Preparation method of immobilized cellulase

A cellulase and weighing technology, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of easy detachment of the enzyme molecule from the carrier, weak binding force between the enzyme and the carrier, and poor stability, and achieve The effect of high immobilization rate, good stability and good mechanical strength

Inactive Publication Date: 2012-05-16
王春杰
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a kind of fiber to solve the defects of poor mechanical strength, poor stability, weak binding force between the enzyme and the carrier, and the enzyme molecule is easy to fall off from the carrier during the use process. Sulfase immobilization method

Method used

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specific Embodiment approach 1

[0008] Specific embodiment one: the preparation method of the immobilized cellulase of this embodiment is realized according to the following steps: one, take by weighing 2.0-4.0g deacetylation degree and be the chitosan of 90%, 0.6-0.8g PEG6000 is dissolved in 100ml2% acetic acid solution at 35-45°C for 1.5-2.5 hours; 2. Inject the blended solution into 2% sodium tripolyphosphate solution with a syringe and let it rest for 2.5-3.5 hours; 3. Filter and wash to obtain neutral spherical Chitosan-polyethylene glycol carrier; 4. Weigh 9-11g carrier, add 100ml 5% glutaraldehyde solution, activate at 45-55°C for 5.5-6.5h; 5. Wash with acetone and distilled water until neutral after filtration 6. Weigh 1.5-2.5g of the activated carrier, add 20ml of cellulose cosmetic solution and 17-19℃ for immobilization reaction for 15-17h; 7. Rinse the immobilized enzyme several times after filtering to obtain the immobilized cellulose enzyme.

[0009] In this embodiment, chitosan, sodium tripoly...

specific Embodiment approach 2

[0010] Specific embodiment two: the difference between this embodiment and specific embodiment one is that in step one, it is 90% chitosan to accurately take by weighing 3.0g degree of deacetylation, 0.75g PEG6000 is dissolved in the acetic acid solution of 100ml 2%, at 40 ℃ Blend for 2h. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0011] Embodiment 3: The difference between this embodiment and Embodiment 1 is that 2% sodium tripolyphosphate solution with pH=8.2 is used in step 2. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention discloses a preparation method of immobilized cellulase, relating to an enzyme immobilization method. The method comprises the following steps of: firstly, mixing chitosan with the degree of deacetylation of 90% with an acetic acid solution, then injecting the mixed solution into a 2% sodium tripolyphosphate solution, filtering and washing to obtain neutral spherical chitosan-polyethylene glycol carriers, then activating the carriers by a glutaraldehyde solution, carrying out immobilization reaction of the carriers and cellulase, and filtering and washing to obtain immobilized cellulase. The immobilized cellulase prepared by the method has the advantages of high mechanical strength, good enzyme stability, high enzyme activity, strong binding force between the enzyme and the carriers and high rate of immobilization.

Description

technical field [0001] The invention relates to a biocatalyst immobilization method, more precisely a cellulase immobilization method, which belongs to the technical field of biological reagent preparation. Background technique [0002] Natural cellulose is the most abundant renewable organic matter on the earth. The plant matter produced through photosynthesis in the world is about 1 trillion tons per year, 89% of which are not currently used by humans. From the perspective of human utilization of natural resources, this Undoubtedly a huge waste. With the rapid growth of the world's population and the unsolved food and energy crisis, the utilization of cellulose resources has aroused great concern and great attention from all countries in the world. [0003] Ordinary organisms cannot directly use cellulose materials as food, and can only be used by hydrolyzing cellulose into glucose. The general methods for hydrolyzing cellulose into glucose include acid hydrolysis and en...

Claims

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Application Information

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IPC IPC(8): C12N11/10
Inventor 王春杰
Owner 王春杰
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