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Method for enzymatic synthesis of pinoresinol active substances

A technology for enzymatic synthesis and active substances, which is applied in the fields of enzymatic synthesis of pinoresinol active substances and synthesis of pinoresinol active substances, can solve problems such as process pollution, and achieve the effects of low cost, environmental protection and high yield.

Active Publication Date: 2014-05-07
CHINA UNIV OF PETROLEUM (BEIJING)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some data show that in the process of laccase catalysis at home and abroad, because the problem of solvent use has not been solved, there is a big shortage in the efficient use of this type of biocatalyst, and there is still serious process pollution.

Method used

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  • Method for enzymatic synthesis of pinoresinol active substances
  • Method for enzymatic synthesis of pinoresinol active substances

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The present embodiment provides a kind of preparation method of pinoresinol active material, and it comprises the following steps:

[0033] Sodium hydrogen phosphate buffer solution (concentration is 0.1mol / L, pH value is 6.8) and acetone are mixed according to the volume ratio of 1:1 to obtain mixed solution, in this mixed solution, the volume content of phosphate buffer solution is 50%;

[0034] Add 1mmol pinoresinol substrate coniferyl alcohol to 25mL mixed solution;

[0035] Add 1 μg anacardinase and react at room temperature for 72 hours;

[0036] After having reacted, add appropriate amount of methyl alcohol and sodium tartrate (methanol and water miscible organic solvent equimolar quantity, sodium tartrate is 1 / 2nd of the molar quantity of phosphate buffer saline) in reaction product, utilize ethyl acetate to reaction product then Extract, wash with saturated sodium chloride, dry over anhydrous sodium sulfate and concentrate, then recrystallize with methanol, c...

Embodiment 2

[0040] The present embodiment provides a kind of preparation method of pinoresinol active material, and it comprises the following steps:

[0041] Potassium hydrogen phosphate buffer solution (concentration is 0.1mol / L, pH value is 6.8) and acetone are mixed according to the volume ratio of 4:1 to obtain mixed solution, in this mixed solution, the volume content of phosphate buffer solution is 80%;

[0042] 1mmol pinoresinol substrate isoeugenol was added to 50mL mixed solution;

[0043] Add 10 μg anacardinase and react at room temperature for 72 hours;

[0044] After having reacted, add appropriate amount of methyl alcohol and sodium tartrate (methanol and water miscible organic solvent equimolar quantity, sodium tartrate is 1 / 2nd of the molar quantity of phosphate buffer saline) in reaction product, utilize ethyl acetate to reaction product then Extract, wash with saturated sodium chloride, dry over anhydrous sodium sulfate and concentrate, then recrystallize with methanol...

Embodiment 3

[0048] The present embodiment provides a kind of preparation method of pinoresinol active material, and it comprises the following steps:

[0049] Disodium hydrogen phosphate and potassium dihydrogen phosphate buffer solution (concentration is 0.1mol / L, pH value is 6.8) and ethanol are mixed according to the volume ratio of 1: 1 to obtain mixed solution, in this mixed solution, the phosphate buffer saline The volume content is 50%;

[0050] Add 1mmol pinoresinol substrate coniferyl alcohol to 50mL mixed solution;

[0051] Add 15 μg anacardinase and react at room temperature for 72 hours;

[0052] After having reacted, add appropriate amount of methyl alcohol and sodium tartrate (methanol and water miscible organic solvent equimolar quantity, sodium tartrate is 1 / 2nd of the molar quantity of phosphate buffer saline) in reaction product, utilize ethyl acetate to reaction product then Extraction, washing with saturated sodium chloride, drying over anhydrous sodium sulfate and c...

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Abstract

The invention relates to a method for enzymatic synthesis of pinoresinol active substances, which includes the steps: mixing water or phosphate buffer solution with water-soluble organic solvent into mixed solution; adding pinoresinol substrate into the mixed solution; adding Rhus Laccase to react at the temperature of 20-45 DEG C for 48-96 hours; after reaction, adding a proper amount of methanol and sodium tartrate into reaction product, subjecting the reaction product to ethyl acetate extraction, anhydrous sodium sulfate drying and concentration, and performing methanol recrystallization to obtain crystal or performing silica-gel column chromatography, concentration and purification to obtain chromatographic product; and subjecting the crystal or chromatographic product to nitrogen blowing or freeze-drying to obtain pinoresinol active substances. The method has moderate conditions, reaction is environment-friendly, simple, quick and low-cost, and the method has broad application prospect in the fields of preparing similar biological medicinal products, active complex compounds, and the like.

Description

technical field [0001] The invention relates to a method for synthesizing an active substance of pinoresinol, in particular to an enzymatic synthesis method for an active substance of pinoresinol, and belongs to the technical field of synthesis of active compounds. Background technique [0002] Pinoresinol is one of the active ingredients in Eucommia ulmoides, a specialty Chinese herbal medicine in China. It has been used in my country for thousands of years. The earliest written records can be traced back to the book "Shen Nong's Herbal Classic" written in the early Eastern Han Dynasty (about 25 AD). . [0003] Modern studies have shown that pinoresinol and its derivatives such as monomethyl ester, dimethyl ester and β-D-glucoside all have the functions of inhibiting the activity of cyclic adenylate phosphodiesterase, as well as antihypertensive and tumor inhibiting functions. The pharmacological functions of Eucommia mainly come from active ingredients such as pinoresinol....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/00C12P7/22
Inventor 万云洋杜予民
Owner CHINA UNIV OF PETROLEUM (BEIJING)
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