Process of catalytically synthesizing salidroside from beta-glucosaccharase crosslinking aggregate

A technology of glucosidase and salidroside, which is applied in the field of enzyme-catalyzed synthesis of salidroside, can solve the problems of large amount of ionic liquid added, high cost of salidroside, complicated reaction steps and the like, and achieves simple recovery operation, Simple production and optimized reaction conditions

Active Publication Date: 2014-03-19
福建东亮生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a process for catalyzing the synthesis of salidroside by β-glucosidase cross-linked aggregates (abbreviation: β-glucosidase CLEAs) that can greatly increase the concentration of the product salidroside. The reaction process of the present invention does not require Expensive glycoside donors and catalysts avoid the cumbersome reaction steps of chemical synthesis. In addition, the target product salidroside can be easily extracted from the reaction solution after the reaction is completed, and the unreacted substrate p-hydroxyphenylethanol can be recycled again , reduce the production cost, and at the same time solve the problem that the cost of producing salidroside is high due to the high price of the ionic liquid, and the addition of the ionic liquid is still very large in the process. Therefore, the enzyme-catalyzed synthesis of salidroside is Tianglycoside mass production has great economic and practical significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] β-glucosidase cross-linked aggregates (abbreviated as β-glucosidase CLEAs) catalyze the process of synthesizing salidroside, including the preparation of β-glucosidase CLEAs and the synthesis of salidroside;

[0021] The preparation stage of β-glucosidase CLEAs includes the following steps: (1) Add 1.7ml 1,4-dioxane to 1ml β-glucosidase solution containing 2.5mg / ml β-glucosidase (1 , the volume ratio of 4-dioxane to β-glucosidase solution is 1.7:1 (v:v)), shake; (2) add glutaraldehyde to the final concentration of 38mM, react for 12min, carry out (3) Add sodium borohydride (NaBH 4 ), to NaBH 4 The final concentration of 25mM was reduced, after shaking, centrifuged at 300rpm for 8min, and the precipitate was collected, which was β-glucosidase CLEAs; the reaction temperature during the whole reaction process was controlled at 6°C, and the pH was 5.2; the precipitate was collected, and the β-glucosidase was measured. The recovery rate of the enzyme activity of glucosidas...

Embodiment 2

[0028] β-glucosidase CLEAs catalyzes the process of synthesizing salidroside, including the preparation of β-glucosidase CLEAs and the synthesis of salidroside;

[0029] The preparation stage of β-glucosidase CLEAs includes the following steps: (1) Add 2.2ml 1,4-dioxane to 1ml β-glucosidase solution containing 1.5mg / ml β-glucosidase (1 , the volume ratio of 4-dioxane to β-glucosidase solution is 2.2:1), shake; (2) add glutaraldehyde to a final concentration of 40mM, react for 8min, and carry out cross-linking; (3) Sodium borohydride (NaBH 4 ), to NaBH 4 The final concentration of 20 mM was reduced, after shaking, centrifuged at 500rpm for 5min, and the precipitate was collected, which was β-glucosidase CLEAs; the reaction temperature during the whole reaction process was controlled at 4°C, and the pH was 5.5; the precipitate was collected to measure β-glucosidase CLEAs. -The recovery rate of the enzyme activity of glucosidase CLEAs was 81.65%.

[0030] The synthesis stage o...

Embodiment 3

[0033] β-glucosidase CLEAs catalyzes the process of synthesizing salidroside, including the preparation of β-glucosidase CLEAs and the synthesis of salidroside;

[0034] The preparation stage of β-glucosidase CLEAs includes the following steps: (1) Add 2.0ml 1,4-dioxane to 1ml β-glucosidase solution containing 1.5mg / ml β-glucosidase (1 , the volume ratio of 4-dioxane to β-glucosidase solution is 2:1), shake; (2) add glutaraldehyde to a final concentration of 43mM, react for 8min, and carry out cross-linking; (3) Sodium borohydride (NaBH 4 ), to NaBH 4 The final concentration of 23 mM was reduced, after shaking, centrifuged at 400rpm for 5min, and the precipitate was collected, which was β-glucosidase CLEAs; the reaction temperature during the whole reaction process was controlled at 5°C, and the pH was 5.0; the precipitate was collected to measure β-glucosidase CLEAs. -The recovery rate of the enzyme activity of glucosidase CLEAs was 80.59%.

[0035] The synthesis stage of ...

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Abstract

The invention provides a process of catalytically synthesizing salidroside from beta-glucosaccharase crosslinking aggregate (beta-glucosaccharase CLEAs); in the synthesis stage of the salidroside, a reaction system is a 1-butyl-3-methylimidazole hexafluorophosphate/water-mimicking solvent/1,4-dioxane/buffer solution, the water-mimicking solvent is ethyl acetate, the unit enzyme activity of the beta-glucosaccharase CLEAs is 1.3-1.6U/ml, the ratio of the volume summation of 1-butyl-3-methylimidazole hexafluorophosphate and ethyl acetate to the volume of the 1,4-dioxane is 2: 3-3: 2, the volume percent in the buffer solution is 12-18%, the reaction temperature is 48-52 DEG C, and the reaction time is 5-9 days. The concentration of the product salidroside can be greatly improved, and the high beta-glucosaccharase CLEAs residual relative enzyme activity can be maintained.

Description

technical field [0001] The invention belongs to the field of enzyme-catalyzed synthesis of salidroside, in particular to a process for catalyzing the synthesis of salidroside by β-glucosidase cross-linked aggregates (abbreviation: β-glucosidase CLEAs). Background technique [0002] Salidroside is a compound with special physiological functions. It has good effects in anti-cellular hypoxia, anti-tumor, and enhancing the body's immunity. Because the biosynthesis of salidroside has the advantages of high selectivity, mild reaction conditions, high product purity, and easy separation and purification of products, it has attracted widespread attention. [0003] In view of the fact that the reaction time of enzyme-catalyzed synthesis of salidroside is much shorter than that of plant tissue culture and cell culture, and the reaction process does not require expensive glycoside donors and catalysts, avoiding the cumbersome reaction steps of chemical synthesis. In addition, the react...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/44C12N9/42
Inventor 石贤爱薛原楷
Owner 福建东亮生物科技有限公司
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