Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Chitosanase gene, chitosanase as well as preparation method and application of chitosanase

A technology of chitosanase and gene, applied in the field of genetic engineering, can solve the problems of insufficient research on fungal chitosanase, achieve good stability and codon adaptability, good application prospects and industrial value, and improve sequence stability Effect

Pending Publication Date: 2020-11-06
深圳润康生态环境股份有限公司
View PDF8 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide polycanase gene csn and corresponding recombinant expression vector, recombinant expression bacterial strain, and chitosanase Csn and its preparation method and application, aim to solve existing research on fungal chitosanase in existing research Insufficient technical issues

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
  • Chitosanase gene, chitosanase as well as preparation method and application of chitosanase
  • Chitosanase gene, chitosanase as well as preparation method and application of chitosanase
  • Chitosanase gene, chitosanase as well as preparation method and application of chitosanase

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0045] Correspondingly, the embodiment of the present invention also provides a kind of preparation method of chitosanase Csn, it comprises the steps:

[0046] S1. Provide chitosanase gene csn, expression vector and expression strain;

[0047] S2, amplifying the chitosanase gene csn, and connecting the resulting amplified product with the expression vector to obtain a recombinant expression vector;

[0048] S3. Transforming the recombinant expression vector into an expression strain to obtain a recombinant expression strain;

[0049] S4. Cultivate the recombinant expression strain to obtain chitosanase Csn;

[0050] Wherein, the nucleotide sequence of the chitosanase gene csn is shown in SEQ ID NO:1.

[0051] In the preparation method of chitosanase Csn that the embodiment of the present invention provides, by amplifying chitosanase gene csn, obtain corresponding recombinant expression vector, recombinant expression bacterial strain, obtain chitosanase Csn through culture ag...

Embodiment 1

[0076] The present embodiment provides chitosanase CsnLr gene sequence analysis and codon optimization process, comprises the steps:

[0077] By consulting the glycoside hydrolase bioinformatics database (http: / / www.cazy.org / ), it is found that the chitosanase GH46 family has the most sequences and related information. At present, there are 852 chitosanases recorded in the GH46 family. Among them, 6 are from archaea, 784 are from bacteria, 60 are from viruses, and 2 are from fungi. According to the results of reviewing the literature, the chitosanases of the GH46 family of recombinant expression and characteristic analysis are all of bacterial origin, and chitosanases of fungal origin have not been reported yet. In view of the above analysis, the embodiment of the present invention expects to lay the foundation for excavating chitosanase derived from fungi of the GH46 family through recombinant expression of chitosanase derived from fungi.

[0078] By analyzing the glycoside ...

Embodiment 2

[0080] The present embodiment provides a kind of construction process of the recombinant expression vector containing chitosanase gene csn, specifically as follows:

[0081] (1) according to the sequence design of chitosanase gene csn a pair of primers CSN-F and CSN-R, its nucleotide sequence is as shown in SEQ ID NO:3-4, is used for amplifying chitosanase gene csn ;

[0082] (2) Obtain the chitosanase gene csn by PCR amplification, use the restriction endonuclease EcoRI and XbaI to digest the expression vector pPICZαA and chitosanase gene csn overnight respectively, and the expression vector pPICZαA and chitosan enzyme digested overnight The carbohydrase gene csn is purified and recovered for ligation reaction;

[0083] (3) Transfer the ligation reaction product into Escherichia coli Top10 by the heat shock method, and verify the recombinant transformant by bacterial liquid PCR;

[0084] (4) Transformants successfully verified were inserted into LBZ liquid medium, plasmids ...

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 belongs to the technical field of gene engineering, and particularly relates to a chitosanase gene, chitosanase as well as a preparation method and application of the chitosanase. A nucleotide sequence of the chitosanase gene csn is as shown in SEQ ID NO: 1, and the chitosanase gene csn is obtained by comprehensively optimizing the sequence of the chitosanase from lichtheimia ramosaaccording to the preference of pichia pastoris codons. An amino acid sequence of the chitosanase Csn is as shown in SEQ ID NO: 2. The chitosanase Csn is fungal chitosanase, not only enriches the variety and source ways of the chitosanase, but also has relatively high enzymatic activity, shows good stability in relatively wide temperature and pH ranges, and has good application prospects and industrial values.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to chitosanase gene csn, corresponding recombinant expression vector, recombinant expression strain, chitosanase Csn and its preparation method and application. Background technique [0002] As a biologically active substance, oligochitosan is composed of glucosamine and N-acetylglucosamine connected randomly through β-1,4 glycosidic bonds. Studies have shown that chitosan has antibacterial, anti-tumor, and immune-enhancing effects, so it is widely used in agriculture, animal husbandry, food, and medicine. At present, the preparation of oligochitosan is mainly divided into physical method, chemical method and enzymatic method. Compared with physical and chemical methods, enzymatic preparation of chitooligosaccharides has many advantages such as mild reaction conditions, complete product structure, easy control of the process, and no pollution to the environm...

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
IPC IPC(8): C12N15/56C12N9/24C12N15/81C12N1/19C12P19/26C12R1/84
CPCC12N9/2402C12N15/815C12P19/26C12Y302/01132C12N2800/22
Inventor 王建荣祝木金余思王平孟建军刘小慧曹革
Owner 深圳润康生态环境股份有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com