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Method for efficiently preparing anhydroicaritin through enzymatic conversion

An icariin and enzyme conversion technology, which is applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of undisclosed enzyme conversion, high industrialization cost, low reaction concentration, etc. Intermediate product, low cost, simple operation effect

Pending Publication Date: 2021-03-26
金凤燮 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the article does not disclose the enzymatic conversion of Epimedin A, Epimedin B, and Epimedin C, and only studies the enzyme reaction substrate-icariin, which has low reaction concentration, large reaction vessel, and high industrialization cost. shortcoming
[0009] So far, there is no information on the use of organic solvent-resistant epimedium flavonoid glycosides (epimaridin A or epimedin B or epimedin C or icariin) in the presence of organic solvents. Reaction in the reaction system, related reports on the efficient preparation of icariin

Method used

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  • Method for efficiently preparing anhydroicaritin through enzymatic conversion
  • Method for efficiently preparing anhydroicaritin through enzymatic conversion

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Embodiment 1

[0025] The method for efficiently preparing icariin by enzymatic conversion of the present invention is prepared according to the following steps:

[0026] a. Aspergillus [ Aspergullus sp.39g bacteria, reference: Dongming Wang (Wang Dongming) et al., ProcessBiochemistry 47 (2012) 133–138 pages] the bacteria were inoculated in 1000 ml of liquid medium containing 5% corn steep liquor and 1% epimedium powder medium, cultured with stirring at 29°C for 7 days to obtain a fermentation liquid, namely the original enzyme liquid;

[0027] b. Centrifuge the original enzyme solution to remove impurities and collect the supernatant, add 3 times the volume of methanol solution to the supernatant, overnight, and centrifuge to collect the enzyme protein precipitate; dissolve the enzyme protein precipitate in 1 / 10 volume of the original enzyme solution, 0.04M, pH5.0 citric acid buffer solution, centrifuged to remove impurities, to obtain Aspergillus organic solvent-resistant epimedium fla...

Embodiment 2

[0034] a. Preparation of Epimedium flavonoidase resistant to organic solvents: Aspergillus niger[ Aspergullus niger g.848, reference: Yongkun Xiao (Xiao Yongkun), etc., J. Ginseng Res. 43 (2019) 186-195 pages] bacteria were inoculated in 400 grams (dry matter) of solid medium containing 10% licorice stem and leaf powder, Cultivate at 30°C (with occasional stirring) for 8 days; then use 2000 ml of normal saline to extract the enzyme in the solid medium, separate the supernatant, and filter to obtain the filtrate, which is the original enzyme solution;

[0035] b. Centrifuge the original enzyme solution to remove impurities and collect the supernatant, add 3 times the volume concentration of 95% ethanol to the supernatant, overnight, and centrifuge to collect the enzyme protein precipitate; dissolve the enzyme protein precipitate in the original enzyme solution volume 1 / 5. In the phosphate buffer solution of 0.04M and pH5.0, centrifuge to remove impurities to obtain the orga...

Embodiment 3

[0041] a. Preparation of Epimedium flavonoidase resistant to organic solvents: Candida was inoculated in 400 g (dry matter) of wheat bran solid medium containing 15% Sophorae pollen, cultured at 30°C (stirring occasionally) for 8 days; then Use 2000 ml of 0.02 mol and pH=5.0 acetate buffer to extract the enzyme in the solid medium, separate the supernatant, and filter to obtain the filtrate, which is the original enzyme solution;

[0042] b. Centrifuge the original enzyme solution to remove impurities and collect the supernatant, add 3 times the volume of methanol to the supernatant, overnight, and centrifuge to collect the enzyme protein precipitate; dissolve the enzyme protein precipitate in 1 / 8 of the original enzyme solution volume, 0.04M , in the acetic acid buffer solution of pH 5.0, and centrifuged to remove impurities to obtain the organic solvent-resistant epimedium flavonoid glycoside enzyme solution of Candida fungus;

[0043] c. Mix 100 ml of organic solvent-resis...

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Abstract

The invention discloses a method for efficiently preparing anhydroicaritin through an enzyme conversion. The method comprises the following steps: firstly, preparing organic solvent-resistant epimedium flavonoid glycosidase, and performing an enzyme reaction on the organic solvent-resistant epimedium flavonoid glycosidase and main epimedium flavonoid glycosides (epimedin A or epimedin B or epimedin C or icariin) in a reaction system containing an organic solvent. The method caneffectively reduce intermediate products and improve the conversion yield of the anhydroicaritin to 90% or above. Thereacted organic solvent-resistant epimedium flavonoid glycosidase can be recycled, a preparation cost of the anhydroicaritin is reduced, and the method has characteristics of simple operation, low cost, suitability for mass production and the like.

Description

technical field [0001] The invention relates to a method for preparing icariin, in particular to a method for efficiently preparing icariin by enzymatic conversion. Background technique [0002] Epimedium is a commonly used traditional Chinese medicine. Its stems and leaves are used as medicine, which has the effects of tonifying kidney and strengthening yang, strengthening muscles and bones, dispelling rheumatism, treating impotence and nocturnal emission, and rheumatic pain. More than 50 species of Epimedium have been found in the genus Epimedium, among which the medicinal Epimedium recorded in the 2015 edition of "Chinese Pharmacopoeia" is: Epimedium (Berberaceae Epimedium ( Epimedium brevicorum Maxim); Epimedium sagitta ( Epimedium sagittatum Maxim); Epimedium pilosa ( Epimedium pubescens Maxim); Korean Epimedium ( Epimedium koreanum Nakai) and Epimedium Wushan ( Epimedium wushanense T.S.Ying) and so on. The main active ingredient in Epimedium medicinal mat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/06C12R1/685C12R1/72
CPCC12P17/06
Inventor 金凤燮鱼红闪
Owner 金凤燮
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