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Esculin and fraxin glycosyltransferase protein as well as coding gene and application thereof

A technology of aescin glycosyl and coding gene, which is applied in the field of escin and aquinoside glycosyltransferase protein and its coding gene and application, and can solve the problems of biosynthesis of coumarin, etc.

Active Publication Date: 2022-02-11
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

Unknown factors make biosynthesis of coumarin difficult

Method used

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  • Esculin and fraxin glycosyltransferase protein as well as coding gene and application thereof
  • Esculin and fraxin glycosyltransferase protein as well as coding gene and application thereof
  • Esculin and fraxin glycosyltransferase protein as well as coding gene and application thereof

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Embodiment 1

[0033] Embodiment 1, clone the coding gene of coumarin glycosyltransferase

[0034] 1. Methods and steps of gene cloning

[0035] Collect the horse chestnut leaves, flowers and seeds in the mature period, and analyze the coumarin content therein ( figure 2 ), found that the flowers are rich in aescin, aescin, aescin and aescin (Wei Yiding, Xiong Chao, Zhang Tianyuan, et al. Research on the synthesis pathway of triterpenoid saponins in Aesculus aesculus based on transcriptome data[J]. Chinese Traditional Medicine Journal, 2019(6):1135-1144.).

[0036] Therefore, using the above plant materials, RNAseq was performed with the aid of the Illumina platform. Tissue expression profile RNAseq data in horse chestnut showed that 19 highly expressed AcUGTs sequences were obtained. Based on the transcriptome data, combined with the AcUGTs conserved domain PSPG box, 19 AcUGTs sequences of 300aa-500aa were obtained, of which 1 AcUGTs was expressed in flowers very high.

[0037] Design ...

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Abstract

The invention discloses aesculin and fraxin glycosyl transferase protein as well as a coding gene and application thereof. The esculetin and fraxin glycosyl transferase protein provided by the invention is a protein as shown in a) or b): a) a protein consisting of an amino acid sequence as shown in a sequence 2 in a sequence table; b) a protein which is obtained by substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence as shown in the sequence 2 in the sequence table, has the activity of aesculin and / or fraxin glycosyltransferase and is derived from a). Based on related results of high-throughput sequencing of aesculus chinensis, a key enzyme UGTs in the last step of synthesis of fraxin and aesculin is found and identified by using a reverse genetics method, the terminal blank of a coumarin biosynthesis pathway is filled, and glycosyl transferase protein and a coding sequence thereof are provided for further biosynthesis of fraxin and aesculin.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to escin and arugeridin glycosyltransferase proteins, coding genes and applications thereof. Background technique [0002] Aesculus (Aesculuschinensis) is a plant of the genus Aesculus in the family Aesculus, and there are about 30 species of this genus in the world, including about 11 species in my country. Coumarin is an important small-molecule active ingredient in horse chestnut. It has anti-inflammatory, analgesic, antibacterial and antiviral pharmacological effects, can regulate plant growth, and is used as a phytoalexin. It has high bioavailability in the human body. The coumarins that have been isolated from horse chestnut include Esculetin, Esculin, Fraxin and Fraxetin ( figure 1 )Wait. These four coumarin components are not available as medicines, so the research on the biosynthetic pathway of this coumarin will have important medical significance and economic value. Pharma...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54C12P19/46
CPCC12N9/1048C12P19/46Y02A50/30
Inventor 陈士林尹青岗孙伟魏一丁
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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