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Synthesis of salidroside catalyzed by β-glucosidase cross-linked aggregates

A technology of glucosidase and salidroside, which is applied in the field of enzyme-catalyzed synthesis of salidroside, which can solve the problems of low concentration, inability to meet the needs of maximizing economic benefits, and low recovery rate of enzyme activity, so as to reduce the usage amount , High residual relative enzyme activity, high enzyme catalytic efficiency

Active Publication Date: 2016-02-24
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the concentration of the product salidroside is still low, which cannot meet the demand for maximizing economic benefits in industrialized production, and the recovery rate of enzyme activity is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] β-glucosidase CLEAs catalyzes the process of synthesizing salidroside, including the preparation of β-glucosidase CLEAs and the synthesis of salidroside;

[0022] The preparation stage of β-glucosidase CLEAs includes the following steps: (1) Add 2.2ml of 1,4-dioxane to 1ml of β-glucosidase solution containing 1.5mg / ml of β-glucosidase (1, The volume ratio of 4-dioxane to β-glucosidase solution is 2.2:1), shake; (2) add glutaraldehyde to the final concentration of 40mM, react for 10min, and carry out cross-linking; (3) Sodium borohydride (NaBH 4 ), to NaBH 4 The final concentration of 20mM 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 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%.

[0023] The synthesi...

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 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), shake; (2) add glutaraldehyde to a final concentration of 38mM, react for 12min, and carry out cross-linking; (3) Sodium borohydride (NaBH 4 ), to NaBH 4 The final concentration of 25mM was reduced, after shaking, centrifuged at 500rpm for 5min, and the precipitate was collected, which was β-glucosidase CLEAs; the reaction temperature was controlled at 6°C during the whole reaction process, and the pH was 5.2; the precipitate was collected to measure β-glucosidase CLEAs. The recovery rate of the enzyme activity of glucosidase CLEAs was 81.71%.

[0030] The synthesis stage of ...

Embodiment 3

[0032] β-glucosidase CLEAs catalyzes the process of synthesizing salidroside, including the preparation of β-glucosidase CLEAs and the synthesis of salidroside;

[0033] The preparation stage of β-glucosidase CLEAs includes the following steps: (1) Add 2.0ml of 1,4-dioxane to 1ml of β-glucosidase solution containing 1.5mg / ml of 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 β-glucosidase CLEAs was reduced to 23mM, after shaking, centrifuged at 300rpm for 10min, and the precipitate was collected, which was β-glucosidase CLEAs; the reaction temperature during the whole reaction process was controlled at 4~6°C, and the pH was 5.0; the precipitate was collected and measured The recovery rate of β-glucosidase CLEAs was 80.59%.

[0034] The synthesis stage of salidros...

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Abstract

The invention provides a process for synthesizing salidroside under catalysis of beta-glucosidase cross-linked aggregates. Reaction conditions for a synthetic stage of salidroside are that a reaction system is 1-butyl-3-methylimidazole hexafluorophosphate (C4MIm.PF6) / 1,4-dioxane / buffer solution; unit enzyme activity of the beta-glucosidase cross-linked aggregates is 1.3-1.6 U / ml; a volume ratio of C4MIm.PF6 to 1,4-dioxane is 2:3-3:2; a volume fraction of the buffer solution is 12-18%; a reaction temperature is 48-52 DEG C; a rotation speed is 245-255 rpm; and a reaction time is 5-9 d. The process can greatly increase the concentration of the product salidroside, and can keep relatively high residual relative enzyme activity of the beta-glucosidase cross-linked aggregates.

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 Patents(China)
IPC IPC(8): C12P19/44C12N9/42
Inventor 石贤爱薛原楷
Owner FUZHOU UNIV
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