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Beta-xylosidase mutant and use thereof

A mutant, glucosidase technology, applied in the field of enzymology, can solve the problems of strong randomness, large workload, poor enzymology properties, etc., and achieve the effect of wide application prospects

Active Publication Date: 2014-02-26
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The breeding of microbial strains is one of the most commonly used and simplest methods in industrial production and academic research, but it is often difficult to screen ideal strains due to heavy workload and strong randomness
Protein engineering is an emerging method that utilizes molecular biology and bioinformatics methods to conduct directed mutation and rational transformation of proteins to obtain ideal proteins or enzymes. Its advantages lie in relatively small workload and high probability of success, but Enzymatic properties of most mutant proteins were unchanged or worse

Method used

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  • Beta-xylosidase mutant and use thereof
  • Beta-xylosidase mutant and use thereof
  • Beta-xylosidase mutant and use thereof

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Embodiment 1

[0026] The preparation of embodiment 1β-xylosidase mutant

[0027] Materials: Escherichia coli strain XL10-gold and Escherichia coli M15 were purchased from Stratagene; Ni-NTA protein histidine purification medium was purchased from Intrivogen; restriction enzymes and modification enzymes were purchased from TaKaRa and MBI ; Xylooligosaccharides and xylose were purchased from Sigma; the rest of the reagents were conventional reagents purchased from the market unless otherwise specified.

[0028] 1) Cloning of β-xylosidase gene Exyl

[0029] A gene sequence encoding a putative glycosyl hydrolase (YP_003612882.1) of Enterobacter cloacae subsp.cloacae ATCC13047 was found from the GenBank gene database. Design the upstream primer Exyl-1: 5'-CTGAGATCTACTGCAATCTATAAGGACGCGGGAC-3' (containing a BglII restriction site) and the downstream primer Exyl-25'-ACTAAGCTTCTACGCGTGCTGAACCTGACAGGTG-3' (containing a HindIII restriction site), through the polymerase chain The β-xylosidase gene E...

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Abstract

The invention discloses a beta-xylosidase mutant and a use thereof. The beta-xylosidase mutant has an amino acid sequence as shown in SEQ ID NO:1. The mutant enzyme has high xylose and glucose tolerance compared with non-mutated beta-xylosidase, and has wide use in degradation of xylo-oligosaccharide.

Description

technical field [0001] The invention belongs to the field of enzymology, relates to a beta-xylosidase mutant, and also relates to the use of the mutant in degrading xylooligosaccharides. Background technique [0002] Xylan is the main component of hemicellulose, accounting for 30-35% of the total dry weight of hemicellulose. It is widely distributed in the cell walls of higher plants. It is a renewable resource with a huge stock in nature, and it is also easier to extract, A class of hemicelluloses that are degraded and utilized. Xylanase is a kind of complex enzyme system that can hydrolyze xylan to generate xylose, mainly composed of endoxylanase (EC.3.2.1.8) and β-xylosidase (EC3.2.1.37) (Beg Q.K., Kapoor M., Mahajan L., et al. 2001, Microbial xylanase and their industrial applications: A review. Appl. Microbiol Biotechnol., 56:326-338.). [0003] β-Xylosidase (β-D-xylosidase) is an exohydrolase, which cooperates with endoxylanase to degrade the degradation product of e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N15/56C12N1/15C12N1/19C12N1/21C12P19/14
CPCC12N9/2434C12Y302/01037
Inventor 杜丽琴黄金群王子龙王金佩黄日波张志凯韦宇拓
Owner GUANGXI UNIV
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