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A method for synthesizing salidroside by attapulgite-loaded black Brin seed β-glucosidase cross-linked polymer

A technology of glucosidase and cross-linked polymer, which is applied in the field of synthesizing salidroside by attapulgite-loaded β-glucosidase cross-linked polymer of blackberry seeds, which can solve the problem of low concentration and high cost of salidroside , Low recovery rate of enzyme activity and other problems, to achieve the effect of improving the utilization rate of enzyme, low price, and improving the recovery rate of enzyme activity

Inactive Publication Date: 2017-09-29
HUAIYIN INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are not many β-glucosidases used for the synthesis of salidroside, and the concentration of the product salidroside is still low, the recovery rate of enzyme activity is low, and the cost is high, which cannot meet the needs of industrial production to maximize economic benefits. Therefore, the development of a more efficient β-glucosidase for the synthesis of salidroside and the use of this enzyme to explore the best reaction conditions and has great market application value

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: extracting black pulin seed β-glucosidase enzyme liquid 1

[0033] 1) Dehull and crush the black pudding seeds to obtain black pudding seed powder with a particle size of 100 mesh, then add 50g / L acetic acid-sodium acetate buffer solution with pH5.4, and then add Zhan black pudding seed powder The pectinase with a mass fraction of 1% was mixed evenly, incubated at a constant temperature of 40°C with shaking for 1 hour, the obtained solution was centrifuged, the supernatant was vacuum freeze-dried, and the powder was ground through a 60-mesh sieve to obtain a dry powder;

[0034]2) Step 1) Add 25 times the dry powder volume of phosphate buffer solution (concentration: 50 mM, pH: 7.4) as the extract, stir at 30 r / min for 1 h at 4°C, centrifuge, collect the extract, Add ammonium sulfate powder with 40% of the mass of the extract, stir for 30 min, centrifuge again, take the supernatant, add ammonium sulfate powder with a mass of 65% of the supernatant, stir at...

Embodiment 2

[0036] Embodiment 2: extracting black pulin seed β-glucosidase enzyme liquid 2

[0037] 1) Dehull and crush the black pudding seeds to obtain black pudding seed powder with a particle size of 100 mesh, then add 50g / L acetic acid-sodium acetate buffer solution with pH5.4, and then add Zhan black pudding seed powder The pectinase with a mass fraction of 2% was mixed evenly, incubated at a constant temperature of 40°C for 2 hours with shaking, the obtained solution was centrifuged, the supernatant was vacuum freeze-dried, and the powder was ground through a 60-mesh sieve to obtain a dry powder;

[0038] 2) Step 1) Add phosphate buffer solution (concentration: 50 mM, pH: 7.4) 50 times the volume of the dry powder to the dry powder as the extract, stir at 30 r / min for 1 h at 4°C, centrifuge, and collect the extract. Add ammonium sulfate powder with a mass of 40% of the extract, stir at 30r / min for 30 min, centrifuge again, take the supernatant, add ammonium sulfate powder with a ma...

Embodiment 3

[0040] Embodiment 3: extracting Heblin seed β-glucosidase enzyme liquid 3

[0041] 1) Dehull and crush the black pudding seeds to obtain black pudding seed powder with a particle size of 100 mesh, then add 50g / L acetic acid-sodium acetate buffer solution with pH5.4, and then add Zhan black pudding seed powder The pectinase with a mass fraction of 1.5% was mixed evenly, incubated at 40°C for 1.5 hours with constant temperature and shaking, the obtained solution was centrifuged, the supernatant was vacuum freeze-dried, and the powder was ground through a 60-mesh sieve to obtain a dry powder;

[0042] 2) Step 1) Add 35 times the dry powder volume of phosphate buffer solution (concentration: 50 mM, pH: 7.4) as the extract, stir at 30 r / min for 1 h at 4°C, centrifuge, collect the extract, Add ammonium sulfate powder with a mass of 40% of the extract, stir at 30r / min for 30 min, centrifuge again, take the supernatant, add ammonium sulfate powder with a mass of 65% of the supernatant...

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Abstract

The invention provides a method for synthesizing salidroside by attapulgite-loaded Heblin seed β-glucosidase cross-linked polymer, which is characterized in that: it comprises extracting Hei Bulin seed β-glucosidase enzyme solution, absorbing-cross-linked Synthesis of attapulgite-loaded β-glucosidase cross-linked polymer from Heblin seed by combined method and catalytic synthesis of three parts of salidroside. The present invention adopts the discarded Heibulin seed as the extraction raw material of β-glucosidase, which has large output, low price and easy acquisition. The activity recovery rate improves, and the tolerance to the environment is also significantly improved, and the product is easy to separate; the method of the present invention generally has a high enzyme activity recovery rate, good stability and reusability, and has a relatively high concentration of the product salidroside simultaneously. High advantages provide convenient conditions for large-scale production.

Description

technical field [0001] The invention belongs to the field of immobilized enzyme production, and relates to a method for catalyzing and synthesizing salidroside, in particular to a method for synthesizing salidroside by attapulgite-loaded β-glucosidase cross-linked polymer of black pudding seeds. Background technique [0002] Rhodiola generally grows in the plateaus of Europe and Asia. It is a perennial herb or subshrub of the Sedum family, and has the reputation of "plateau ginseng". There are 96 species of Rhodiola in the world, mainly distributed in East Asia, Central Asia, Siberia and North America. There are 73 species in my country, mainly distributed in the southwest, northwest, north and northeast regions. The main active ingredient of Rhodiola is salidroside, whose chemical name is p-hydroxyphenylethyl-β-D-glucoside. evaluation index. Salidroside has significant biological activity, and its effect is better than that of commonly used traditional Chinese medicines ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/44C12N11/14C12N9/42
CPCY02P20/50
Inventor 王朝宇朱春陆秀毕艳红杨荣玲赵祥杰
Owner HUAIYIN INSTITUTE OF TECHNOLOGY