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

Recombined yeast strain expressing cellulase and applications thereof

A technology of cellulase and yeast strains, applied in the field of genetic engineering, can solve the problems of inability to meet large-scale industrial production, lack of processing and modification functions of bacteria, low enzyme production and activity, etc., and achieve improved experimental conditions, strong tolerance, The effect of optimization of experimental conditions

Inactive Publication Date: 2012-08-22
陈战
View PDF5 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the prokaryotic cell expression system based on bacteria was first used in the production of cellulase, and its disadvantage is that the enzyme activity is not high, which is related to the lack of post-translational processing and modification functions of bacteria.
The production of cellulase by eukaryotic cell expression systems based on Saccharomyces cerevisiae and Pichia pastoris has attracted more and more attention, but the fermentation level is low, generally lower than 1g / L, which still cannot meet the requirements of large-scale industrial production. need
The cellulase production strains cultivated at present generally have the disadvantage of easy degradation, need to be cultivated repeatedly, and the enzyme yield and activity are not high, the enzyme activity of the produced strains is mostly lower than 300U / mL, and the yeast fermentation level of the enzyme is lower than 1 g / L

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
  • Recombined yeast strain expressing cellulase and applications thereof
  • Recombined yeast strain expressing cellulase and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] As shown in the accompanying drawings of the description, the cDNA gene sequence of Trichoderma reesei endonuclease EGI is from the 5' end to the 3' end, and the cDNA of Trichoderma reesei endonuclease containing the EcoRI-XbaI restriction site obtained by PCR amplification fragments, insert figure 1 The multiple cloning site MCS2 of the plasmid pPICZαABC3 was constructed, and a multi-copy system was constructed. Sequencing confirmed that the cDNA gene expression vector 4 of Trichoderma reesei endonuclease had been successfully constructed. This expression vector 4 was integrated into Pichia pastoris GS115. After screening on YPD plate, transfer to YPD liquid medium for culture. After the yeast is harvested, the enzyme protein is purified, and a high-yield strain with high enzyme specific activity and high yield is obtained by screening, that is, the recombinant Pichia pastoris 5-reesei endonuclease EGI strain expressing cellulase.

Embodiment 2

[0035] As shown in the accompanying drawings of the description, the cDNA gene sequence of Trichoderma reesei endonuclease EGII is from the 5' end to the 3' end, and the cDNA of Trichoderma reesei endonuclease containing the EcoRI-XbaI restriction site obtained by PCR amplification fragments, insert figure 1 The multiple cloning site MCS2 of the plasmid pPICZαABC3 was constructed, and a multi-copy system was constructed. Sequencing confirmed that the cDNA gene expression vector 4 of Trichoderma reesei endonuclease had been successfully constructed. This expression vector 4 was integrated into Pichia pastoris GS115. After screening on YPD plate, transfer to YPD liquid medium for culture. After the yeast is harvested, the enzyme protein is purified and screened to obtain a high-yield strain with high enzyme specific activity and high yield, that is, the recombinant Pichia reesei endonuclease EGII strain expressing cellulase.

Embodiment 3

[0037] As shown in the accompanying drawings of the description, the cDNA gene sequence of Trichoderma reesei endonuclease EGIII is from the 5' end to the 3' end, and the cDNA of Trichoderma reesei endonuclease containing the EcoRI-XbaI restriction site obtained by PCR amplification fragments, insert figure 1 The multiple cloning site MCS2 of the plasmid pPICZαABC3 was constructed, and a multi-copy system was constructed. Sequencing confirmed that the cDNA gene expression vector 4 of Trichoderma reesei endonuclease had been successfully constructed. This expression vector 4 was integrated into Pichia pastoris GS115. After screening on YPD plate, transfer to YPD liquid medium for culture. After the yeast is harvested, the enzyme protein is purified, and a high-yield strain with high enzyme specific activity and high yield is obtained by screening, that is, the recombinant Pichia pastoris 5-reesei endonuclease EGIII strain expressing cellulase.

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 genetic engineering and in particular to a recombined yeast strain expressing cellulase and applications thereof in producing the cellulose; a multi-copy cellulose gene is integrated into a secretory expression vector; an apoenzyme secretion expression plasmid is constructed; and pichia yeast recipient bacterium is led to obtain the recombined pichia yeast expressing the cellulose. The recombined pichia yeast expressing the cellulose is high in yield and high in activity and can be used for extracellularly secreting the apoenzyme.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a recombinant yeast strain expressing cellulase and an application thereof. Background technique [0002] At present, with the development of molecular genetics and molecular biology technology, domestic and foreign are trying to apply genetic engineering technology to transform and construct efficient cellulose degrading strains, because these strains have unique genetic characteristics and biological properties, along with the The scope of application of the enzyme is constantly expanding. Most of the cellulase production adopts Trichoderma reesei mutagenesis to screen high-yield strains, and it is refined and extracted through liquid deep-layer ventilation fermentation. The performance of the mutagenic strain is unstable, and after several passages, the high-yield activity is often lost, so it is necessary to repeatedly cultivate new strains, and the p...

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): C12N1/19C12N9/42C12R1/84
Inventor 陈战
Owner 陈战
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