Method for preparing salidroside with enzymatic method

A salidroside, enzymatic preparation technology, applied in the field of enzymatic preparation of salidroside, can solve the problems of not much improvement in synthesis results, poor enzyme activity and stability, disadvantages, etc., to achieve improved thermal stability, process The effect of mild conditions and low volatility

Active Publication Date: 2016-12-14
SHANXI UNIV
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Nevertheless, in the salidroside biosynthesis reaction, since the enzyme, substrate and product are insoluble in organic solvents, and the activit

Method used

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  • Method for preparing salidroside with enzymatic method
  • Method for preparing salidroside with enzymatic method
  • Method for preparing salidroside with enzymatic method

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Embodiment 1

[0029] A. Synthesis of deep eutectic solvent: Accurately weigh 13.97g of choline chloride, measure 14.56mL of glycerin, mix them into a 500mL round bottom flask, heat and stir in a collector magnetic stirrer at a temperature of 80-100°C , the speed is 300rpm. After 10 h of reaction, a uniform transparent liquid was formed, and about 30 mL of DES was obtained.

[0030] B. Preparation of β-D-glucosidase immobilized on chitosan microspheres: Weigh 0.5 g of chitosan and dissolve it in 100 mL of 2% glacial acetic acid to prepare chitosan colloidal solution, and ultrasonically remove air bubbles. Use a syringe to draw an appropriate amount of the above-mentioned chitosan solution, drop it into the coagulation liquid, the composition of the coagulation liquid is 7.5% NaOH solution / 95% ethanol (v / v) = 4:1, and form microspheres with a size of about 1.5mm. , added to 3% glutaraldehyde solution, oscillating and cross-linking at 50°C for 2h, washing away excess glutaraldehyde, adding β-...

Embodiment 2

[0033] A. With embodiment 1.

[0034] B. With embodiment 1.

[0035] C. Enzyme-catalyzed synthesis of salidroside: Accurately weigh 1.8g of anhydrous D-glucose and 11.06g of tyrosol, and stir to fully dissolve in 180mL DES, then add 20mL of citric acid at pH 5.8 pre-incubated at 45°C in sequence - Disodium hydrogen phosphate buffer solution, 10000U chitosan microsphere immobilized β-D glucosidase, mix well. The above system was heated to 45° C., kept warm and stirred for 120 h. After the reaction, the reaction solution was filtered out and diluted with an equal volume of methanol. The conversion rate of the substrate (glucose meter) was detected by HPLC to reach 32.9%.

Embodiment 3

[0037] A. With embodiment 1.

[0038] B. With embodiment 1.

[0039] C. Enzyme-catalyzed synthesis of salidroside: Accurately weigh 1.8g of anhydrous D-glucose and 12.44g of tyrosol, stir to dissolve them in 140mL of DES, then add 60mL of citric acid at pH5.8 pre-incubated at 45°C in sequence - Disodium hydrogen phosphate buffer solution, 7000U chitosan microsphere immobilized β-D glucosidase, mix well. The above system was heated to 50° C., kept warm and stirred for 120 h. After the reaction, the reaction liquid was filtered out and diluted with an equal volume of methanol. The conversion rate of the substrate (glucose meter) was detected by HPLC to reach 31.8%.

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Abstract

The invention provides a method for preparing salidroside with an enzymatic method. The method includes the specific steps that deep-eutectic solvents (DESs) synthesized by heating and stirring choline chloride and glycerin serve as reaction solvents, and the reaction solvents are biologically catalyzed and glycosylated into salidroside through beta-D-glucosidase, wherein the conversion rate of a substrate can be 30% or above (by glucose). According to the method, the adopted DESs are green and nontoxic, the solubility of the reaction substrate is high, and good biocompatibility is achieved. The preparing method has the advantages of being easy to operate, efficient, environmentally friendly, mild in condition, low in cost and the like.

Description

technical field [0001] The invention relates to a method for synthesizing salidroside, in particular to a method for enzymatically preparing salidroside. Background technique [0002] Rhodiola rosea (Rhodiola rosea L.), a perennial herb, grows in the alpine and pollution-free zone at an altitude of 1800-2500 meters. Its growth environment is harsh, so it has strong vitality and special adaptability. It can be used as medicine, it can invigorate qi and clear the lungs, improve intelligence and nourish the heart. It is a traditional Chinese medicine with a wide range of functions. It also has a great beauty effect, and can be used as skin care products and can also be eaten. [0003] Salidroside, molecular formula C 14 h 20 o 7 , the structural formula: [0004] [0005] It is the main active ingredient in Rhodiola rosea. It has the functions of anti-hypoxia, anti-cold, anti-fatigue, anti-radiation, delaying the aging of the body, and preventing senile diseases. It is v...

Claims

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Application Information

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IPC IPC(8): C12P19/44
CPCC12P19/44
Inventor 张立伟程启斌
Owner SHANXI UNIV
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