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

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 operation process requires high requirements and the environmental pollution is serious.

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

Embodiment 1

[0034] Plasma particles and strains:

[0035] Cloning vector pTE28aT7 - , Escherichia coli E.Coli JM109 were purchased from commercial companies.

[0036] Restriction enzymes, Taq enzymes, and T4 DNA ligase were purchased from Dalian Bao Biological Engineering Company, and other chemical reagents were domestic analytically pure chemical reagents.

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

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

[0039] Product fermentation medium:

[0040] Preparation of phosphate buffer: dis...

Embodiment 2

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

[0058] The recombinant genetically engineered bacteria immobilized with the supramolecular self-assembly template after the fifth use were filtered out from the fermentation broth, and the following fermentation experiments were carried out.

[0059] Fermentative synthesis of 2,5-furandicarboxylic acid by recombinant genetically engineered bacteria:

[0060] The recombinant Escherichia coli stored at -70°C was activated on the LB medium plate, picked a single colony with a sterilized toothpick and inoculated in 20ml LB medium, cultured overnight at 31°C-37°C, and inoculated in the fermentation medium with 2% inoculum , adding 1-2 mmol / L IPTG to induce the expression of gldhA, and fermented for 48 hours.

[0061] Separa...

Embodiment 3

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

[0064]The recombinant genetically engineered bacteria immobilized with the supramolecular self-assembly template after the 30th use were filtered out from the fermentation broth, and the following fermentation experiments were carried out.

[0065] Fermentative synthesis of 2,5-furandicarboxylic acid by recombinant genetically engineered bacteria:

[0066] The recombinant Escherichia coli stored at -70°C was activated on the LB medium plate, picked a single colony with a sterilized toothpick and inoculated in 20ml LB medium, cultured overnight at 31°C-37°C, and inoculated in the fermentation medium with an inoculum of 2%. , adding 1-2 mmol / L IPTG to induce the expression of gldhA, and fermented for 48 hours.

[0067] S...

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 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