Glycosidase, coding gene of glycosidase and application of glycosidase and coding gene

A glycosidase and gene technology, applied in the field of genetic engineering, can solve problems such as low conversion rate, low enzyme activity, and unsatisfactory

Active Publication Date: 2021-04-02
KUNMING NOVOGINSENG BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing enzymes used for the biotransformation of ginsenosides are relatively low in activity, and most of them are highly specific, that is, they can only act on a certain glycoside structure specifically, but cannot simultaneously act on more than one glycoside structure. requirements, resulting in lower conversion rates

Method used

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  • Glycosidase, coding gene of glycosidase and application of glycosidase and coding gene
  • Glycosidase, coding gene of glycosidase and application of glycosidase and coding gene
  • Glycosidase, coding gene of glycosidase and application of glycosidase and coding gene

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preparation example Construction

[0055] In the sixth aspect, the present invention provides a preparation method of medicinal koji, which is characterized in that the preparation method comprises: contacting koji-making raw materials with glycosidase to obtain medicinal koji containing ginsenoside compound K;

[0056] Wherein, the koji-making raw material is a raw material containing ginsenoside and / or gypenoside, and the glycosidase is one or more glycosidases described in the first aspect or provided by the composition described in the fourth aspect.

[0057] The ginsenosides and / or gypenosides mentioned in the present invention refer to various ginsenosides and / or gypenosides common in the art contained in raw materials such as ginseng root, ginseng stem and leaf flower and notoginseng stem and leaf flower, such as ginsenoside Rb 1 , Ginsenoside Rb 2 , Ginsenoside Rb 3 , Ginsenoside Rc, Ginsenoside Rd, Ginsenoside F2, Ginsenoside Rg 3 , Ginsenoside Rh 2 , Ginsenoside C-O, Ginsenoside C-Mx 1 , Ginsenosi...

Embodiment 1

[0065] This example is used to illustrate the preparation of the glycosidase provided by the present invention.

[0066] (1) Acquisition of genes

[0067] According to the nucleotide sequence of the glycosidase (the corresponding number and the encoded amino acid sequence are as follows), the corresponding genes were obtained by artificial chemical synthesis (entrusted to Kunming Shuoqing Biotechnology Co., Ltd.).

[0068] (I) β-glucosidase

[0069] Nucleotide sequence (5'-3') of β-glucosidase:

[0070]

[0071] Amino acid sequence of β-glucosidase:

[0072]

[0073]

[0074] (II) Arabinosidase pyranosidase

[0075] Nucleotide sequence (5'-3') of arabinosidase:

[0076]

[0077] Amino acid sequence of arabinosidase:

[0078]

[0079] (III) Furanotype arabinosidase

[0080] Nucleotide sequence (5'-3') of furanotype arabinosidase:

[0081]

[0082] Amino acid sequence of arabinosidase furanosidase:

[0083]

[0084] (IV) Rhamnosidase

[0085] Nucleo...

Embodiment 2

[0106] This example is used to illustrate the application of the enzyme provided by the present invention in the preparation of koji containing ginsenoside CK.

[0107] Preparation of koji raw materials: according to the feeding amount: 12.5% ​​by weight of bran, 7.5% by weight of rice husk, 80% by weight of dry pulverized matter (400 orders of particle size) of Panax notoginseng, add water in the bran mixing machine and mix the raw materials evenly, and set aside;

[0108] Bran steaming: The material is evenly fed into the bran steaming machine, and steam is fed while feeding. After loading the material, turn on the atmosphere to steam the bran, steam for 60 minutes after turning off the steam, and then stew the bran for 20 minutes after turning off the steam.

[0109] Cooling and inoculation: After steaming, let the machine cool down to 50°C immediately, add the compound enzyme preparation (the dosage of the compound enzyme preparation is 4U per gram of the koji-making raw ma...

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Abstract

The invention relates to the field of gene engineering, and discloses glycosidase, a coding gene of the glycosidase and application of the glycosidase and the coding gene. Specifically, the inventionprovides the glycosidase, the coding gene of the glycosidase, a recombinant vector, an expression cassette, a transgenic cell line or recombinant bacteria containing the gene, and application of the glycosidase in degradation of glycoside compounds or preparation of medicated koji. When the glycosidase provided by the invention is used for degrading ginsenoside and/or gypenoside, ginsenoside CK can be obtained, and the conversion rate is relatively high.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to glycosidases and their coding genes and their application. Background technique [0002] Ginsenoside Compound K (ginsenoside Compound K, ginsenoside CK, structural formula shown in the following formula) is the main metabolite of diol-type ginsenosides in the human intestinal tract, and belongs to rare ginsenosides. Studies have found that ginsenoside CK has a good effect on inhibiting the growth and metastasis of tumor cells both in vivo and in vitro. Therefore, how to obtain a large amount of ginsenoside CK is the focus of current pharmaceutical research. [0003] [0004] Currently, ginsenoside CK is mainly obtained through biotransformation or biosynthesis. Among them, the biotransformation method is a process in which organisms (cells, organelles) or enzymes are used as catalysts to achieve chemical transformation, and it is a chemical reaction in which biological sys...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N9/42C12N15/56C12P33/20C12G3/026
CPCC12N9/2402C12N9/2445C12N9/2485C12P33/20C12G3/026C12Y302/01021C12Y302/0104C12Y302/01043
Inventor 包崇卯许云辉毕春艳刘兴周国婷
Owner KUNMING NOVOGINSENG BIOENG
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