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

A kind of construction method and application of engineering strain for producing phenol by fermentation of glycerol

A technology of engineering strains and glycerin, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of high production cost of phenol, large environmental pollution, and many by-products, so as to reduce production costs and improve economic efficiency. benefit effect

Active Publication Date: 2019-02-19
QUFU NORMAL UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problems of high production cost, large environmental pollution, and many by-products in the production of phenol by chemical methods in the above prior art, and the fact that there are few studies on the synthesis of phenol by biological methods, the present invention uses glycerin as a raw material to study a Construction method of an engineering strain producing phenol by fermentation of glycerol

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 kind of construction method and application of engineering strain for producing phenol by fermentation of glycerol
  • A kind of construction method and application of engineering strain for producing phenol by fermentation of glycerol
  • A kind of construction method and application of engineering strain for producing phenol by fermentation of glycerol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Construction of recombinant E. coli used for glycerol fermentation to produce phenol

[0058] 1. Construction of recombinant expression vector pTrc99a-tpl-p4m

[0059] 1. Selection and optimization of tpl gene

[0060] Select the complete CDS sequence of tyrosine phenol lyase (tpl), and obtain an optimized tyrosine phenol lyase gene sequence suitable for expression in E. coli after design and repeated verification, that is, the wild-type tyrosine phenol lyase gene The sequence is transformed into an E. coli preferred (high frequency use) codon optimized sequence without changing the amino acid sequence, thereby increasing the expression level of tyrosine phenol lyase in E. coli culture environment.

[0061] The amino acid sequence of the tyrosine phenol lyase is specifically shown in sequence 1 in the sequence table, and consists of 456 amino acids in total;

[0062] The coding gene sequence of the tyrosine phenol lyase is specifically shown in sequence 2 in the seque...

Embodiment 2

[0115] Example 2: Fermentation to produce phenol with glycerol as substrate

[0116] 1. The fermentation process of using glycerol to produce phenol

[0117] The E. coli engineered strain is preferably:

[0118] PH1: ATCC31884-pTrc99a-tpl-p4m;

[0119] PH2: ATCC31884-pTrc99a-tpl-p4m-trcglpX-tktA;

[0120] PH3: ATCC31884-pTrc99a-tpl-trcp4m-trcglpX-tktA;

[0121] The culture medium involved in the fermentation and production of phenol is preferably:

[0122] Solid plate medium (g / L): peptone 10, sodium chloride 10, yeast powder 5, agar 15, pH 7.0, add Amp 100μg / mL to the medium if necessary;

[0123] Fermentation seed medium (g / L): peptone 10, sodium chloride 10, yeast powder 5, pH 7.0, filling volume 10mL / 20mL, add Amp 100μg / mL to the medium if necessary;

[0124] Fermentation medium (g / L): glycerol 10, Na 2 HPO 4 ·12H 2 017.11, KH 2 PO 4 3. NaCl 0.5, NH 4 Cl 1, MgSO 4 ·7H 2 O0.492, CaCl 2 ·6H 2 O 0.022, filling volume 10mL / 20mL, add Amp 100μg / mL to the medium if necessary;

[0125] (1) Seed ...

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 technical field of engineered strain construction, in particular to a construction method of an engineered strain for producing phenol through glycerol fermentation. The second base sequence and the third base sequence in a sequence list are inserted in a pTrc99a carrier to obtain recombinant plasmid; the recombinant plasmid is electrically transformed into an ATC31884 strain, the engineered strain for producing the phenol through glycerol fermentation is obtained. The constructed engineered strain efficiently presents tyrosine phenol lyase and phenylalanine4-monooxygenase, and content of the obtained escherichia coli engineered strain for producing the phenol through glycerol is 21.30-57.66 mg / L, so that the phenol is produced through glycerol fermentation through the biological method, production cost of the phenol is reduced, and economic benefits are improved.

Description

Technical field [0001] The invention relates to the technical field of engineering strain construction, in particular to a method for constructing an engineering strain that uses glycerol to ferment to produce phenol, and also relates to the application of the obtained engineering strain. Background technique [0002] Phenol is an important basic organic chemical raw material, mainly used in the production of phenolic resin, bisphenol A and caprolactam, adipic acid, aniline, etc.; phenol derivatives such as halogenated phenols, nitrophenols, and alkylphenols can be used in medicine, The production of dyes, fragrances and various additives, so the phenol market has a good development prospect. [0003] Glycerin is also called glycerol, molecular formula C 3 H 5 (OH) 3 , Is a viscous liquid with sweet taste, can be miscible with water in any ratio, and has strong hygroscopicity. Glycerin is rich in resources and low in price. Because of its high reduction of C atoms, it has a higher...

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): C12N1/21C12N15/70C12P7/22C12R1/19
Inventor 车程川王斌王莹袁雪娇李敏
Owner QUFU NORMAL UNIV
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