which further possesses the 25 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 8 (at 40°C for 30 minutes); and (6) a thermostability in which the enzyme is stable in a temperature range of 30 30°C to 60°C (at pH 6.0 for 30 minutes).
6. The ß-galactosidase enzyme according to claim 4, which further possesses the following enzymological properties: (4) an optimum pH of 4 to 5; 35 (5) a pH stability in which the enzyme is stable in a range of pH 2 to 9 (at 40°C 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). 5
7. An enzyme preparation comprising, as an active ingredient, the ß-galactosidase enzyme according to any one of claims 1 to 6.
8. A ß-galactosidase gene consisting of a DNA selected from the group consisting 10 (a) a DNA encoding the amino acid sequences of any one of SEQ ID NOs: 1 to (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 that is 90% or more identical to that of 15 any one of SEQ ID NOs: 5 to 8 and 16 and encoding a protein with ß-galactosidase activity.
9. A recombinant DNA comprising the ß-galactosidase gene according to claim 8. 20
10. A microorganism carrying the recombinant DNA according to claim 9.
11. A method for producing a ß-galactosidase enzyme according to any one of claims 1 to 6, comprising the steps of: (1) culturing cells of Cryptococcus terrestris; and 25 (2) collecting the ß-galactosidase enzyme from the cultured medium and/or cells.
12. The method according to claim 11, wherein the Cryptococcus terrestris is Cryptococcus terrestris strain MM13-F2171 or a mutant strain thereof.
13. A method for producing a ß-galactosidase enzyme, comprising the steps of: (i) culturing the microorganism of claim 10 under conditions allowing the production of protein encoded by the gene; and (ii) collecting the protein that has been produced.
14. A method for producing oligosaccharides, comprising a step of subjecting the ß-galactosidase enzyme according to any one of claims 1 to 6 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.
15. A method for producing oligosaccharides, comprising a step of subjecting the ß-galactosidase enzyme according to any one of claims 1 to 6 to a reaction with lactose.
16. The method according to claim 14 or 15, wherein the reaction temperature in 10 the step is from 30°C to 75°C.
17. Use of the ß-galactosidase enzyme according to any one of claims 1 to 6 for the production of oligosaccharides. DRAWINGS
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2015257705 | 2015-12-29 | ||
| NZ744061A NZ744061B2 (en) | 2016-12-27 | Novel ?-galactosidase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| NZ783737A NZ783737A (en) | 2025-09-26 |
| NZ783737B2 true NZ783737B2 (en) | 2026-01-06 |
Family
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