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Method for preparing cycloastragenol by catalyzing astragaloside through co-immobilized double enzymes

A technology of astragaloside IV and cycloastragenol, which is applied in the field of catalysis and can solve the problems of limiting production efficiency, increasing production costs, and the inability to realize recycling and reuse of free enzymes.

Active Publication Date: 2020-10-30
WEIHAI BAIHE BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since the separation of intermediates is involved, and the free enzyme cannot be recycled and reused after use, the production cost is increased and the production efficiency is greatly limited.

Method used

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  • Method for preparing cycloastragenol by catalyzing astragaloside through co-immobilized double enzymes
  • Method for preparing cycloastragenol by catalyzing astragaloside through co-immobilized double enzymes
  • Method for preparing cycloastragenol by catalyzing astragaloside through co-immobilized double enzymes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Prepare a phosphate buffer solution by mixing disodium hydrogen phosphate with a concentration of 0.2mol / L and a volume of 19.15mL and citric acid with a concentration of 0.1mol / L and a volume of 0.85mL to prepare a phosphate buffer solution with a concentration of 1mg / mL and a volume of 60μL β-glucosidase was added to 1 mL of phosphate buffered saline solution (take part of the prepared phosphate buffered saline solution). Then add Fe at a concentration of 5 mg / mL in a volume of 100 μL 3 o 4 Mix the solution, then add a copper chloride solution with a concentration of 180mM and a volume of 100μL for reaction, let it stand at room temperature for 24 hours, collect the precipitate with a magnet, and mix the collected precipitate with 1mg / mL xylosidase and phosphate buffer solution (same composition as the above-mentioned phosphate buffer solution) for the reaction, the volumes of xylosidase and phosphate buffer solution were 60 μL and 1.2 mL respectively. After standin...

Embodiment 2

[0048] Prepare a phosphate buffer solution by mixing disodium hydrogen phosphate with a concentration of 0.15 mol / L and a volume of 23.15 mL and citric acid with a concentration of 0.07 mol / L and a volume of 1.85 mL to prepare a phosphate buffer solution with a concentration of 0.5 mg / mL and a volume of 40 μL β-glucosidase was added to 0.8mL phosphate buffer solution (take part of the prepared phosphate buffer solution). Then add Fe at a concentration of 3 mg / mL in a volume of 80 μL 3 o 4 Mix the solution evenly, then add a copper chloride solution with a concentration of 150mM and a volume of 80μL for reaction, let it stand at room temperature for 18 hours, collect the precipitate with a magnet, and mix the collected precipitate with 0.5mg / mL xylosidase, phosphate buffered saline Solution (same composition as the above-mentioned phosphate buffer solution) was reacted, and the volumes of xylosidase and phosphate buffer solution were 40 μL and 1.0 mL, respectively. After stan...

Embodiment 3

[0052] Prepare a phosphate buffer solution by mixing disodium hydrogen phosphate with a concentration of 0.2mol / L and a volume of 19.15mL and citric acid with a concentration of 0.1mol / L and a volume of 0.85mL to prepare a phosphate buffer solution with a concentration of 2.5mg / mL and a volume of 80μL β-glucosidase was added to 1.6mL phosphate buffer solution (take part of the prepared phosphate buffer solution). Then add Fe at a concentration of 8 mg / mL in a volume of 120 μL 3 o 4 The solution was mixed evenly, and then a copper chloride solution with a concentration of 250 mM and a volume of 120 μL was added for reaction. After standing at room temperature for 24 hours, the precipitate was collected with a magnet, and the collected precipitate was mixed with 2.5 mg / mL xylosidase, phosphate buffered saline Solution (same composition as the above-mentioned phosphate buffer solution) was reacted, and the volumes of xylosidase and phosphate buffer solution were 80 μL and 2 mL, ...

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Abstract

The invention belongs to the technical field of catalysis, and provides a method for preparing cycloastragenol by catalyzing astragaloside through co-immobilized double enzymes. The method comprises the following steps of: carrying out a reaction among beta-glucosidase, a phosphate buffer solution, a Fe3O4 solution, a copper chloride solution and xylosidase to obtain the co-immobilized double enzymes; and carrying out a catalytic reaction by using co-immobilized double enzymes and astragaloside to obtain cycloastragenol. The method provided by the invention overcomes the problem of extractionand separation of intermediates in a process for preparing cycloastragenol by adopting a two-step reaction in the prior art, and meanwhile, the co-immobilized double enzymes can be recycled, so that the production cost is reduced, the process is simple, economic and economical, and the method is very suitable for large-scale industrial production. The conversion rate of the co-immobilized double enzymes to astragaloside basically reaches 100%, and the substrate astragaloside is completely converted into cycloastragenol. The purity of the cycloastragenol product obtained by the method can reach78.3% or above.

Description

technical field [0001] The invention relates to the technical field of catalysis, in particular to a method for preparing cycloastragenol by co-immobilizing double enzymes to catalyze astragaloside IV. Background technique [0002] Multienzyme processes play an important role in biotransformation and biomedical engineering, and are considered as alternatives for the production of pharmaceuticals, biofuels, and fine chemicals. Initially, only a single enzyme was purified from microorganisms for in vitro biotransformation reactions. At present, the multi-enzyme cascade reaction assembled in the form of a multi-enzyme complex integrates the transformation process of multiple enzymes, which can eliminate the problem of difficult separation and purification of reaction intermediates, thereby significantly reducing costs and saving energy. Fewer unit operations, shorter cycle times, smaller reactor volumes, and better space-time yields also lead to better economics, enabling sust...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/18C12N9/24C12N9/42C12P33/20C12P33/06
CPCC12N11/14C12N11/18C12N9/2445C12N9/2402C12Y302/01021C12P33/20C12P33/06
Inventor 袁其朋梁浩魏斌程磊雨高慧玲石晔刘旭东张军杰
Owner WEIHAI BAIHE BIOTECH
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