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

Self-aggregation short-peptide induction based immobilized nitrilase and preparation method and application thereof

A nitrilase, self-aggregation technology, applied in the directions of hydrolase, immobilized on/in organic carrier, chemical industry, etc., to achieve the effects of good substrate tolerance, simple immobilization operation, and easy preservation

Inactive Publication Date: 2014-04-30
SOUTH CHINA UNIV OF TECH +1
View PDF4 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to overcome the shortcoming of existing nitrilase immobilization method, provide a kind of nitrilase immobilization method based on self-aggregation short peptide induced aggregation

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
  • Self-aggregation short-peptide induction based immobilized nitrilase and preparation method and application thereof
  • Self-aggregation short-peptide induction based immobilized nitrilase and preparation method and application thereof
  • Self-aggregation short-peptide induction based immobilized nitrilase and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1. Construction of expression vectors containing coding short peptides

[0041] According to the amino acid sequence of PT-linker and 18A, the corresponding DNA sequence PT-linker+18A was synthesized in a gene synthesis company, and a Hind III enzyme cleavage site was introduced upstream of the PT-linker+18A DNA sequence, and an Xho I enzyme site was introduced downstream . Using the plasmid pET-30a(+) as the basic plasmid, digest it with Hind III and XhoI, connect it into the corresponding digested PT-linker+18A sequence, transform it into E.coli DH5α competent cells, and perform colony PCR and enzyme digestion And sequencing analysis identified positive transformants to obtain plasmid pET-aIB, such as figure 1 shown.

[0042] 2. Construction of engineering bacteria expressing nitrilase active aggregates in Escherichia coli cells

[0043] Using the genome of Alcaligenes faecalis JM3 as a template, a 1 kb nitrilase gene fragment was amplified by PCR under the action ...

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a self-aggregation short-peptide induction based immobilized nitrilase and a preparation method and application thereof. The preparation method comprises the following steps: (1) splicing connecting peptides and self-aggregation short peptides, so as to construct an expression vector; (2) connecting a nitrilase gene and the expression vector, and transforming into a recipient strain, so as to obtain an engineering strain; (3) carrying out induced expression on the engineering strain, carrying out wall breaking treatment on cells, and centrifuging and / or filtrating to obtain a precipitate, namely a nitrilase endoenzyme aggregate; (4) immobilizing the obtained nitrilase endoenzyme aggregate in alginate, thereby obtaining the immobilized nitrilase. According to the immobilized nitrilase aggregate obtained by the method, the particle diameter of the immobilized nitrilase is about 1mm. According to the immobilized nitrilase, particles can be subjected to freeze-drying preservation, and the enzyme activity is reserved to 90% when the immobilized nitrilase is preserved for 20 days at the temperature of 37 DEG C; the activity of the immobilized nitrilase is high, and the operating flow is simple.

Description

technical field [0001] The invention belongs to the fields of bioengineering and biocatalysis. The invention relates to a method for aggregation, expression and immobilization of nitrilase induced by self-aggregation short peptides. More specifically, the present invention constructs a nitrilase genetically engineered bacterium expressed in the form of enzyme aggregates in vivo, the aggregates with nitrilase activity are fixed in sodium alginate solution, and the racemization is catalyzed in aqueous solution The nitriles are converted to the corresponding carboxylic acids. Background technique [0002] Nitrilase can convert nitriles into corresponding carboxylic acids and ammonia through a one-step catalytic reaction in aqueous solution. In recent years, many experts and scholars have used nitrilase to catalyze the biotransformation process of fatty nitriles, aromatic nitriles and alicyclic nitriles into corresponding organic carboxylic acids, and there have been very succ...

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): C12N15/70C12N9/78C12N11/10C12P7/42
CPCY02P20/50
Inventor 李爽杨晓锋林章凛王菊芳王小宁
Owner SOUTH CHINA UNIV OF TECH
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