Emodin glycosyltransferase protein FtUGT73BE5 as well as encoding gene and application thereof

A technology encoding gene and emodin, applied in the field of emodin glycosyltransferase protein FtUGT73BE5 and its encoding gene and application, can solve the problems of high separation cost, high cost, and impure product

Active Publication Date: 2018-11-16
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To sum up, the modification of emodin glycosides involves large pollution and high cost by chemical methods, while the existing microbial or enzymatic methods have impure products and high separation costs
Although there have been reports on the biological or chemical synthesis of emodin glycosides, these methods are either environmentally unfriendly or the diversity of target products

Method used

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  • Emodin glycosyltransferase protein FtUGT73BE5 as well as encoding gene and application thereof
  • Emodin glycosyltransferase protein FtUGT73BE5 as well as encoding gene and application thereof
  • Emodin glycosyltransferase protein FtUGT73BE5 as well as encoding gene and application thereof

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Embodiment 1

[0035] Embodiment 1, clone the coding gene of emodin glycosyltransferase

[0036] 1. Methods and steps of gene cloning

[0037] The collection of tartary buckwheat (Jinqiao No. 2, provided by Professor Chen Qingfu of Guizhou Normal University, the non-patent literature that recorded Jinqiao No. 2 is: Li Ruiyuan, Shi Taoxiong, Chen Qijiao, Pan Fan, Chen Qingfu (2017) 35 tartary buckwheat in China EST-SSR fingerprint construction and genetic diversity analysis of approved varieties, Acta Plant Science, 35, 267-275.) Leaves and seeds, RNA was extracted, reverse transcribed into cDNA, primer sequences (as shown in Table 1) were designed, and leaves and seeds were extracted The mixed cDNA was used as a template and cloned into the FtUGTs gene fragment using KOD high-fidelity enzyme (the total volume of the KOD high-fidelity enzyme PCR system is 50 μL: 5 μL 10XBuffer, 3 μL MgSO 4, 5 μL dNTP (2 mM), 5 μL primer (10 mM), 1 μL template and 8 μL water, the program is shown in Table 2)....

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Abstract

The invention discloses emodin glycosyltransferase protein FtUGT73BE5 as well as an encoding gene and an application thereof. The protein is as shown in the following a) or b): a) protein consisting of the amino acid sequence shown in the sequence 2 in a sequence table; b) protein which consists of the amino acid sequence shown in sequence 2 in the sequence table by substitution and / or deletion and / or addition of one or more amino acid residues, has emodin 6-O-glucoside glycosyltransferase activity and is derived from a). The emodin glycosyltransferase sequence is mainly obtained on the basisof Fagopyrum tataricum genome, and prokaryotic expression of recombinant protein confirms that FtUGT73BE5 can catalyze generation of emodin 6-O-glucoside from emodin. Meanwhile, the gene can be overexpressed in hairy roots through agrobacterium mediation by means of FtUGT73BE5, and finally, emodin 6-O-glucoside is obtained.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the emodin glycosyltransferase protein FtUGT73BE5 and its coding gene and application. Background technique [0002] The chemical name of Emodin is 1,3,8-trihydroxy-6-methylanthraquinone (1,3,8-Trihydroxy-6-methylanthraquinone). Molecular formula is C 15 h 10 o 5 , almost insoluble in ethanol and alkali solution. Emodin is a relatively specific compound in Polygonaceae plants. It exists in rhubarb, Polygonum multiflorum and Tartary buckwheat. It is rich in resources and has a wide range of sources, and its extraction method and purification process are relatively simple. Pharmacological experimental studies have shown that emodin has antibacterial, purging, diuretic, antispasmodic, anti-inflammatory, cough-relieving, and anti-tumor effects. At the same time, it is also beneficial to the treatment of type 2 diabetes clinically, and is widely used in health care products such as we...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/82A01H5/00A01H6/00C12P19/56
CPCC12N9/1051C12N15/8243C12P19/56
Inventor 陈士林尹青岗孙伟韩小燕师玉华
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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