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

A method for producing candida antarctica lipase b and the specific dna molecule used therefor

A technology of Candida Antarctica and DNA molecules, applied in the biological field, can solve the problems of high cost of eukaryotic expression and difficult promotion of industrial application.

Active Publication Date: 2022-04-12
INST OF ZOOLOGY CHINESE ACAD OF SCI
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cost of eukaryotic expression is high, and industrial application is difficult to promote, so low-cost prokaryotic expression has become a research trend

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 method for producing candida antarctica lipase b and the specific dna molecule used therefor
  • A method for producing candida antarctica lipase b and the specific dna molecule used therefor
  • A method for producing candida antarctica lipase b and the specific dna molecule used therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1, adopt CALB fusion gene to produce Candida antarctica lipase B

[0068] 1. Construction of recombinant plasmids

[0069] 1. Optimize the codon according to the amino acid sequence of the CALB protein, and artificially synthesize the CALB fusion gene with 6 His tags at the N-terminal.

[0070] The nucleotide sequence of the CALB fusion gene is shown in SEQ ID No:3.

[0071] The CALB fusion gene encodes the CALB fusion protein shown in SEQ ID No:4.

[0072] 2. Construction of recombinant plasmid pET30a-CALB

[0073] The small DNA fragment between the restriction endonucleases NdeI and HindIII of the plasmid pET30a was replaced with the DNA molecule shown in the 4th to 978th ​​positions from the 5' end of SEQ ID NO: 3, and the other sequences were unchanged to obtain the recombinant plasmid pET30a -CALB.

[0074] The nucleotide sequence of the recombinant plasmid pET30a-CALB is shown in SEQ ID NO:8.

[0075] The recombinant plasmid pET30a-CALB expresses t...

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 method for producing Candida antarctica lipase B and the specific DNA molecule used therefor. The nucleotide sequence of the specific DNA molecule is shown in SEQ ID NO:3. Experiments have shown that the recombinant plasmid pET30a‑CALB prokaryotically expresses CALB inclusion bodies, and then undergoes renaturation to obtain high-purity (over 95%) and high-activity CALB protein; the enzyme activity of CALB protein obtained from 1L of culture medium is about 21252U, and the specific activity About 253U / mg; much higher than the specific activity of the CALB protein prepared in the existing literature. It can be seen that the present invention significantly improves the output of CALB, and the production process is simple, and the completion can be applied to actual production. The invention has important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for producing Candida antarctica lipase B and the specific DNA molecule used therein. Background technique [0002] Candida antarctica Lipase B (CALB) is a lipase extracted from Candida antarctica samples, which can catalyze the synthesis of biological organic esters (such as surfactants, pharmaceutical intermediates, etc.) , hydrolysis of esters, transesterification and other types of reactions, it has strong catalytic activity, high stereoselectivity and high stability for water-soluble and water-insoluble substances, and has become one of the most widely used lipases in the world. The low-cost mass production will make it have a broader application prospect in the field of green biochemical medicine. [0003] In 1994, Uppenberg J. determined the amino acid sequence and its three-dimensional structure of CALB (UppenbergJ., Patkar S., Bergfors T. and Jones T.A....

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): C12N15/55C12N15/70C12N1/21C07K19/00C12N9/20C12R1/19
CPCC12N9/20C12Y301/01003C12N15/70C07K2319/21
Inventor 江红邹振
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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