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Recombinant Escherichia coli for producing cinnamyl alcohol and rothium, construction method and application

A technology of recombinant Escherichia coli and a construction method, which is applied in the field of bioengineering, can solve the problems of no related reports on the heterologous biosynthesis of erythrocytes, achieve important economic value and social benefits, low cost, and rapid growth

Active Publication Date: 2021-07-20
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The heterologous biosynthesis of rosin has not been reported

Method used

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  • Recombinant Escherichia coli for producing cinnamyl alcohol and rothium, construction method and application
  • Recombinant Escherichia coli for producing cinnamyl alcohol and rothium, construction method and application
  • Recombinant Escherichia coli for producing cinnamyl alcohol and rothium, construction method and application

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Embodiment 1

[0046] This example is used to illustrate the remodeling of the biosynthetic pathway of cinnamyl alcohol and / or rospermia in Escherichia coli

[0047] In the present invention, there is no special requirement for the type of Escherichia coli used to construct the Escherichia coli expression strain, and it can be various Escherichia coli commonly used in the art that can express the target gene, and the used bacterial strain also has the alcohol dehydrogenase of Escherichia coli itself CAD, which reduces cinnamaldehyde to cinnamyl alcohol. In order to enable better expression of the target gene, the Escherichia coli is preferably BL21(DE3).

[0048] The expression vector among the present invention is preferably Escherichia coli expression vector V1 and Escherichia coli expression vector V2, Escherichia coli expression vector V1 preferably contains gene PAL, CCR and 4CL, Escherichia coli expression vector V2 preferably contains gene UGT73B6 or UGT73C5 or oleD.

[0049] In the ...

Embodiment 2

[0066] This example is used to illustrate the fermentation and cultivation of Escherichia coli recombinant strains BC, BR1, BR2 and BR3

[0067] In the present invention, the method for fermenting culture may comprise the following steps:

[0068] (1) Inoculate the Escherichia coli expression strain constructed as above into the LB liquid medium (tryptone 10g / L, sodium chloride 10g / L, yeast extract 5g / L) containing antibiotics as screening markers, and cultured at 37°C for 16 hours to obtain Escherichia coli seed liquid;

[0069] (2) Inoculate the Escherichia coli seed liquid obtained in step (1) to the M9 (Na 2 HPO 4 12H 2 O 15.12g / L, KH 2 PO 4 3.0g / L, NaCl 0.5g / L, MgSO 4 ·7H 2 O 0.5g / L, CaCl 2 0.011g / L, NH 4 Cl 1.0g / L, glucose 20g / L) liquid medium, and cultured at 37°C, when OD600 reached 0.6, add isopropyl-β-D-thiogalactopyranoside (IPTG), 16°C Under induction culture for 16 hours.

[0070] Those skilled in the art should understand that, during fermentation, the...

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Abstract

The invention discloses a recombinant Escherichia coli for producing cinnamyl alcohol and closone, its construction method and application. The construction method of the recombinant Escherichia coli for producing cinnamyl alcohol comprises the following steps: introducing phenylalanine ammonia-lyase PAL into Escherichia coli, para-hydroxy Cinnamoyl-CoA ligase 4CL and cinnamoyl-CoA reductase CCR were used to obtain a recombinant Escherichia coli producing cinnamyl alcohol. As a model organism, Escherichia coli has a clear background, rapid growth, easy large-scale fermentation and culture, and low cost; and the present invention obtains Escherichia coli that produces cinnamyl alcohol by constructing a cinnamyl alcohol synthesis pathway, and the output of cinnamyl alcohol reaches 300mg / L; UDP glucosyltransferase was further introduced to organically combine the biosynthetic pathways of rosopil and cinnamyl alcohol, and the maximum output of rospermia could reach 280mg / L. The invention lays the foundation for the large-scale industrial production of cinnamyl alcohol and / or rosopil.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a recombinant Escherichia coli for producing cinnamyl alcohol and closone, a construction method and application thereof. Background technique [0002] Rhodiola rosea is a traditional herbal medicine with anti-aging, anti-fatigue, immune function enhancement, cardiovascular and cerebrovascular protection and anti-tumor physiological activities. Rhodiola rosea is an emerging herbal medicine with environmental adaptability. Since the 1960s, scientists have conducted extensive research on its chemical active ingredients and pharmacological effects. The analysis found that its main active ingredients include salidroside, rosacein and its derivatives rosacein and rosacein. Luosai and its derivatives, Luosaiwei and Luosailin, have pharmacological activities such as anti-oxidation, improving physical endurance, relieving psychological stress and enhancing memory. It...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/70C12N1/21C12P7/22C12P19/44C12R1/19
CPCC12N9/0008C12N9/88C12N9/93C12N15/70C12N2800/101C12P7/22C12P19/44C12Y102/01044C12Y403/01005C12Y602/01012
Inventor 刘涛周威毕慧萍庄以彬殷华马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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