Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

A kind of chitin deacetylase and its application

A kind of technology of element deacetylase and element deacetylase, applied in the field of chitin deacetylase, can solve the problems of optimal temperature difference, unsuitable reaction conditions of industrial materials, etc., and achieve the effect of low production conditions

Active Publication Date: 2022-06-28
JILIN COFCO BIOCHEM +2
View PDF12 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The CDA derived from fungi found so far are basically single-peptide chain glycoproteins, showing good thermal stability, but the position, optimum pH value, optimum temperature, molecular weight, isoelectricity, etc. point and the reaction to metal ions and acetic acid are quite different
In addition, the optimum pH of the CDA (hereinafter also referred to as CliCDA) derived from the reported plant pathogenic bacterium Bean anthracnose is a slightly alkaline environment, which is not suitable for the reaction conditions of industrial materials (slightly acidic)
[0006] In summary, the activity of CDA found so far (especially the activity under acidic conditions) still cannot fully meet the actual needs of industrialized production of chitosan, therefore, screening highly active CDA is still a need in industrialized applications. important issues to solve

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
  • A kind of chitin deacetylase and its application
  • A kind of chitin deacetylase and its application
  • A kind of chitin deacetylase and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0043] According to a preferred embodiment of the present invention, Escherichia coli transformed cells or Bacillus subtilis transformed cells can be obtained by the following method: by chemical transformation, the expression vector of the present invention is transformed into Escherichia coli competent cells or Bacillus subtilis competent cells cells; the bacterial suspension was then spread on a plate and incubated until a single colony appeared.

[0044] Five. The kit of the present invention

[0045] The kit of the present invention is a kit comprising one or more of the chitin deacetylase of the present invention, a DNA molecule encoding the chitin deacetylase of the present invention, the expression vector of the present invention, and the transformed cell of the present invention .

[0046] The kit may comprise a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, and the lik...

Embodiment 1

[0058] Example 1 Design and screening of chitin deacetylase

[0059] A potential thermostable sugar isomerase was searched in the NCBI database, and an unknown protein from Verticillium rape Vlo was found through sequence alignment analysis and strain source investigation, and its amino acid sequence is as shown in SEQ ID NO: 1 shown, it is presumed to have deacetylase activity.

[0060] Furthermore, the coding sequence of the amino acid sequence was codon-optimized to obtain the nucleotide sequence shown in SEQ ID NO:2.

Embodiment 2

[0061] Example 2 Expression of chitin deacetylase in Escherichia coli

[0062] The nucleotide sequence shown in SEQ ID NO: 2 was synthesized by General Biosystems (Anhui) Co., Ltd., a BamH I restriction site was added to its 5' end, and a hexahistidine tag was added before the stop codon. coding sequence, and Pst I and Not I restriction sites were added at the 3' end. The synthesized fragment was double-digested by BamH I and Not I (New England Biolabs, NEB), and the double-digested fragment was ligated with T4 DNA ligase (Takara) to the same double-digested by BamHI and NotI. pET24a(+) vector (General Biosystems (Anhui) Co., Ltd.). The ligated products were transformed into E. coli DH5α competent cells (Beijing Quanshijin Biotechnology Co., Ltd.), cultured on LB solid plates with kanamycin, and positive clones were screened. A single colony was picked for colony PCR verification; clones that were positive by colony PCR were verified by sequencing.

[0063] Plasmid extracti...

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 present invention screens out several unknown proteins by analyzing and researching different genome sequences of different species, and further identifies a chitin deacetylase with excellent deacetylase activity. More specifically, the present invention relates to a A chitin deacetylase derived from Verticillium longisporum Vlo. Compared with the reported chitin deacetylase, the chitin deacetylase requires less production conditions and has higher biosafety properties, and has characteristics more suitable for industrial applications, showing a wide range of application potential.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a chitin deacetylase derived from Verticillium longisporum VL1 and its application. Background technique [0002] Chitin, also known as chitin, is a linear polymer aminopolysaccharide composed of N-acetylamino-D-glucose monomers (D-GlcNAc) linked by β-1,4 glycosidic bonds, mainly found in invertebrates. (especially the shells of crustaceans such as shrimps, crabs, insects, etc.), seaweeds (such as green algae), fungi (such as molds), etc., it is another important type of polysaccharide other than cellulose. However, chitin is insoluble in water, dilute acid, alkali, ethanol or other organic solvents, which greatly limits its utilization value, and the product chitosan after deacetylation depends on the degree of deacetylation and can be dissolved in acid. and neutral aqueous solutions. Because chitosan has excellent properties such as biofunctionality and compatibil...

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 Patents(China)
IPC IPC(8): C12N9/80C12N15/55C12N15/70C12N15/75C12N1/21C12P19/26C12R1/125C12R1/19
CPCC12N9/80C12N15/70C12N15/75C12P19/26C12Y305/01041
Inventor 佟毅沈雪梅王靖张媛刘颖慰王小艳陈博彭超李义周勇卢宗梅满云
Owner JILIN COFCO BIOCHEM
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
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