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Galloyl glucosyltransferase CsUGT84A22 gene and encoded protein thereof and application

A technology of glucosyl-glucose and glucosyl-glucose, applied in the field of molecular biology, can solve the problems of cumbersome method, high cost, no market commodity product and the like

Inactive Publication Date: 2015-09-02
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The catalytic product of uridine diphosphate glucose-dependent galloylglucosyltransferase, 1-O-galloyl-β-D-glucoside (βG), is a substrate for the synthesis of ester-type catechins, but there is no commercial product so far product
In order to obtain 1-O-galloyl-β-D-glucoside (βG), it can be extracted from the fresh leaves of tea tree or obtained by chemical synthesis, but these two methods have the disadvantages of complicated methods, high cost and low yield. shortcoming

Method used

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  • Galloyl glucosyltransferase CsUGT84A22 gene and encoded protein thereof and application
  • Galloyl glucosyltransferase CsUGT84A22 gene and encoded protein thereof and application
  • Galloyl glucosyltransferase CsUGT84A22 gene and encoded protein thereof and application

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Embodiment 1

[0027] 1. Materials

[0028] 1. Tea tree variety: Nongkangzao (Camellia sinensis (L.) O.Kuntze.var.sinensis cultivar Nongkangzao), fresh tea leaves were collected, quickly frozen with liquid nitrogen, and stored in a -80°C refrigerator for later use;

[0029] 2. pMal-c2X vector: its plasmid map is as follows: figure 2 shown;

[0030] 3. Escherichia coli Novablue (DE3) expression host bacteria: purchased from Shanghai Beinuo Biotechnology Co., Ltd.;

[0031] 4. LB medium: Weigh 10g of NaCl, 5g of yeast extract, 10g of tryptone, add 950mL of ultrapure water and stir to dissolve, adjust the pH to 7.0 with 1mol / L NaOH, add water to 1000mL, and pressurize Steam sterilize for 15 minutes to obtain LB liquid medium, and LB solid medium can be obtained by adding 15 g of agar powder to LB liquid medium;

[0032] 5. Galactose solution with a mass concentration of 40%: weigh 40g of galactose, add ultrapure water to dissolve and stir evenly, set the volume to 100mL, and sterilize at 11...

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Abstract

The invention discloses a galloyl glucosyltransferase CsUGT84A22 gene with a nucleotide sequence shown as SEQ ID NO: 1. An encoded protein of the galloyl glucosyltransferase CsUGT84A22 gene is in an amino acid sequence shown as SEQ ID NO: 2. Compared with the prior art, the galloyl glucosyltransferase CsUGT84A22 gene has the advantages that the galloyl glucosyltransferase CsUGT84A22 gene for formation of uridine diphosphate glucose related to 1-O-galloyl-beta-D-glucoside are functionally cloned and verified for the first time, recombinant proteins, transgenic engineering bacteria and recombinant plasmids containing the CsUGT84A22 gene are provided, and mass synthesis of 1-O-galloyl-beta-D-glucoside is realized by means of bioengineering to lay the foundation for researches on biosynthesis regulation of ester catechins.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to a galloylglucosyltransferase CsUGT84A22 gene, its encoded protein and its application. Background technique [0002] Tea is one of the three major non-alcoholic beverages in the world. Catechin (flavan-3-ol), as the main health care and pharmacological components in tea, is the main tea polyphenols in tea, accounting for about 12–24% of the dry weight of fresh tea leaves, and is also the extract of black tea The typical bitter and astringent compounds in tea, especially epigallocatechin gallate (EGCG for short) have the highest content in tea soup and contribute the most to the astringency of tea, and give tea a bitter taste. [0003] Catechins are derivatives of 2-phenylbenzopyran and belong to flavan-3-ols in flavonoids. According to whether the 3-position on the C ring is connected with a gallo group, catechins can be divided into ester catechins (mainly including ep...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10C12N15/70C12N1/21C12P17/06
Inventor 刘亚军姚胜波崔利兰代新龙夏涛高丽萍
Owner ANHUI AGRICULTURAL UNIVERSITY
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