Recombinant escherichia coli capable of producing cinnamic alcohol and rosin, and construction method and applications thereof

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

Active Publication Date: 2018-06-26
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 biosynthe

Method used

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  • Recombinant escherichia coli capable of producing cinnamic alcohol and rosin, and construction method and applications thereof
  • Recombinant escherichia coli capable of producing cinnamic alcohol and rosin, and construction method and applications thereof
  • Recombinant escherichia coli capable of producing cinnamic alcohol and rosin, and construction method and applications thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0046] This example is used to illustrate the remodeling of the cinnamyl alcohol and / or rosin biosynthetic pathway 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, which can be various Escherichia coli commonly used in the art that can express the target gene, and the used 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 E. coli is preferably BL21(DE3).

[0048] The expression vectors in the present invention are preferably E. coli expression vector V1 and E. coli expression vector V2. E. coli expression vector V1 preferably contains genes PAL, CCR and 4CL, and E. coli expression vector V2 preferably contains genes UGT73B6 or UGT73C5 or oleD.

[0049] In the present invention, the order of the above-mentioned genes ...

Embodiment 2

[0066] This example is used to illustrate the fermentation culture 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 described above into the LB liquid medium (tryptone 10g / L, sodium chloride 10g / L, yeast extract 5g / L) containing antibiotics as selection markers, and cultivated at 37°C for 16 hours to obtain Escherichia coli seed liquid;

[0069] (2) inoculate the Escherichia coli seed liquid obtained in step (1) into 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 Induction culture for 16 hours.

[0070] It should be understood by those skilled in the art that during fermentati...

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Abstract

The invention discloses a recombinant escherichia coli capable of producing cinnamic alcohol and rosin, and a construction method and applications thereof. The construction method comprises followingsteps: phenylalnine ammonialyase PAL, p-hydroxycinnamic acid coenzyme A ligase 4CL, and cinnamamide coenzyme A reductase CCR are introduced into Escherichia coli so as to obtain the recombinant escherichia coli capable of producing cinnamic alcohol. In the construction method, Escherichia coli is taken as a model organism, the background is clear, growth speed is high, large scale fermentation culture is convenient to realize, and cost is low. The recombinant escherichia coli capable of producing cinnamic alcohol and rosin is obtained via construction of a cinnamic alcohol synthesis pathway, the cinnamic alcohol yield is 300mg/L; UDP glucosyltransferase is introduced into Escherichia coli further, so that organic combination of biological synthesis pathways of rosin and cinnamic alcohol isrealized, the highest rosin yield is 280mg/L. The construction method is capable of providing large scale industrialized production of cinnamic alcohol and/or rosin with foundation.

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 nephron, a construction method and an application. Background technique [0002] Rhodiola rosea is a traditional herbal medicine with physiological activities of anti-aging, anti-fatigue, enhancing immune function, protecting cardiovascular and cerebrovascular and anti-tumor. Rhodiola rosea is an emerging botanical medicine with environmental adaptability. Since the 1960s, scientists have carried out extensive research on its chemical active ingredients and pharmacological effects. The analysis found that its main active ingredients include salidroside, luoxai and its derivatives luoxevin and luoxelin. Luosai and its derivatives Luosaiwei and Luoceline have pharmacological activities such as antioxidant, improving physical endurance, relieving psychological stress and enhancing memory. Its agly...

Claims

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Application Information

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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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