Construction method of xylanase hybrid enzyme engineering strain

A technology of xylanase and engineering strains, applied in the biological field to achieve the effect of increasing the optimum temperature

Active Publication Date: 2016-04-20
XINXIANG MEDICAL UNIV
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are relatively few studies on improving the stability of xylanase through glycosylation modification, so there is some room for exploration

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Construction method of xylanase hybrid enzyme engineering strain
  • Construction method of xylanase hybrid enzyme engineering strain
  • Construction method of xylanase hybrid enzyme engineering strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0023] 1 Materials and methods

[0024] 1.1 Strains, plasmids and reagents

[0025] Aspergillus niger (Aspergillus niger) XZ-3S, Escherichia coli (Escherichiacoli) JM109, DH5α were preserved by the author's laboratory; eukaryotic expression plasmid pPIC9K and Pichia pastoris (Pichia pastoris) GS115 were purchased from Invitrogen. DNA polymerase, restriction endonuclease and ligase were purchased from TaKaRa Company; ampicillin (Amp) was purchased from Sangon Company; gel recovery kit and plasmid extraction kit were purchased from Sangon Company; Ethidium bromide was purchased from Amresco Company; yeast powder, peptone and agarose were purchased from BBI Company; G418 and amino-free yeast nitrogen source (YNB) were purchased from Amresco Company; birch xylan was purchased from Sigma Company; other biochemical reagents were do...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a construction method of a xylanase hybrid enzyme engineering strain. The method comprises the steps that homologies of a xylanase XynZF-2 gene sequence and a heat-resistant xylanase EvXyn11 gene sequence are compared through bioinformatics, 48 amino acids of a XynZF-2 N-terminal is replaced with 34 amino acids of a xylanase EvXyn11 N-terminal through N-terminal replacement, aromatic amino acids P9Y and H14F are introduced, disulfide bonds Cys38-Cys191 are continuously introduced into the XynZF-2 enzyme C-terminal, hydrophobic amino acids K164M, G166A, N160I, V111A and G109A are introduced into alpha-spiral and cord areas, glycosylation modification sites E42N and F17S are introduced finally, multiple locus mutational hybrid enzyme XynZL is constructed, and the heat stability of the hybrid enzyme XynZL is expected to be improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for constructing a xylanase hybrid enzyme engineering strain. Background technique [0002] Xylanase (EC3.2.1.8) is a general term for a class of multifunctional enzymes that catalyze the hydrolysis of xylan. Xylanase is widely distributed in nature, such as animals, plants and microorganisms, and has a wide variety of applications. At present, there are many reports on xylanases derived from microorganisms, and the most studied are xylanases derived from bacteria and fungi. Among them, bacteria can produce alkaline and acidic xylanases, while fungi can produce alkaline xylanases. At present, xylanase is mainly produced by fermentation of microorganisms such as fungi and bacteria. [0003] With the continuous development of biotechnology, the research and development of xylanase has attracted much attention. In terms of industrial application, xylanase has great potential ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/56C12N15/81C12R1/84
CPCC12N9/2482C12Y302/01008
Inventor 周晨妍刘振华王燕王拥涛李同彪朱新术
Owner XINXIANG MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap