CN-122012656-A - Preparation method and application of double-enzyme catalytic synthesis of gastrodin and gastrodin

Abstract

The invention discloses a preparation method for synthesizing gastrodin and gastrodin by double-enzyme catalysis and application thereof, belonging to the technical field of compound extraction. The technical problems to be solved are that the chemical synthesis process of gastrodin and gastrodin has large safety risk, more toxic and harmful reagent solvent residues, easy byproduct generation, limited yield and low purity, and the extraction and separation of traditional Chinese medicinal materials are time-consuming and labor-consuming, long in extraction time, low in extraction yield and purity, and the effective content of the medicine is low. The technical scheme is characterized in that immobilized cellulase and lipase are used as biocatalysts to catalyze the barrison glycoside compounds in the gastrodia elata extract to be converted into gastrodin and gastrodin, and the content of gastrodin and gastrodin in the gastrodia elata extract is improved, so that the drug effect is improved. The preparation process is environment-friendly, has high selectivity, can realize large-scale production, improves the extraction efficiency and the purity thereof, and reduces the cost.

Inventors

• CHEN CHEN
• YANG YINA
• CHEN KAI
• YANG XIAOZHOU
• HU HONGZHONG
• YANG HUI
• Gao Xianbing

Assignees

• 陕西理工大学
• 陕西百圣生物工程有限公司

Dates

Publication Date

20260512

Application Date

20260128

Claims (10)

1. 1. A preparation method for synthesizing gastrodin and gastrodin by double-enzyme catalysis is characterized by carrying out immobilization treatment on free cellulase and lipase to obtain immobilized cellulase and immobilized lipase, taking gastrodia elata as a raw material to obtain gastrodia elata extract freeze-dried powder, and synthesizing the balisen in the gastrodia elata extract into gastrodin and gastrodin under the catalysis of the immobilized cellulase and immobilized lipase by taking gastrodia elata extract freeze-dried powder as the raw material.
2. 2. The method of claim 1, wherein the method of preparing immobilized lipase comprises the steps of: (1.1) dissolving Fe 3 O 4 nano particles in water, adding dopamine hydrochloride after ultrasonic treatment to obtain a solution 1; (1.2) adjusting the pH of the solution 1 prepared in the step (1.1), and stirring to obtain a precipitate 1; (1.3) washing the precipitate 1 obtained in the step (1.2) for a plurality of times, and obtaining Fe 3 O 4 @dopamine hydrochloride through transparency to supernatant fluid, and freeze-drying for later use; (1.4) adding the Fe 3 O 4 @dopamine hydrochloride prepared in the step (1.3) and lipase into a phosphate buffer solution, and fixing for a period of time to obtain a precipitate 2; (1.5) washing and freeze-drying the precipitate 2 obtained in the step (1.4) for later use.
3. 3. The method of claim 1, wherein the method of preparing immobilized cellulase comprises the steps of: Adding Fe 2 O 3 nano particles into a cellulase solution containing polyvinyl alcohol, stirring, quickly freezing, thawing, repeatedly freezing and thawing for many times, adding NaCl for sedimentation, and then carrying out adsorption separation, cleaning, precipitation and freeze drying for later use.
4. 4. The preparation method of the gastrodia elata extract freeze-dried powder according to claim 1, which is characterized by comprising the following steps of: (3.1) selecting fresh gastrodia elata, cleaning, peeling, slicing, steaming, drying and crushing to obtain gastrodia elata powder for later use; (3.2) adding water into the gastrodia elata powder obtained in the step (3.1), carrying out reflux extraction, standing, centrifuging to obtain a supernatant, extracting the residual filter residues repeatedly, and combining the filtrates to obtain a solution A; (3.3) adding active carbon into the solution A in the step (3.2), stirring, standing, filtering and decoloring to obtain filtrate B; And (3.4) freezing and solidifying the filtrate B, and then freeze-drying to obtain the gastrodia elata extract freeze-dried powder.
5. 5. The preparation method of claim 4, wherein the mass-to-volume ratio of the gastrodia elata powder to the water in the step (3.2) is 1g:30-44mL, and the reflux extraction temperature is 90-110 ℃.
6. 6. The method according to claim 4, wherein the reflux extraction in the step (3.2) is performed for 1 to 3 hours each time, and the centrifugation is performed at 2 to 8 ℃.
7. 7. The preparation method of claim 1, wherein the synthesis of gastrodin and gastrodin from the barrison glycoside in the gastrodia elata extract comprises the following steps: (4.1) adding water into immobilized lipase, immobilized cellulase and gastrodia elata extract freeze-dried powder, stirring, adjusting pH, heating and oscillating for a period of time, and then carrying out constant-temperature reaction to obtain a material C; And (4.2) centrifuging the material C, and taking supernatant for recovery to obtain the product.
8. 8. The method according to claim 7, wherein the amount ratio of the gastrodia elata extract freeze-dried powder, the immobilized cellulase and the immobilized lipase in the step (4.1) is 100:2.0-5.0:1.0-2.0.
9. 9. Gastrodin and gastrodin prepared by the method of any one of claims 1-8.
10. 10. Use of gastrodin and gastrodin according to claim 9 for the preparation of a medicament for the treatment of neurological, cardiovascular or immune disorders.

