Method for preparing magnetic cellulose immobilized lysozyme

A technology based on cellulose and cellulose solution is applied in the field of preparation of magnetic-based cellulose immobilized lysozyme, which can solve problems such as difficulty in recovering immobilized enzymes, improve dehydration effect, simple immobilization operation, and reduce weight reduction. cost effect

Inactive Publication Date: 2018-09-04
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The immobilized lysozyme lysed sludge system is a heterogeneous system, which makes it difficult to recover the immobilized enzyme
In a heterogeneous reaction system, there is no relevant report on the method of using magnetic cellulose microspheres to immobilize lysozyme to lyse sludge

Method used

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  • Method for preparing magnetic cellulose immobilized lysozyme
  • Method for preparing magnetic cellulose immobilized lysozyme
  • Method for preparing magnetic cellulose immobilized lysozyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039](1) Get 1.35g ferric chloride, 3.6g sodium acetate and 1.0g polyethylene glycol and join in 40ml of ethylene glycol and diethylene glycol mixed solvent (volume ratio is 2:1), mechanically stir, obtain clarification The yellow solution was transferred to a high-temperature reaction kettle made of polytetrafluoroethylene, sealed, and placed in a high-temperature drying oven at 200°C for 8 hours, cooled to room temperature, separated by an external magnetic field, washed several times with methanol and water, and frozen Dry to constant weight to obtain nano ferric oxide.

[0040] (2) Prepare a solution of 100g of sodium hydroxide, urea and water mixed at a mass ratio of 7:10:83, pre-cool to a low temperature of -8°C to -12°C, add 5g of microcrystalline cellulose, and stir rapidly to form a transparent and uniform Cellulose solution, then add nanometer iron ferric oxide in 0.3g step (1), stir evenly, remove air bubble, obtain ferric oxide-cellulose solution, add dropwise to ...

Embodiment 2

[0045] Take 6 parts of 1ml of the magnetic-based cellulose nanosphere solution in Example 1, add 400 μL of 1% glutaraldehyde (crosslinking agent) and 4.6mL of pH7.0 phosphate buffer solution, and vibrate on a shaker at 25°C for crosslinking respectively 1h, 2h, 4h, 6h, 8h, 10h, get immobilized carrier after magnetic separation and filtration, wash with phosphate buffer three times, add 100μL 1mg / mL lysozyme solution, shake and adsorb at 150~200r / min, 25℃ After 4 hours, apply an external magnetic field to separate, and wash three times with a phosphate buffer solution with a pH value of 7 to obtain magnetic-based cellulose-immobilized lysozyme. Determination of the immobilized lysozyme enzyme activity, such as Image 6 shown. It can be seen from the figure that when the crosslinking time is 4h, the activity of the immobilized lysozyme is the highest (12U / g), and the recovery rate of the activity of the immobilized lysozyme reaches the maximum (47.8%).

Embodiment 3

[0047] Take 8 parts of 1ml of the magnetic-based cellulose nanosphere solution in Example 1, add 400 μL of 1% glutaraldehyde (crosslinking agent) and 4.6mL of pH7.0 phosphate buffer solution, and vibrate on a shaker at 25°C for crosslinking respectively 4h. The immobilized carrier was obtained after magnetic separation and filtration, washed three times with phosphate buffer, added 100 μL, 200 μL, 400 μL, 500 μL, 600 μL, 700 μL, 800 μL, 1000 μL 1 mg / mL lysozyme solution (1 mg / mL), at 150 ~ 200 r / min at 25°C for 4 hours, separated by an external magnetic field, and washed three times with phosphate buffer with a pH value of 7 to obtain magnetic-based cellulose-immobilized lysozyme. Determination of the immobilized lysozyme enzyme activity, such as Figure 7 shown. It can be seen from the figure that when the enzyme dosage reaches 500 μL (1 mg / mL), the activity recovery rate reaches the highest, which is 68.5%.

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Abstract

The invention relates to a method for preparing magnetic cellulose immobilized lysozyme. The method comprises the following steps: preparing magnetic cellulose nanoparticles, preparing an immobilizedcarrier, and preparing the magnetic cellulose immobilized lysozyme. The immobilized operation is simple, the prepared magnetic cellulose immobilized lysozyme has the advantages of uniform shape, largespecific surface area, high enzyme activity recovery rate, and easy storage, can be used for lysis sludge reduction, also has the advantages of easy recovery, repeated usage and good stability, can reduce the lysis sludge reduction cost by a biological method, and overcomes the disadvantages of difficult enzyme recovery and high cost in a traditional biological method lysis technology.

Description

technical field [0001] The invention belongs to the field of lysozyme immobilization methods, in particular to a preparation method of magnetic-based cellulose immobilized lysozyme. Background technique [0002] At present, the activated sludge method is widely used in urban sewage treatment (sewage treatment plants), and a large amount of excess activated sludge (WAS) with a water content exceeding 95% is produced during the treatment process. Therefore, the dehydration treatment of sludge is the most direct and effective measure to reduce the amount of test sludge, and it is an indispensable link in the sludge treatment process. How to realize rapid dehydration and maximum dehydration of sludge has become a major issue of great concern to the environmental circles. [0003] Activated sludge is a complex aggregate, mainly composed of organic debris, bacterial cells, infinite particles, colloids and other components, which are embedded in a network structure composed of ext...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12C12N9/36C02F11/02
CPCC02F11/02C02F2303/06C12N9/2462C12N11/12C12Y302/01017
Inventor 李登新薛飞李玉龙郭庆峰阳光辉郭启浩孙红蕊
Owner DONGHUA UNIV
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