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Applications of beta-glucosidase SPBGL5 in hydrolysis of xylan polysaccharide substances

A technology of glucosidase and polysaccharides, applied in the field of enzymatic engineering, to achieve the effect of great application potential

Active Publication Date: 2018-02-16
GUANGXI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] However, there is no patent for β-glucosidase that can act on beech xylan, birch wood xylan, oat xylan, corncob xylan and wheat arabinoxylan alone.

Method used

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  • Applications of beta-glucosidase SPBGL5 in hydrolysis of xylan polysaccharide substances
  • Applications of beta-glucosidase SPBGL5 in hydrolysis of xylan polysaccharide substances
  • Applications of beta-glucosidase SPBGL5 in hydrolysis of xylan polysaccharide substances

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Embodiment 1

[0032] 1. Cloning of β-glucosidase gene spbgl5

[0033] The Sphingomonas strain numbered ATCC 31461 was purchased from the American Type Culture Collection, and the total DNA of the bacteria was extracted. The nucleotide sequence of the gene encoding β-glucosidase was obtained from the NCBI gene database, the gene sequence number is NZ_AGFU01000034, which is located at 124633...127032 of the genome sequence of the Sphingomonas strain. VectorNTI and ORF finder were used to analyze the restriction site and ORF of the gene, and SMART and SignalP 4.1 were used to analyze the structural domain and predict the signal peptide of the protein encoded by the gene.

[0034] According to the analysis results, remove the signal peptide, use the selected restriction enzyme site, and add 6 His tags (underlined part) at the C-terminus, design a pair of PCR primers to amplify the target fragment, and the primer sequences are as follows:

[0035] 5-F: 5'-TCCTCATGAAACAGCCTGCCGCAGCGCAGACCGCC-3';...

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Abstract

The invention belongs to the technical field of enzymology engineering, and particularly relates to applications of beta-glucosidase SPBGL5 in hydrolysis of xylan polysaccharide substances. Accordingto the present invention, a recombinant Escherichia coli strain containing a beta-glucosidase gene spbgl5 from Sphingomonas sp. ATCC 31461 is constructed, wherein the gene is subjected to induced expression and nickel affinity chromatography purification in Escherichia coli to obtain an expression product beta-glucosidase SPBGL5; and the obtained beta-glucosidase SPBGL5 has the activities of beta-glucosidase, further has the activities of beta-xylosidase and beta-arabinosidase, can separately hydrolyze beech xylan, birch xylan, oat xylan, corncob xylan, wheat araboxylan and lichenin, and has great application potential in degradation of various xylan polysaccharide substances.

Description

technical field [0001] The invention belongs to the technical field of enzymatic engineering, and specifically relates to the application of a β-glucosidase SPBGL5 in hydrolyzing xylan polysaccharides. Background technique [0002] β-glucosidase (beta-glucosidase, EC 3.2.1.21), also known as cellobiase, gentiobiase and amygdalinase, broadly refers to a class that can catalyze the transfer of sugar groups between oxygen nucleophiles Enzyme (Singhania R R, Patel A K, Sukumaran R K, et al.Role and aignificance of beta-glucosidases in the hydrolysis of cellulose for bioethanol production[J].Bioresour Technol,2003,127:500-507.), is a cellulase system important constituents. [0003] β-glucosidase exists widely in nature, and the structure and catalytic properties of β-glucosidase from different sources are also different. Sources in plants include almonds, soybeans, ginseng, etc.; sources in animals include honey and pig liver, etc. -22.), the sources of microorganisms include...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12P19/14C12P19/02
CPCC12N9/2445C12P19/02C12P19/14C12Y302/01021
Inventor 庞浩唐婷婷杜丽琴汤宏赤黄日波林丽华郭媛严少敏
Owner GUANGXI ACAD OF SCI
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