Method for preparing immobilized enzyme by using spherical bacterial cellulose as carrier

A bacterial cellulose, immobilized enzyme technology, applied in microorganism-based methods, biochemical equipment and methods, immobilized on/in organic carriers, etc., can solve the problems of bacterial cellulose pellet immobilization enzymes, etc., Achieve the effect of good biodegradability, strong controllability and simple method

Inactive Publication Date: 2010-05-12
DONGHUA UNIV
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Problems solved by technology

At present, the immobilization carriers of enzymes mainly include inorganic carrier materials (such as silica, activated carbon, porous glass, etc.), organic synthetic polymer materials (such as polyacrylamide, polyethylene, ...

Method used

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  • Method for preparing immobilized enzyme by using spherical bacterial cellulose as carrier
  • Method for preparing immobilized enzyme by using spherical bacterial cellulose as carrier
  • Method for preparing immobilized enzyme by using spherical bacterial cellulose as carrier

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Embodiment 1

[0031] Pick 1 loop of single bacterium colony to inoculate in 100mL aseptic seed culture medium from the slope of Acetobacterium aceticum, described seed culture medium comprises (mass percentage): glucose 2.0%, yeast powder 0.5%, tryptone 0.5%, Na 2 HPO 4 12H 2 O0.27%, 0.115% citric acid, pH5.0, cultured at 30°C, 130rpm for 12h. The seed solution was inserted into a 500mL Erlenmeyer flask containing 300mL sterile fermentation medium with a 6% inoculum amount. Described fermentation medium comprises (mass percentage): glucose 2.0%, yeast powder 0.5%, tryptone 0.3%, Na 2 HPO 4 12H 2O 0.27%, citric acid 0.115%, pH 5.0; 30°C, 110rpm after culturing for 72 hours, many bacterial cellulose pellets were formed in the medium. After decanting the culture medium, take out the cellulose pellets, rinse them repeatedly with deionized water to remove the culture medium and impurities on the surface of the pellets, then soak the pellets in 0.1% NaOH solution, and treat them at 80°C for ...

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Abstract

The invention relates to a method for preparing immobilized enzyme by using spherical bacterial cellulose as a carrier. The method comprises the following steps of: inoculating activated slant strains into a seed culture medium for cultivation to obtain a liquid seed, shifting the liquid seed into a fermentation medium under a condition of 80 to 200 rpm at the temperature between 25 and 30 DEG C for 48 to 96 hours, and carrying out decantation to obtain bacterial cellulose balls; processing the bacterial cellulose balls in a water bath at the temperature between 80 and 90 DEG C for 30 to 120 min with 0.1 percent NaOH, rinsing the balls with deionized water, collecting the balls, neutralizing residual alkali liquor in the balls with 0.1 percent acetic acid, standing, rinsing the balls with the deionized water, taking the balls out, absorbing surface moisture, and freezing and drying the balls to obtain cellulose balls; and finally, preparing the immobilized enzyme by a physical adsorption method or an adsorption-crosslinking method. The spherical bacterial cellulose balls can effectively adsorb biological enzyme, maximally keep enzyme activity, and have safety and environmental protection. The method is simple and easy to operate and has strong controllability.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzymes, in particular to a method for preparing immobilized enzymes with spherical bacterial cellulose as a carrier. Background technique [0002] Because the enzyme has better substrate specificity, it has important application value in many fields. However, the free enzyme protein is susceptible to denaturation and inactivation under the influence of environmental conditions in the actual application process, and it is not easy to separate from the reaction system to achieve the purpose of repeated use, which limits the industrial application of enzymes to a certain extent. Enzyme immobilization technology is an effective means to achieve repeated and continuous use of enzymes and improve their stability. At present, the immobilization carriers of enzymes mainly include inorganic carrier materials (such as silica, activated carbon, porous glass, etc.), organic synthetic polymer materials (...

Claims

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Application Information

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IPC IPC(8): C12N11/12C12P19/04C12R1/41C12R1/38C12R1/01C12R1/02
Inventor 洪枫邹敏杨光杨雪霞曹张军杜明
Owner DONGHUA UNIV
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