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Immobilized enzymatic-conversion large-flowered skullcap root and application thereof to acetaminophen damage repairing

A technology for immobilizing enzymes and baicalin, which is applied to immobilized enzymes, glycosylases, immobilized on/in organic carriers, etc., can solve the problems of easy inactivation of enzymes and high cost of enzymatic reactions, and achieve stable Good properties, improving enzyme content and stability, good stability and enzymatic activity

Inactive Publication Date: 2020-03-31
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The first object of the present invention is to provide a method for preparing immobilized enzymes to solve the technical problems of easy inactivation of enzymes and high cost of enzymatic reactions in the prior art

Method used

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  • Immobilized enzymatic-conversion large-flowered skullcap root and application thereof to acetaminophen damage repairing
  • Immobilized enzymatic-conversion large-flowered skullcap root and application thereof to acetaminophen damage repairing
  • Immobilized enzymatic-conversion large-flowered skullcap root and application thereof to acetaminophen damage repairing

Examples

Experimental program
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Effect test

Embodiment 1

[0056] The present embodiment provides a method for preparing an immobilized enzyme, comprising the following steps:

[0057] (1) Pretreatment of carrier material: Amberblite XAD1180N polystyrene macroporous adsorption resin was used as carrier material, Amberblite XAD1180N polystyrene macroporous adsorption resin was soaked in 95% ethanol aqueous solution for 10 hours, and stirred from time to time to make it Fully swollen, then the above-mentioned resin is continuously washed with distilled water until there is no obvious ethanol odor in the effluent; the resin and water are separated through a column to obtain a pretreated carrier material, which is stored in a 4°C freezer for later use;

[0058] (2) Weigh 80 mg of pectinase (Sangon Bioengineering (Shanghai) Co., Ltd., product batch number EC20BA0008) into a 10 mL centrifuge tube, add 6 mL of acetic acid-acetate buffer with pH=5 (0.02 mol / L) , dissolve, prepare the enzyme solution;

[0059] (3) Accurately weighing 0.5 g of...

Embodiment 2

[0063] The application of the immobilized enzyme of the present invention for biotransformation comprises the following steps:

[0064] 90 mg of the immobilized enzyme in Example 1 was mixed with the baicalin substrate (2.69 μM), and the transformation reaction was carried out on a shaking table at 50° C. for 4 h. The mass ratio of immobilized enzyme to baicalin substrate was 1:300.

[0065] The concentration of the baicalin substrate is not limited to this, for example, it can also be 1-10 μM or the like.

Embodiment 3

[0067] The application of the immobilized enzyme of the present invention for biotransformation comprises the following steps:

[0068] 70 mg of the immobilized enzyme in Example 1 was mixed with baicalin substrate (2.69 μM), and the transformation reaction was carried out on a shaking table at 40° C. for 8 h. The mass ratio of immobilized enzyme to baicalin substrate was 1:300.

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Abstract

The invention relates to the technical field of enzyme immobilization, in particular to an immobilized enzymatic-conversion large-flowered skullcap root and application thereof to acetaminophen damagerepairing. The preparation method comprises the following steps: mixing an enzyme solution with a pretreated carrier material and then carrying out absorption treatment; carrying out cross-linking treatment on the processed mixture and across-linking agent; and carrying out washing to obtain an immobilized enzyme. The carrier material is selected from macroporous resin. The enzyme contains pectinase. The immobilized enzyme provided by the invention has high enzyme content and stability and can be repeatedly recycled; and the production cost of enzymatic reaction can be lowered. The immobilized enzyme can be applied to biotransformation; the baicalin can be treated under mild reaction conditions to obtain a single product baicalein without any byproduct; and the product is easy to separate. The conversion product obtained by utilizing the immobilized enzyme has more remarkable protection and repair effects on the damage of acetaminophen to immune cells.

Description

technical field [0001] The invention relates to the technical field of enzyme immobilization, in particular to an immobilized enzyme to transform Scutellaria baicalensis and its application in repairing acetaminophen damage. Background technique [0002] After structural modification, natural active molecules can be converted into compound molecules with better drug activity and lower toxicity. The biotransformation pathway can modify the structure of natural drugs, which can improve the physicochemical properties of the drugs and further improve their efficacy. [0003] Flavonoids are important secondary metabolites in plants, show various physiological activities such as anti-aging, tumor, antioxidant, etc., and have important medicinal value. Flavonoids in plants mainly exist in the form of flavonoid glycosides. During plant growth, flavonoid glycosides are converted from flavonoid aglycones through the process of glycosylation. Depending on the number of sugar groups an...

Claims

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Application Information

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IPC IPC(8): C12N11/082C12P17/06A61K31/352A61P37/02
CPCA61K31/352A61P37/02C12N9/2408C12N11/08C12P17/06C12Y302/01015
Inventor 刘灿雷梦汪文漪谭黎马兰青薛飞燕杨明峰任艳艳张凌莉
Owner BEIJING UNIV OF AGRI
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