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Escherichia coli strain and method for biosynthesizing pyranocoumarin and furocoumarin by using escherichia coli strain

A kind of Escherichia coli, biosynthesis technology, applied in the field of synthetic biology, can solve the problems of complex extraction and purification process, high cost, low content, etc.

Active Publication Date: 2021-06-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The content of Indian quincetin, dihydro sagerenin, Japanese procuracin and lomatin in plants is very low, and the process of extracting these compounds from plants is complicated and expensive
In addition, the cultivation of plants is also affected by climate, agriculture, and geographical environment, which may also lead to inconsistent compounds isolated from different batches

Method used

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  • Escherichia coli strain and method for biosynthesizing pyranocoumarin and furocoumarin by using escherichia coli strain
  • Escherichia coli strain and method for biosynthesizing pyranocoumarin and furocoumarin by using escherichia coli strain
  • Escherichia coli strain and method for biosynthesizing pyranocoumarin and furocoumarin by using escherichia coli strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]Example 1 Plasmid construction of synthetic pathways for quincetin, dihydroeosanthin, Japanese procurerol and lomatin

[0035] Plasmid pCDF-Duet-ximD contains 1 gene in the benzopyran biosynthetic gene cluster: the ximD gene (monooxygenase) from Streptomyces xiamenensis 318; Two genes in the synthetic gene cluster: the ximD and ximE genes (cyclase) from Streptomyces xiamenensis 318. All genes were amplified by PCR, wherein the ximD gene was amplified using the Streptomycesxiamenensis 318 genome as a template, and the ximE gene was a gene (ximEsyn) obtained after codon optimization for Escherichia coli, and its sequence is shown in SEQ ID NO.1 .

[0036] The schematic diagram of the heterologous synthesis of Indian quincetin, dihydroeosanthin, Japanese procurerol and lomatin described in the present invention is as follows figure 1 shown.

[0037] Plasmids and primers used in the examples are shown in Table 1:

[0038] Table 1 Primer list

[0039]

[0040]

[0...

Embodiment 2

[0051] Example 2 Construction of Indian quincetin and dihydro-axerinin production strains

[0052] The plasmid pCDF-Duet-ximD was directly transformed into Escherichia coli BL21(DE3) by heat shock calcium transformation method to obtain strain XL01.

Embodiment 3

[0053] The construction of embodiment 3 japonica alcohol and lomatin production strain

[0054] The plasmid pCDF-Duet-ximD-ximE was transformed into Escherichia coli BL21(DE3) by heat shock calcium transformation method to obtain strain XM02.

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Abstract

The invention discloses an escherichia coli strain and a method for biologically synthesizing pyranocoumarin and furocoumarin by using the escherichia coli strain. The strain is escherichia coli BL21 (DE3) containing benzopyran biosynthesis related genes; and the benzopyran biosynthesis related gene comprises at least one of a ximD gene and a ximE gene. According to the invention, FAD-dependent monooxygenase XimD and cyclase XimE are introduced into Escherichia coli, and pyranocoumarin and furocoumarin are produced in Escherichia coli by inducible expression of the two genes and addition of a substrate at the same time. According to the invention, the biosynthesis of furocoumarin and pyranocoumarin in microorganisms is realized for the first time.

Description

technical field [0001] The invention belongs to the field of synthetic biology, and relates to an Escherichia coli strain and a method for biosynthesizing pyranocoumarin and furanocoumarin. Background technique [0002] The furanocoumarins (quincetin, dihydroapacetin) and the pyranocoumarins (precurol and lomatin) are natural products of plant origin. Experiments have shown that furanocoumarins generally have anti-insect larval activity, and quincetin has shown photorepair activity for DNA when ultraviolet radiation (Miyazawa, M., Tsukamoto, T., Anzai, J., and Ishikawa, Y .(2004) Insecticidal Effect of Phthalides and Furanocoumarins from Angelica acutiloba Against Drosophila Melanogaster,J.Agric.Food Chem.52,4401-4405; Beier,R.C.(1990)Natural Pesticides and Bioactive Components in Foods,Rev.Environx.Tooltam .113,47-137). The linear pyranocoumarin japonica alcohol is generally regarded as the active ingredient of the traditional Chinese medicine Angelica sinensis, which has...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12P17/18C12P17/04C12R1/19
CPCC12N15/70C12N9/0004C12N9/00C12P17/181C12P17/04C12N2800/22Y02A50/30
Inventor 徐岷涓徐俊步绪亮贺贝贝翁静怡
Owner SHANGHAI JIAO TONG UNIV
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