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Enzyme-inorganic crystal compound microsphere and preparation method thereof

A technology of inorganic crystals and composite microspheres, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of high preparation cost, complicated preparation method, low yield of immobilized enzyme activity, etc., and achieve simple operation and low cost , the effect of stability improvement

Inactive Publication Date: 2014-10-01
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing single-enzyme and multi-enzyme immobilization methods have limitations, such as complex preparation methods, low activity yield of immobilized enzymes, and high preparation costs. High-efficiency, low-cost methods are important

Method used

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  • Enzyme-inorganic crystal compound microsphere and preparation method thereof
  • Enzyme-inorganic crystal compound microsphere and preparation method thereof
  • Enzyme-inorganic crystal compound microsphere and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of candida antarctica lipase B (CALB)-copper phosphate crystal composite microsphere

[0045] 1. Prepare CALB PBS buffer (pH=7.4) with an enzyme concentration of 0.5 mg / mL and 200 mM copper sulfate pentahydrate aqueous solution.

[0046] 2. Take 5mL of the enzyme solution in step 1 into a glass vial, add 0.05mL of the copper sulfate solution in step 1, mix well, and put it in a 25°C incubator for 24h.

[0047] 3. After filtering the mixture obtained in step 2, the precipitate was collected, washed 3 times with deionized water, filtered again to collect the precipitate, and freeze-dried for 5 hours to obtain copper phosphate crystal-Candida antarctica lipase B composite microspheres.

[0048] The scanning electron micrograph of the composite microsphere prepared in this embodiment is as follows figure 2 shown by figure 2 It can be seen that the particle size of the composite microsphere is about 10 μm. The enzyme loading in the composit...

Embodiment 2-4

[0049] Embodiment 2-4, preparation of glucose dehydrogenase-copper phosphate crystal composite microspheres, cellobiose dehydrogenase-copper phosphate crystal composite microspheres and uricase-copper phosphate crystal composite microspheres

[0050]The Candida antarctica lipase B (CALB) in the preparation step 1 of Example 1 was replaced with glucose dehydrogenase, cellobiose dehydrogenase and uricase respectively, and other operations were the same.

[0051] The scanning electron microscope image of glucose dehydrogenase-copper phosphate crystal composite microspheres is as follows image 3 shown by image 3 It can be seen that the particle size of the composite microsphere is about 10 μm. The enzyme load in the glucose dehydrogenase-copper phosphate crystal composite microsphere is 5%.

[0052] Scanning electron microscope image of cellobiose dehydrogenase-copper phosphate crystal composite microspheres Figure 4 shown by Figure 4 It can be seen that the particle size ...

Embodiment 5

[0054] Embodiment 5, glucose oxidase (GO X )-horseradish peroxidase (HRP)-copper phosphate crystal composite microspheres

[0055] 1. Prepare the PBS buffer solution (pH=7.4) of glucose oxidase (GOx) and horseradish peroxidase (HRP) with a total concentration of 0.5 mg / mL respectively (GO X and HRP concentrations are 0.25mg / mL) and 200mM copper sulfate pentahydrate aqueous solution.

[0056] 2. Take 5mL of the enzyme solution in step 1 into a glass vial, add 0.05mL of the copper sulfate solution in step 1, mix well, and place it in a 25°C incubator for 24h.

[0057] 3. Collect the precipitate after filtering the mixture obtained in step 2, wash with deionized water 3 times, filter again to collect the precipitate, freeze-dry for 5 hours to obtain glucose oxidase (GOx)-horseradish peroxidase (HRP)-copper phosphate Crystalline composite microspheres.

[0058] The scanning electron microscope picture of the composite microsphere prepared in this embodiment is as follows Figu...

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Abstract

The invention discloses an enzyme-inorganic crystal compound microsphere and a preparation method thereof. The preparation method of the enzyme-inorganic crystal compound microsphere provided by the invention comprises the following step: dissolving metal ions and the enzyme in a phosphate buffer to obtain the enzyme-inorganic crystal compound microsphere, wherein the metal ions are any one of calcium ions, magnesium ions, iron ions, ferrous ions, cupric ions, nickelous ions or zinc ions, and the phosphate buffer comprises phosphate dihydrogen ions or dihydrogen phosphate ions. The preparation method of the enzyme-inorganic crystal compound microsphere provided by the invention has the advantages of simplicity in operation, low cost and the like. The stability of the enzyme in the enzyme-inorganic crystal compound microsphere provided by the invention is improved and the activity of the enzyme is substantially maintained unchanged or even improved.

Description

technical field [0001] The invention relates to an enzyme-inorganic crystal composite microsphere and a preparation method thereof, belonging to the technical field of immobilized enzymes. Background technique [0002] As a kind of biocatalyst, enzyme has been widely used in the fields of medicine, food processing and fine chemical synthesis because of its high selectivity and mild reaction conditions. The multienzyme is composed of two or more different enzymes, which can carry out continuous catalytic reactions. The multi-enzyme system exhibits high efficiency and coordination in the catalytic process. However, the single or multiple enzymes used in industrial catalysis show defects such as low stability, poor tolerance, and non-recyclability under actual industrial operating conditions, which largely limit their industrial applications. Immobilized enzyme technology provides a way to solve the above problems. [0003] At present, immobilized enzyme technology has been ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
Inventor 李致贤戈钧刘铮
Owner TSINGHUA UNIV
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