Methyltransferase gene of glycosides

A technology of methyltransferase and compound, which is applied in the direction of transferase, plant gene improvement, microorganism-based methods, etc., can solve the problem of activity reduction and achieve the effect of enhancing stability and enhancing stability

Inactive Publication Date: 2018-02-09
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Various natural and unnatural glycosylated substances are easily affected by various factors from organisms and the outside world, resulting in self-degradation and reduced activity

Method used

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  • Methyltransferase gene of glycosides
  • Methyltransferase gene of glycosides
  • Methyltransferase gene of glycosides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Synthesis of O-methyltransferase gene BbFkbM, and construction of yeast expression vector to obtain yeast transformants

[0087] Sequencing the genome of a strain of Beauveria bassiana obtained, and obtained the genome sequence of the strain. During the process of analyzing glycosyltransferases through relevant bioinformatics software, an O-methyltransferase gene was found through comparison. We named it BbFkbM. The nucleotide sequence of BbFkbM was predicted and analyzed by various software to remove intron splicing to obtain the coding region sequence of BbFkbM. Extract the RNA of Beauveria bassiana and obtain cDNA by reverse transcription, use the designed primers to amplify the coding gene of BbFkbM, recover the target fragment and send it to the sequencing company for sequencing and comparison, and obtain the glycosyltransferase shown in SEQ ID NO.1 Gene BbFkbM.

[0088] Restriction sites are introduced during synthesis. An NdeI restriction site was ad...

Embodiment 2

[0092] Example 2 Using Glucose O-methyltransferase Recombinant Vector PXW06F-BbFkbM to Realize the Modification of Desmethyl-Lasiodiplodin Glycosylated Derivatives of Polyketides

[0093] 1. Purpose of the experiment

[0094] The metabolites of modified O-methyltransferase and glucosyltransferase were separated by HPLC and their molecular structures were analyzed.

[0095] 2. Experimental method:

[0096] 1) Fermentation culture

[0097] Adopt two-step fermentation technology, first inoculate an appropriate amount of yeast transformant cells into the corresponding 25ml - Leu / - In Trp liquid deficient medium, culture at 30°C and 200r min-1 for about 16 hours, then add 25ml of YPD low-sugar medium, and add 5mg of pure Desmethyl-Lasiodiplodin at the same time to continue culturing for 48 hours; extract the fermentation product with ethyl acetate, ethyl acetate The ratio of ester to fermentation broth is 1:1, that is, 50ml of ethyl acetate is used to extract the fermentation ...

Embodiment 3

[0111] Example 3 Application of Glucose O-methyltransferase Recombinant Vector PXW06F-BbFkbM to Realize the Modification of Flavonoids

[0112] 1. Purpose of the experiment

[0113] The metabolites of modified O-methyltransferase and glucosyltransferase were separated by HPLC and their molecular structures were analyzed.

[0114] 2. Experimental method:

[0115] 1) Fermentation culture

[0116] Adopt two-step fermentation technology, first inoculate an appropriate amount of yeast transformant cells into the corresponding 25ml - Leu / - In Trp liquid deficient medium, culture at 30°C and 200r min-1 for about 16 hours, then add 25ml of YPD low-sugar medium, and at the same time add 5mg of pure product to continue culturing for 48 hours. A total of 20 flavonoid samples were used; extracted with ethyl acetate For the fermentation product, the ratio of ethyl acetate to fermentation broth is 1:1, that is, the fermentation product is extracted with 50ml of ethyl acetate; the dry e...

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Abstract

The invention relates to a methyltransferase gene of glycosides. For the methyltransferase gene of glycosides, methylated modification can be carried out on four hydroxyl radicals of glycosyl in glycoside compounds in vitro, meanwhile, a synergistic effect can be achieved with glucosyltransferases, biosynthesis and bioconversion of different active substances including polyketides, flavonoids andanthraquinone are realized through combination, thus novel active compounds different from natural product structures are obtained, the oriented bioconversion of the compounds is realized, and the stability of the compounds is enhanced.

Description

Technical field: [0001] The invention relates to a gene capable of methylating the 4-hydroxyl group of glucose in glycosylated substances, and the glycosylated substances specifically include polyketones, flavonoids, and -glycosylated derivatives. Background technique: [0002] Various natural and unnatural glycosylated substances are easily affected by various factors from the organism and the outside world, resulting in self-degradation and reduced activity. If the 4-hydroxyl of the sugar group in these substances is alkylated, it is beneficial to enhance their stability. [0003] Fungal biotransformation is the use of specific enzymes in cells to modify and transform the structure of exogenous compounds to obtain more valuable metabolic reactions. Invention content: [0004] The purpose of the present invention is to obtain the O-methyltransferase gene that can participate in the methylation of the 4-position hydroxy group of the sugar group in the glycosyl compound, r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C12N15/81C12N1/19C12P19/62C12R1/865
CPCC12N9/1007C12N15/81C12P19/62
Inventor 张礼文徐玉泉谢李楠王辰王晓婧
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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