Beta-glucosidase with galactooligosaccharide synthesis capability and expression strain and application thereof

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: 2019-06-14
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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  • Beta-glucosidase with galactooligosaccharide synthesis capability and expression strain and application thereof
  • Beta-glucosidase with galactooligosaccharide synthesis capability and expression strain and application thereof
  • Beta-glucosidase with galactooligosaccharide synthesis capability and expression strain and application thereof

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Embodiment Construction

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

[0041] (1) Containing the construction of the expression strain of the β-glucosidase gene with the ability to synthesize galactooligosaccharides from marine bacteria of the present invention

[0042] 1. Screening and identification of positive clones containing the β-glucosidase gene with galacto-oligosaccharide 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°C, pH 7.9, water depth 4-5 meters (16°58′N, 112°16′E) seabed sediment, isolate microorganisms and extract genome, take 1g of seabed sediment , with 50mL TYS liquid medium, 30°C, 120rpm, activated for 2h. The supernatant was diluted in different multiples and spread on TYS solid plates containing 0.1% (w / v) aescin and 0.25% (w / v) ferric ammonium citrate, and incubated upside down at 16°C, ...

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Abstract

The invention discloses beta-glucosidase with galactooligosaccharide synthesis capability and an expression strain and application thereof, wherein the beta-glucosidase with the galactooligosaccharidesynthesis capability is named BglD1, derived from seabed sediments of the Xisha Islands of the South China Sea, with one of the following amino acid sequences: (1) SEQ ID No: 2 in the sequence listing; (2) the amino acid sequence obtained by substitution, deletion or addition of one or a plurality of amino residues of SEQ ID No: 2 in the sequence listing and encoding the same functional protein.The beta-glucosidase with the galactooligosaccharide synthesis capability has beta-glucosidase and beta-galactosidase activity, and can synthesize galactooligosaccharide by transglycosylation. The product bond type is mainly beta (1->3) and beta (1->4) glycosidic bonds.

Description

technical field [0001] The invention relates to a beta-glucosidase capable of synthesizing galacto-oligosaccharides and its expression strain and application, belonging to the field of biotechnology. Background technique [0002] β-glucosidase (EC 3.1.2.21, β-glucosidase) belongs to hydrolase, which mainly hydrolyzes β-1,4-glucosidic bonds in glycosides or oligosaccharides, and at the same time releases terminal non-reducing glucose or other aglycosides. body. According to the similarity of amino acid sequence and spatial structure, it is classified into glycoside hydrolase family 1 (GH1), family 3, family 5, family 9, family 30 and family 116 (http: / / www.cazy.org / ). The GH1 family β-glucosidase has broad substrate catalytic properties, and can have enzyme activities such as β-galactosidase, β-xylosidase, and β-mannase, so it has important industrial application value. [0003] Galacto-oligosaccharides (GOS) is an important prebiotic, a functional oligosaccharide with natu...

Claims

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Application Information

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Patent Type & Authority Applications(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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