Description

Preparation method and application of double-enzyme catalytic synthesis of gastrodin and gastrodin Technical Field The invention belongs to the technical field of compound extraction, and particularly relates to a preparation method for synthesizing gastrodin and gastrodin by double-enzyme catalysis and application thereof. Background The rhizoma Gastrodiae contains polyphenol compound, polysaccharide, protein, amino acid and microelements. The chemical components include gastrodin, gastrodin (p-hydroxybenzyl alcohol), p-hydroxybenzaldehyde, balrison glycoside A, balrison glycoside B, balrison glycoside C, balrison glycoside D, balrison glycoside E, etc., wherein the gastrodin and the gastrodin are chemical components which are required to be detected in Chinese pharmacopoeia, and the sum of the two chemical components is more than or equal to 0.25%. Gastrodin has various pharmacological activities, has remarkable potential in neuroprotection, antiepileptic, antiinflammatory, antidepressant, antitumor, and blood sugar regulating effects, and can be used for resisting epilepsia, neuroprotection, tranquilizing, relieving inflammation, resisting oxidation, scavenging free radicals, neuroprotection, and inhibiting tumor cell growth. After being absorbed into blood, gastrodin is decomposed into gastrodin aglycone and glucose, the pharmacological actions of the gastrodin and the gastrodin aglycone are similar, the main substance basis for playing the pharmacological actions of the gastrodin and the gastrodin is the same, and the gastrodin has wide clinical application in the aspects of treating diseases of nervous system, cardiovascular system, immune system and the like, and has no obvious toxic or side effect. The gastrodin has a chemical structure of p-hydroxymethylphenyl beta-D-glucopyranoside, and is formed by connecting a molecule of beta-D-glucopyranoside with a molecule of gastrodin through glycosidic bonds. The gastrodin has a chemical structure of p-hydroxy benzyl alcohol, and comprises a benzene ring (C 6H5) with two hydroxyl (-OH) groups. Besides the chemical components regulated by the 2 Chinese pharmacopoeias, the gastrodia elata also has a high research value for a plurality of active components such as the barrison glycoside A, the barrison glycoside B, the barrison glycoside C, the barrison glycoside D, the barrison glycoside E and the like. The balrison glycoside A belongs to glycoside compounds, and consists of one molecule of citric acid at 1,2 and 3 positions and three molecules of gastrodin through ester bonds, the balrison glycoside B consists of one molecule of citric acid at 1,2 positions and two molecules of gastrodin through ester bonds, the molecular formula of the balrison glycoside C is the same as that of the balrison glycoside B, the difference is that the balrison glycoside B contains two glucose molecules, the balrison glycoside C contains two different sugars of glucose and rhamnose, the glycosidic bonds of the two glucose molecules are not completely the same, the balrison glycoside D is obtained by esterifying one molecule of citric acid and two molecules of gastrodin, and the balrison glycoside E consists of one molecule of citric acid at 1 position and one molecule of gastrodin through ester bonds. The balison glycoside compounds are usually formed by condensing one molecule of citric acid and a polymolecular gastrodin group, belong to polyphenol or polyphenol glycoside compounds, and can be mutually converted with gastrodin. According to the characteristic that the chemical structure of the balison substance contains glycosidic bonds and/or ester bonds, the balison substance has the characteristic of unstable chemical property, and is easy to damage and inactivate in the processing process. At present, gastrodin and gastrodin are mainly prepared by extraction, separation and chemical synthesis from traditional Chinese medicines, but the extraction, separation and chemical synthesis have disadvantages. In the process of extracting and separating gastrodin and gastrodin from gastrodia elata, extraction methods such as decoction or soaking are often adopted, and organic solvents are often used in the extraction process, so that the time and the labor are wasted, the extraction time is long, the extraction yield is low, the extraction waste liquid is large, the waste liquid treatment cost is high, and the obtained extract contains not only effective components but also other ineffective components. Gastrodine and gastrodin can not be completely separated out as effective active ingredients, and even some medicinal materials are boiled or soaked for too long to dissipate the efficacy, so that the effects of treating hypertension and protecting blood vessels are obviously reduced, the dosage of the medicaments is large, the patients take the medicaments, and the application limitation on the market is large. The gastrodin and the gastrodin are respectively synthesized and prepare