Immobilization of Recombinant Strain Producing 2, 5-Furanedicarboxylic Acid and Its Application

A technology of genetically engineered bacteria and furandicarboxylic acid, applied in the direction of immobilized on/in organic carriers, based on microorganisms, bacteria, etc., can solve the problems of serious environmental pollution and high operating process requirements, and achieve broad application prospects, good reusability

Pending Publication Date: 2019-01-18
ANHUI RUISAI BIOCHEM TECH
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the raw materials of this method are relatively cheap and easy to obtain, the

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

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1,

[0034] Plasmids and strains:

[0035] Cloning vector pTE28aT7 - , E. Coli JM109 was purchased from a commercial company.

[0036] Restriction endonuclease, Taq enzyme and T4DNA ligase were all purchased from Dalian Bao Biological Engineering Company, and other chemical reagents were domestic analytical pure chemical reagents.

[0037] Glycerol dehydratase separation medium: 1000mL of distilled water, 6g of fructose, 5g of NaCl, 7g of yeast powder, 20g of glucose, 15g of agar powder, pH 7.0, 0.1MPa sterilization for 20min. Cool to 35℃~45℃, add Neil Red (Nile Red) 2mL / L (0.30mg Nile Red dissolved in 100mL dimethyl sulfoxide), pour it into a petri dish under aseptic conditions, and cool it for use.

[0038] Nutrient-rich medium: distilled water 1000mL, yeast powder 10g, agar powder 10g, fructose 3g, (NH 4 ) 2 SO 4 5g, adjust pH 7.0, 0.1MPa sterilization for 10min.

[0039] Product fermentation medium:

[0040] The preparation of phosphate buffer: distilled water 1000mL,...

Example Embodiment

[0056] Example 2,

[0057] The composition and culture conditions of each medium are the same as above, the preparation of recombinant E. coli is the same as the above, and the preparation method and conditions of the supramolecular self-assembly template are the same as the above.

[0058] The recombinant genetically engineered bacteria immobilized on the supramolecular self-assembly template after the fifth use was filtered from the fermentation broth, and the following fermentation experiment was performed.

[0059] Fermentation of recombinant genetically engineered bacteria to synthesize 2,5-furandicarboxylic acid:

[0060] Recombinant Escherichia coli preserved at -70℃ is activated on the LB medium plate, a single colony is picked with a sterilized toothpick and inoculated into 20ml LB medium, cultivated overnight at 31℃~37℃, and inoculated into the fermentation medium with 2% inoculum 1-2mmol / L IPTG was added to induce the expression of gldhA, and the fermentation was cultured f...

Example Embodiment

[0062] Example 3.

[0063] The composition and culture conditions of each medium are the same as above, the preparation of recombinant E. coli is the same as the above, and the preparation method and conditions of the supramolecular self-assembly template are the same as the above.

[0064] The recombinant genetically engineered bacteria fixed with the supramolecular self-assembly template after the 30th use was filtered from the fermentation broth, and the following fermentation experiment was performed.

[0065] Fermentation of recombinant genetically engineered bacteria to synthesize 2,5-furandicarboxylic acid:

[0066] Recombinant Escherichia coli preserved at -70℃ is activated on the LB medium plate, a single colony is picked with a sterilized toothpick and inoculated into 20ml LB medium, cultivated overnight at 31℃~37℃, and inoculated into the fermentation medium with 2% inoculum 1-2mmol / L IPTG was added to induce the expression of gldhA, and the fermentation was cultured for 48...

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 relates to the immobilization of recombinant strain producing 2, 5-furanedicarboxylic acid and the application thereof. The recombinant strain was constructed by integrating glycerol dehydratase gene gldabc of clostridium into gene dha beta of klebsiella terrestris to form recombinant dna gene gldha. The recombinant plasmid pte28at7 was amplified by pcr and inserted into escherichiacoli expression vector pte28at7 to construct 2, 5-recombinant furanedicarboxylic acid gene engineering strain pte28at7-gldha; supramolecular self-assembly template is a super-large specific surface area (no less than 1000m2/g) template with pore size of 20-35nm prepared by sodium silicate, cetyltrimethylammonium bromide, ammonium chloride and aluminum trichloride in aqueous phase and supramolecular self-assembly condition.

Description

[0001] Technical field: The invention belongs to microbial fermentation technology, and relates to the immobilization and application of recombinant genetically engineered bacteria producing 2,5-furandicarboxylic acid. Background technique [0002] 2,5-furandicarboxylic acid (FDCA) is an important intermediate in organic synthesis, which can be used to prepare various alkyl substituted or ester furan derivatives. Alkyl-substituted derivatives are widely used in the synthesis of chiral catalysts, molecular recognition receptors and polymer materials; ester derivatives are important fragrances, mainly used in food and cosmetic flavors. In addition, 2,5-furandicarboxylic acid can replace terephthalic acid to produce polyester-based plastic materials. In terms of pharmacology, diethyl 2,5-furandicarboxylate has an anesthetic effect similar to that of cocaine, and calcium salt of 2,5-furandicarboxylate has the function of inhibiting the growth of Bacillus megaterium. [0003] Di...

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): C12N1/21C12N11/02C12P17/04C12R1/19
CPCC12N9/88C12N11/02C12P17/04C12Y402/0103
Inventor 不公告发明人
Owner ANHUI RUISAI BIOCHEM TECH
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
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap