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Sulfonated polystyrene microsphere immobilized alkaline protease and preparation method thereof

A technology of sulfonated polystyrene and polystyrene microspheres, applied in biochemical equipment and methods, immobilized on/in organic carriers, enzymes, etc., can solve the problem of unloaded sulfonated polystyrene microspheres Alkaline protease and other problems, to achieve the effects of good dispersibility, strong electronegativity, high specific surface area, and simple and convenient method

Inactive Publication Date: 2019-01-11
SOUTH CHINA UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no report on the loading of alkaline protease on sulfonated polystyrene microspheres.

Method used

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  • Sulfonated polystyrene microsphere immobilized alkaline protease and preparation method thereof
  • Sulfonated polystyrene microsphere immobilized alkaline protease and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Prepare a 4.5 mg / mL Bacillus subtilisin solution with distilled water, and add the nano-sulfonated polystyrene microsphere solution that has been uniformly dispersed by ultrasound to make the final concentration of the microspheres 1 mg / mL. At room temperature, immobilize with magnetic stirring for 2 minutes, centrifuge and wash with water three times to obtain a precipitate of enzyme immobilized on sulfonated polystyrene microspheres. Collect the combined supernatant, measure the protein content according to the Coomassie Brilliant Blue method, and calculate the enzyme load; collect the precipitate, measure the enzyme activity according to the national industry standard SB / T 10317 protease activity assay, and calculate the enzyme activity recovery rate, the calculation formula is as follows (1 ), (2) shown. The activity recovery rate of the immobilized enzyme was 91.59%, and the loading capacity was 273.36 mg / g.

[0030] (1) Enzyme activity recovery (%) = immobilized ...

Embodiment 2

[0036] Prepare a 4.5 mg / mL Bacillus subtilisin solution with distilled water, and add the nano-sulfonated polystyrene microsphere solution that has been uniformly dispersed by ultrasound to make the final concentration of the microspheres 1 mg / mL. At room temperature, immobilized by magnetic stirring for 5 min, centrifuged, and washed with water three times to obtain sulfonated polystyrene microspheres immobilized enzyme, the activity recovery rate was 82.55%, and the loading capacity was 288.6 mg / g.

Embodiment 3

[0038] Prepare a 4.5 mg / mL Bacillus subtilisin solution with distilled water, and add the nano-sulfonated polystyrene microsphere solution that has been uniformly dispersed by ultrasound to make the final concentration of the microspheres 1 mg / mL. At room temperature, immobilized by magnetic stirring for 10 min, centrifuged, washed with water three times to obtain sulfonated polystyrene microspheres immobilized enzyme, the activity recovery rate was 80.49%, and the loading capacity was 306.38 mg / g.

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Abstract

The invention belongs to the technical field of immobilized enzymes and discloses sulfonated polystyrene microsphere immobilized alkaline protease and a preparation method thereof. The preparation method comprises the following steps: ultrasonically dispersing sulfonated polystyrene microspheres into distilled water; then adding into an alkaline protease solution and carrying out adsorption and immobilization reaction; and separating and washing a product to obtain the sulfonated polystyrene microsphere immobilized alkaline protease. The sulfonated polystyrene microsphere immobilized alkalineprotease prepared by the invention not only has high activity and recycling rate and a high loading amount, but also has a simple and rapid immobilization process; and the problem that the effect of atraditional immobilized enzyme is not ideal is solved.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, in particular to a sulfonated polystyrene microsphere immobilized alkaline protease and a preparation method thereof. Background technique [0002] Protease is the most widely used type of enzyme in industrial enzymes, accounting for about 60% of the total amount of enzymes, and alkaline protease accounts for more than 50% of the sales of protease market. It has played a huge role in the food industry. Alkaline protease refers to an enzyme that hydrolyzes protein peptide bonds in the alkaline pH range. As a biocatalyst, it has incomparable advantages over ordinary chemical catalysts: high efficiency, specificity, and mild reaction conditions , low pollution, wide range of sources. However, in practical applications, enzymes have higher requirements on environmental conditions, and enzymes are more sensitive to various physical or chemical factors, such as strong acid, strong alkali, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12N11/04C12N9/56
CPCC12N9/54C12N11/04C12N11/08C12Y304/21062
Inventor 肖凯军杨小翠谌凯
Owner SOUTH CHINA UNIV OF TECH
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