Method for preparing immobilized cellulase

A technology of cellulase and cellulose acetate, which is applied in the direction of fixing on/in the organic carrier, and can solve the problems of poor stability, easy inactivation, denaturation, and easy inactivation of free cellulase

Inactive Publication Date: 2015-05-13
SOYBEAN TECH DEV RES CENT HEILONGJIANG PROV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention aims at the miscibility of free enzymes in the reaction system, which is not only difficult to separate and recover, but also the enzymes in the free state are less stable to heat, strong acid, strong alkali, high ionic strength and organic solvents, etc. The production of cellulase is generally difficult, and it is difficult to use repeatedly or continuously. The present invention aims to solve the problems of easy inactivation and denaturation of free cellulase during the enzymatic degumming process, so the cellulase is immobilized

Method used

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specific Embodiment approach 1

[0008] Embodiment 1: The method for immobilizing cellulase is realized through the following steps: 1. Preparation of composite membrane: 5 g of cellulose acetate is dissolved in 100 mL of acetone. The prepared polypropylene membrane was soaked in acetone solution dissolved in cellulose acetate to form a film naturally, and the cellulose acetate / polypropylene composite membrane was rinsed with deionized water and dried several times for later use; 2. Enzyme immobilization: the enzyme solution Cellulase with a concentration of 0.5-2.5% and disodium hydrogen phosphate-potassium dihydrogen phosphate buffer are mixed at a pH of 2 to 6, and a cellulose acetate / polypropylene composite film is added. After stirring for 1~5h, add 0.35~0.55% glutaraldehyde for cross-linking reaction for 1~5h, take out the enzyme film and wrap it on the surface of the cylinder to make an immobilized cellulase roll; Activity: The activity of cellulase is 5.103 U / cm 2 , the immobilized cellulase roller c...

specific Embodiment approach 2

[0009] Embodiment 2: The difference between this embodiment and Embodiment 1 is that in step 2, the concentration of glutaraldehyde is selected to be 0.45-0.55%. Other steps are the same as in the first embodiment.

specific Embodiment approach 3

[0010] Embodiment 3: The difference between this embodiment and Embodiment 1 is that in step 2, the concentration of glutaraldehyde is selected to be 0.45-0.55%, and the immobilization temperature is 40-50°C. Other steps are the same as in the first embodiment.

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Abstract

The invention relates to a method for preparing immobilized cellulase. As free enzyme is mixed and dissolved in a reaction system and is difficult to not only separate and recover but also use repeatedly or continuously, free cellulase is immobilized. Cellulose acetate is immobilized on polypropylene composite films with different pore diameters, an effect of immobilizing the cellulose acetate to the composite films is observed through a scanning electron microscope, a polypropylene composite film with an optimal pore diameter is selected out, and the selected optimal composite film is immobilized on a roll. Conditions for immobilizing the free cellulase are that the glutaraldehyde concentration is selected to be 0.45-0.55%, the immobilization temperature is 48-49 DEG C, the pH (potential of hydrogen) value for immobilization is selected to be 3.9-4.0, the crosslinking time is 3-4 hours, the adsorption time is 3.0-3.1 hours, and the cellulase concentration is 1.0-2.0%. The activity of the free cellulase is measured to be 5.103 U / cm<2>, the enzyme activity still keeps about 80% after the roll for immobilizing the cellulase continuously presses soybean embryo slices for 24 hours, and the activity of the obtained immobilized cellulase is 4.732 U / cm<2>.

Description

technical field [0001] The invention relates to a method for preparing immobilized cellulase. Background technique [0002] Cellulase is an important enzyme product, a compound enzyme, mainly composed of exo-β-glucanase, endo-β-glucanase and β-glucosidase, etc., which has a high Xylanase activity. Due to the huge market potential of cellulase in the fields of feed, alcohol, textile and food, it has been favored by people in the industry at home and abroad. It will be the fourth largest industrial enzyme after glucoamylase, amylase and protease, even in China. It is entirely possible to become the largest enzyme species, so cellulase is a new growth point in the enzyme preparation industry. [0003] Enzyme immobilization is a kind of technology that binds or confines enzymes in a certain area with solid materials, and can still carry out its unique catalytic reactions, and can be recycled and reused. Compared with free enzymes, immobilized enzymes overcome the shortcomings...

Claims

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Application Information

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IPC IPC(8): C12N11/12C12N11/08
Inventor 孙树坤陈昊解桂东胡立志于殿宇王喜泉许慧郑环宇朱秀清姚磊
Owner SOYBEAN TECH DEV RES CENT HEILONGJIANG PROV
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