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Method for preparing ginsenoside-Rd through immobilization of cellulase and enzymolysis of ginsenoside-Rb1 by virtue of covalent cross-linking process

A technology of ginsenoside and cellulase, applied in the direction of fermentation, can solve the problems of weak enzyme binding force, easy inactivation contact surface area, etc., and achieve the effects of high selectivity, convenient continuous and automatic operation, and saving production cost.

Active Publication Date: 2017-03-22
LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the above-mentioned prior art, only the embedding method and the cross-linking method are used, and the enzyme has the disadvantages of weak binding force, easy inactivation and small contact surface area.

Method used

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  • Method for preparing ginsenoside-Rd through immobilization of cellulase and enzymolysis of ginsenoside-Rb1 by virtue of covalent cross-linking process
  • Method for preparing ginsenoside-Rd through immobilization of cellulase and enzymolysis of ginsenoside-Rb1 by virtue of covalent cross-linking process
  • Method for preparing ginsenoside-Rd through immobilization of cellulase and enzymolysis of ginsenoside-Rb1 by virtue of covalent cross-linking process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Covalent immobilization process of cellulase.

[0023] Weigh 0.25g of carrageenan (purchased from Beijing Suolaibao Company) and add it to 25ml of distilled water at a temperature of 65°C. Stir with a magnetic stirrer at a speed of 300rpm for 1h to obtain a carrageenan suspension; take 4°C 50ml of 2% KCl solution was placed in a beaker, stirred on a magnetic stirrer at a speed of 300rpm, and the carrageenan suspension was sucked up with a 5ml syringe, and slowly dropped into it (the drop distance was fixed at 6 cm during the dropping process, and the dropping distance was 6 cm. The speed is 80 drops / min), and transparent hemispherical gel beads are formed instantaneously, and placed in 2% KCl solution for 3 hours to make gel beads with a concentration of 1% carrageenan; take 1g of carrageenan gel beads, add 10ml concentration For 1% polyethyleneimine solution, add 10% NaOH solution dropwise to adjust the pH value to 10, put it on a roller mixer for mixing and amination ...

Embodiment 2

[0025] Covalent immobilization process of cellulase.

[0026] Weigh 0.5g of carrageenan and add it to 25ml of distilled water. Stir with a magnetic stirrer at a temperature of 65°C at a speed of 300rpm to obtain a suspension of carrageenan; take 50ml of 2% KCl solution at 4°C and place it in a beaker , stirred on a magnetic stirrer with a rotation speed of 300rpm, and sucked the carrageenan suspension with a 5ml syringe, and slowly dropped it into it (during the dropping process, the fixed drop distance was 6 cm, and the dropping speed was 80 drops / min), and the Transparent hemispherical gel beads, and placed in 2% KCl solution for 3 hours to make 1% carrageenan gel beads; take 1 g of carrageenan gel beads, add 10 ml of 0.5% polyethyleneimine solution , add 10% NaOH solution dropwise to adjust the pH value to 8, place it on a roller mixer for amination for 3 hours, wash with distilled water 3 times, and clean the residual polyethyleneimine; then add it to 10 ml of 3% glutarald...

Embodiment 3

[0031] Conversion of Ginsenoside-Rb from Panaxadiol Group Saponins by Immobilized Enzyme 1 .

[0032] 1. Preparation of saponin samples of American ginsengdiol group.

[0033] American ginseng dry root and rhizome coarse powder are decocted and extracted with water, the residue is filtered off, the water is concentrated under reduced pressure to the concentration of American ginseng, and then separated on D 101 macroporous adsorption resin. The amount of macroporous adsorption resin is 1.5 times the concentrated solution. Adsorb at 55°C for 12 h, wash with water until there is no sweetness, then desorb with 30% and 65% ethanol solutions in sequence, collect 65% of the eluent, spin the solvent under reduced pressure, and further dry under reduced pressure to obtain American ginseng diol Saponin dry powder.

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Abstract

The invention belongs to the technical field of immobilization of enzymes and particularly relates to a method for preparing ginsenoside-Rd through immobilization of cellulase and enzymolysis of ginsenoside-Rb1 by virtue of a covalent cross-linking process. An immobilized enzyme is prepared through three steps of polyethyleneimine amination, glutaraldehyde cross-linking and enzyme immobilization. Compared with a manner of simply utilizing a free enzyme, the method has the advantages of recyclability, high selectivity, no pollution and the like. Compared with other enzyme immobilization methods, the method has many advantages that the bonding is firm, the operation is stable and the repeated utilization is realized, and the method is more suitable for industrial production.

Description

technical field [0001] The invention belongs to the technical field of enzyme immobilization, in particular to a covalent cross-linking immobilization of cellulase and enzymolysis of ginsenoside-Rb 1 Method for preparing ginsenoside-Rd. Background technique [0002] Ginsenoside-Rd belongs to protopanaxadiol type saponins in tetracyclic triterpene dammarane type. Studies have shown that protopanaxadiol-type saponins and protopanaxatriol-type saponins, the main metabolic pathway in the body is to generate secondary glycosides and aglycones with better pharmacological activity through desugaring reactions, and intestinal enzymes convert protopanaxadiol Ginsenoside-Rb 1 Metabolized to -Rd. Therefore, ginsenoside-Rd is one of the important forms absorbed and utilized by the intestinal tract after the metabolism of protopanaxadiol saponins. [0003] Ginsenoside-Rd has the functions of protecting the cardiovascular system, anti-tumor, immune regulation, and neuroprotection. At...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P33/20
CPCC12P33/005C12P33/20
Inventor 窦德强冉小库默罕默德·艾尔赛义德·阿里·哈桑袁颖栾晓宁
Owner LIAONING UNIV OF TRADITIONAL CHINESE MEDICINE
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