Penioillin acylase preparation and immobilization method

A penicillin acylase and preparation technology, applied in the field of materials, can solve the problems of low recovery rate and loss of enzyme activity of immobilized enzymes, and achieve the effects of easy recycling, high loading rate, and easy availability

Active Publication Date: 2018-03-02
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
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Problems solved by technology

However, glutaraldehyde has a great influence on the activity of the enzyme, and the recovery rate of the immobilized enzyme enzyme activity obtained by glutaraldehyde crosslinking is low, and the C=N bond formed by glutaraldehyde and the amino group on the surface of the carrier is easily hydrolyzed, especially Even when the temperature is high, the enzyme activity will be seriously lost during the immobilization process.

Method used

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  • Penioillin acylase preparation and immobilization method
  • Penioillin acylase preparation and immobilization method
  • Penioillin acylase preparation and immobilization method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) getting quality is that 2.00g tetrakis hydroxymethyl phosphorus chloride is added in 90ml water, obtains tetrakis hydroxymethyl phosphorus chloride aqueous solution; Getting quality is that 0.56g potassium hydroxide is added in 10ml water, obtains potassium hydroxide aqueous solution ;

[0038] (2) At room temperature, slowly drop an aqueous potassium hydroxide solution into an aqueous tetrakishydroxymethyl phosphorus chloride solution to obtain a mixed solution A;

[0039] (3) Add 2.00 g of magnetic cellulose nanocrystals to the mixed solution A, stir at room temperature (the stirring speed is 150 rpm) for 8 minutes, centrifuge, and wash with deionized water to obtain the treated magnetic cellulose nanocrystals;

[0040] (4) 200.00 mg of penicillin acylase was dissolved in 50 ml of phosphate buffer with a pH of 7.5 to obtain an enzyme solution;

[0041] (5) Mix the processed magnetic cellulose nanocrystals obtained in step (3) with the enzyme solution configured in ...

Embodiment 2

[0044] (1) Get a quality of 7.00g of tetrakis hydroxymethyl phosphorus chloride and join it in 90ml water to obtain a tetrakis hydroxymethyl phosphorus chloride aqueous solution; get a quality of 1.96g of potassium hydroxide and join it in 10ml water to obtain an aqueous solution of potassium hydroxide ;

[0045] (2) At room temperature, slowly drop an aqueous potassium hydroxide solution into an aqueous tetrakishydroxymethyl phosphorus chloride solution to obtain a mixed solution A;

[0046] (3) Add 2.00 g of magnetic cellulose nanocrystals to the mixed solution A, stir at room temperature (the stirring speed is 150 rpm) for 8 minutes, centrifuge, and wash with deionized water to obtain the treated magnetic cellulose nanocrystals;

[0047] (4) 200.00 mg of penicillin acylase was dissolved in 50 ml of phosphate buffer with a pH of 7.5 to obtain an enzyme solution;

[0048] (5) Mix the processed magnetic cellulose nanocrystals obtained in step (3) with the enzyme solution conf...

Embodiment 3

[0050] (1) getting quality is that 2.00g tetrakis hydroxymethyl phosphorus chloride is added in 90ml water, obtains tetrakis hydroxymethyl phosphorus chloride aqueous solution; Getting quality is that 0.56g potassium hydroxide is added in 10ml water, obtains potassium hydroxide aqueous solution ;

[0051] (2) At room temperature, an aqueous potassium hydroxide solution is slowly added dropwise to an aqueous tetrakishydroxymethyl phosphorus chloride solution to obtain a mixed solution A;

[0052] (3) Add 2.00 g of magnetic cellulose nanocrystals to mixed solution A, stir at room temperature (150 rpm) for 8 min, centrifuge, and wash with deionized water to obtain treated magnetic cellulose nanocrystals;

[0053] (4) 600.00 mg of penicillin acylase was dissolved in 50 ml of phosphate buffer solution with a pH of 7.5 to obtain an enzyme solution;

[0054] (5) Mix the processed magnetic cellulose nanocrystals obtained in step (3) with the enzyme solution configured in step (4), st...

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Abstract

The invention belongs to the technical field of a material, and discloses a penioillin acylase preparation and an immobilization method. The immobilization method comprises the following steps of (1)under the conditions of stirring at room temperature, dripping a potassium hydroxide water solution into a tetrakis hydroxymethyl phosphonium chloride solution to obtain a mixed solution containing trisphosphine oxide; (2) adding magnetic cellulose nanometer crystals into the mixed solution containing trisphosphine oxide in the step (1); performing stirring for 1 to 30min at room temperature; performing centrifugal washing to obtain the treated magnetic cellulose nanometer crystals; (3) diluting the penioillin acylase into a buffer solution with the pH being 6 to 11 to obtain an enzyme solution; (4) adding the treated magnetic cellulose nanometer crystals into the enzyme solution; performing curing at room temperature to obtain the immobilized penioillin acylase, i.e., the penioillin acylase preparation. The method is simple; the cost is low; the enzyme activity recovery rate of the prepared immobilized penioillin acylase preparation is high; the enzyme loading rate is high; meanwhile,the stability is high.

Description

technical field [0001] The invention belongs to the technical field of materials, and relates to a method for immobilizing penicillin acylase and an enzyme preparation. Background technique [0002] Cellulose nanocrystal (CNC) is a natural nano-material with excellent properties, which is a rod-shaped crystal with a length of 50-1000nm and a width of 5-30nm obtained by acid hydrolysis of cellulose raw materials under certain conditions. Cellulose nanocrystals are extremely stable in solution and difficult to separate. The magnetic cellulose nanocrystal prepared by introducing the magnetic ferroferric oxide into the cellulose nanocrystal can be rapidly separated from the liquid phase under the action of a magnetic field. [0003] Magnetic cellulose nanocrystals have the advantages of high crystallinity, high degree of polymerization, high hydrophilicity, high mechanical strength, high biocompatibility, high specific surface area, easy recycling, etc., and have great potentia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12
CPCC12N9/84C12N11/12C12Y305/01011
Inventor 宗敏华黄子轩娄文勇熊隽尹航
Owner SOUTH CHINA UNIV OF TECH
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