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Method for producing continuously 4-hydroxycinnamyl alcohol using multi-strain immobilized cell composition

A technology of immobilized cells and hydroxycinnamyl alcohol, which is applied in the field of enzyme engineering, can solve the problems of immobilization of a single bacterial strain limited by research, and achieve the effects of reducing the culture process of free cells, reducing the generation of by-products, and simplifying the production process

Inactive Publication Date: 2016-09-07
BEIJING FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are many studies on the existing immobilization technology, but most studies are limited to the immobilization of a single strain

Method used

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  • Method for producing continuously 4-hydroxycinnamyl alcohol using multi-strain immobilized cell composition
  • Method for producing continuously 4-hydroxycinnamyl alcohol using multi-strain immobilized cell composition
  • Method for producing continuously 4-hydroxycinnamyl alcohol using multi-strain immobilized cell composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Strain culture

[0039] The strains used are A strain and B strain;

[0040] A strain is a recombinant strain E. coli M15-4Cl-CCR, after induction, can highly express the fusion bifunctional enzyme 4Cl-CCR, has the catalytic activity of 4Cl and CCR enzymes, and can convert 4-hydroxycinnamic acid into 4-hydroxycinnamaldehyde;

[0041] B strain is a recombinant strain E. coli M15-CAD; can highly express CAD after induction, and can convert 4-hydroxycinnamaldehyde into 4-hydroxycinnamyl alcohol;

[0042] LB solid medium, in g / L, tryptone 10, yeast extract 5, NaCl10, pH7.5, ampicillin 0.1, kanamycin 0.025; LB liquid medium, in g / L, tryptone 10 , yeast extract 5, NaCl10, agar powder 15, pH7.5, ampicillin 0.1, kanamycin 0.025;

[0043] 1.1, the culture of strain A, the recombinant strain E. coli M15-4Cl-CCR was inoculated in LB solid medium and activated; a single colony of the recombinant strain was picked and inoculated in 30 mL of LB liquid medium containing 100 μ...

Embodiment 2

[0059] 1. Strain culture

[0060] The strains used are A strain and B strain;

[0061] A strain is a recombinant strain E. coli M15-4Cl-CCR, after induction, can highly express the fusion bifunctional enzyme 4Cl-CCR, has the catalytic activity of 4Cl and CCR enzymes, and can convert 4-hydroxycinnamic acid into 4-hydroxycinnamaldehyde;

[0062] B strain is a recombinant strain E. coli M15-CAD; can highly express CAD after induction, and can convert 4-hydroxycinnamaldehyde into 4-hydroxycinnamyl alcohol;

[0063] LB solid medium, in g / L, tryptone 10, yeast extract 5, NaCl10, pH7.5, ampicillin 0.1, kanamycin 0.025; LB liquid medium, in g / L, tryptone 10 , yeast extract 5, NaCl10, agar powder 15, pH7.5, ampicillin 0.1, kanamycin 0.025;

[0064] 1.1, the culture of strain A, the recombinant strain E. coli M15-4Cl-CCR was inoculated in LB solid medium and activated; a single colony of the recombinant strain was picked and inoculated in 30 mL of LB liquid medium containing 100...

Embodiment 3

[0080] 1. Strain culture

[0081] The strains used are A strain and B strain;

[0082] A strain is a recombinant strain E. coli M15-4Cl-CCR, after induction, can highly express the fusion bifunctional enzyme 4Cl-CCR, has the catalytic activity of 4Cl and CCR enzymes, and can convert 4-hydroxycinnamic acid into 4-hydroxycinnamaldehyde;

[0083] B strain is a recombinant strain E. coli M15-CAD; can highly express CAD after induction, and can convert 4-hydroxycinnamaldehyde into 4-hydroxycinnamyl alcohol;

[0084] LB solid medium, in g / L, tryptone 10, yeast extract 5, NaCl10, pH7.5, ampicillin 0.1, kanamycin 0.025; LB liquid medium, in g / L, tryptone 10 , yeast extract 5, NaCl10, agar powder 15, pH7.5, ampicillin 0.1, kanamycin 0.025;

[0085] 1.1, the culture of strain A, the recombinant strain E. coli M15-4Cl-CCR was inoculated in LB solid medium and activated; a single colony of the recombinant strain was picked and inoculated in 30 mL of LB liquid medium containing 100...

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Abstract

The invention relates to a method for producing continuously 4-hydroxycinnamyl alcohol using a multi-strain immobilized cell composition. Strain A used herein is recombinant strain E. coli M15-4Cl-CCR, and strain B is recombinant strain E. Coli M15-CAD; the method comprises the specific steps: 1, culturing strains; 2, immobilizing the strains to obtain respectively strain A immobilized cell particles and strain B immobilized cell particles; 3, producing 4-hydroxycinnamyl alcohol with the immobilized cells. The 4-hydroxycinnamyl alcohol is produced using modern bioengineering technology, and the application of the method for producing continuously with the multi-strain immobilized cell composition in the synthesis of plant-origin metabolite has important practical significance; high-performance liquid chromatography tandem mass spectrometry detection is established in the meantime, and more effective analytical means is provided for the aromatic polymers.

Description

technical field [0001] The invention belongs to the technical field of enzyme engineering, and in particular relates to a method for producing 4-hydroxycinnamyl alcohol by adopting cell immobilization technology. Background technique [0002] 4-Hydroxycinnamyl alcohol is a class of plant natural metabolites, including coumaryl alcohol, cafestol, coniferyl alcohol and sinapyl alcohol. They are intermediates in the biosynthesis of various plant secondary metabolites, such as vanillin and lignans. Among them, coumaryl alcohol, coniferyl alcohol and sinapyl alcohol are the three basic building blocks of lignin, which play an important role in water transport, mechanical support and plant pathogen defense. In addition, such alcohols also have important medicinal value. For example, coniferyl alcohol is also an important intermediate in the synthesis of silibinin, which can be used to treat liver diseases. The study of 4-hydroxycinnamyl alcohols is necessary to explore its rela...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/22C12N11/10G01N30/02C12R1/19
CPCC12P7/22C12N11/10G01N30/02G01N2030/027
Inventor 盖颖刘淑欣蒋湘宁
Owner BEIJING FORESTRY UNIVERSITY
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