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A kind of β-glucosidase with galacto-oligosaccharide synthesis ability and its expression strain and application

A technology of glucosidase and galactooligosaccharides, which is applied in the field of β-glucosidase and its expression strains, can solve the problem of less β-glucosidase and achieve good stability and high transglycosidic activity

Active Publication Date: 2022-06-07
ANHUI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few GH1 family β-glucosidases currently used in the synthesis of galactooligosaccharides

Method used

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  • A kind of β-glucosidase with galacto-oligosaccharide synthesis ability and its expression strain and application
  • A kind of β-glucosidase with galacto-oligosaccharide synthesis ability and its expression strain and application
  • A kind of β-glucosidase with galacto-oligosaccharide synthesis ability and its expression strain and application

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Embodiment Construction

[0040] The implementation methods in the following examples are conventional methods unless otherwise specified.

[0041] (1) Construction of an expression strain containing the β-glucosidase gene with galacto-oligosaccharide synthesis ability derived from marine bacteria of the present invention

[0042] 1. Screening and identification of positive clones containing the β-glucosidase gene with galactooligosaccharide synthesis ability derived from marine bacteria of the present invention

[0043] From the shallow sea area near Zhaoshu Island in the South China Sea: water temperature 30℃, pH 7.9, water depth 4-5 meters (16°58′N, 112°16′E) seabed sediments, isolate microorganisms and extract genomes, take 1g of seabed sediments , and 50mL TYS liquid medium, 30 ℃, 120rpm, activated for 2h. Dilute the supernatant at different times and spread it on a TYS solid plate containing 0.1% (w / v) escin and 0.25% (w / v) ferric ammonium citrate, and invert at 16°C, 28°C, and 37°C for culture. ...

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Abstract

The present invention discloses a β-glucosidase capable of synthesizing galactooligosaccharides and its expression strain and application, wherein the β-glucosidase capable of synthesizing galactooligosaccharides is named BglD1, which originally originated from China The seabed sediments of the Xisha Islands in the South China Sea have one of the following amino acid sequences: (1) SEQ ID No: 2 in the sequence listing; (2) SEQ ID No: 2 in the sequence listing is substituted, deleted or added with one or more amino groups residues and encode the amino acid sequence of the same functional protein. The β-glucosidase with the ability to synthesize galactooligosaccharides of the present invention has β-glucosidase and β-galactosidase activities, and can synthesize galactooligosaccharides through transglycosidation, and the bond type of the product is mainly β(1→ 3) and β(1→4) glycosidic bonds.

Description

technical field [0001] The invention relates to a beta-glucosidase with galacto-oligosaccharide synthesis ability, an expression strain and application thereof, and belongs to the field of biotechnology. Background technique [0002] β-glucosidase (EC 3.1.2.21, β-glucosidase) belongs to the class of hydrolases, mainly hydrolyzes the β-1,4-glycosidic bond in glycosides or oligosaccharides, while releasing terminal non-reducing glucose or other aglycones. body. According to the similarity of amino acid sequence and spatial structure, it is classified into glycoside hydrolase 1 family (GH1), 3 family, 5 family, 9 family, 30 family and 116 family (http: / / www.cazy.org / ). GH1 family β-glucosidases have broad substrate catalytic properties and can have both β-galactosidase, β-xylase, β-mannase and other enzyme activities, so they have important industrial application value. [0003] Galacto-oligosaccharides (GOS) is an important prebiotic and functional oligosaccharide with natur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12N15/70C12N1/21C12P19/00C12P19/04C12P19/18C12R1/19
CPCY02E50/10
Inventor 房伟肖亚中邓鹏军方泽民刘娟娟
Owner ANHUI UNIVERSITY
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