Method for preparing immobilized nitrilase

A technology of nitrilase and immobilized nitrile, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of large material transfer resistance, unfavorable sustainable development, low enzyme reaction speed, etc., and achieve low production cost, Conducive to sustainable development and high resource utilization

Inactive Publication Date: 2012-06-27
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main disadvantages of this method are: cells are used as immobilized carriers, and immobilized nitrilase is distributed in the cells. Since the dead cells do not have well-developed pores, the material transfer resistance is large and the enzyme reaction speed is low; the rigid

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  • Method for preparing immobilized nitrilase

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Effect test

Embodiment 1

[0022] A method for preparing immobilized nitrilase, the specific method steps are as follows:

[0023] (1) Pretreatment of nitrilase

[0024] Using commercially available nitrilase as raw material, according to the mass (g) of commercially available nitrilase: the volume (mL) ratio of buffer solution is 1:10, add commercially available nitrilase into the buffer solution, stir to dissolve After 40 minutes, it is pumped into the suction filter for suction filtration, and the suction filtrate and the suction filter residue are collected respectively. The collected suction filtrate is used to prepare immobilized nitrilase for treatment; the collected suction filter residue is dried and used to add animal feed. Wherein, the buffer solution is a borax-calcium chloride buffer solution with a pH value of 8.0.

[0025] (2) Preparation of immobilized nitrilase crude product

[0026] After the (1) step was completed, according to the quality (g) of the commercially available lysine-m...

Embodiment 2

[0032] A method for preparing immobilized nitrilase, with embodiment 1, wherein:

[0033] In the 1st step, the mass of commercially available nitrilase: the volume ratio of buffer solution is 1g: 9mL, stirring and dissolving for 35 minutes, wherein, described buffer solution is the borax-potassium chloride buffer solution with a pH value of 9.0 .

[0034] In (2) step, the quality of commercially available lysine-macroporous ion exchange resin carrier: the volume of the suction filtrate collected in (1) step: glutaraldehyde mass concentration is the volume ratio of 25% glutaraldehyde solution 1g: 2.5mL: 0.7mL, soak in buffer solution for 2.5 hours, heat up to 35°C, and react at constant temperature for 6.5 hours, wherein the buffer solution is a borax-potassium chloride buffer solution with a pH value of 9.0.

[0035] In step (3), the quality of the suction filter residue collected in step (2): the volume ratio of distilled water is 1g: 4.5mL, and the collected washing residue...

Embodiment 3

[0038] A method for preparing immobilized nitrilase, with embodiment 1, wherein:

[0039]In the 1st step, the quality of commercially available nitrilase: the volume ratio of buffer solution is 1g: 8mL, stirring and dissolving 30 minutes, wherein, described buffer solution is the borax-calcium carbonate buffer solution that pH value is 10.8.

[0040] In (2) step, the quality of commercially available lysine-macroporous ion exchange resin carrier: the volume of the suction filtrate collected in (1) step: glutaraldehyde mass concentration is the volume ratio of 25% glutaraldehyde solution 1g: 3mL: 0.8mL, soak in the buffer solution for 3 hours, heat up to 45°C, and react at constant temperature for 7 hours, wherein the buffer solution is a borax-calcium carbonate buffer solution with a pH value of 10.8.

[0041] In step (3), the quality of the suction filter residue collected in step (2): the volume ratio of distilled water is 1g: 5mL, and the collected washing residue is stored...

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Abstract

The invention which relates to a method for preparing an immobilized nitrilase belongs to the technical field of immobilized enzymes. According to the invention, the immobilized nitrilase is prepared by treating a nitrilase sold in the market as a raw material and a lysine-macroporous ion exchange resin sold in the market as a carrier, and carrying out simple processes of immobilization, suction filtration and lavation. The method of the invention has the advantages of simplicity and easy implementation, mild reaction condition, simple and easily available equipment, no discharge of "three wastes", full substance resource utilization, low production cost, and convenient popularization and application; and the immobilized nitrilase has the advantages of small steric hindrance effect, high heat stability, wide use range, and long service life. The method can be widely used for preparing the immobilized enzymes, and products prepared through adopting the method of the invention can be widely applied to the biodegradation industry, the bioremediation industry, the pharmaceutical industry, the chemical engineering industry, the agricultural industry and the like.

Description

1. Technical field [0001] The invention belongs to the technical field of immobilized enzymes, and in particular relates to a method for preparing immobilized nitrilase with a commercially available lysine-macroporous ion exchange resin as a carrier. 2. Background technology [0002] Immobilized enzyme is a new type of biocatalyst, which is an enzyme derivative with enzyme catalytic properties bound to a water-insoluble carrier after water-soluble enzyme is treated by physical or chemical methods. Compared with free enzymes that can only be used once, immobilized enzymes not only have the catalytic properties of free enzymes, but also have the advantages of good stability and reusability, and can also realize continuous and automatic production processes. Direction of development. Nitrilase is a biocatalyst with a wide range of substrate adaptability, which can catalyze the hydrolysis of nitriles to generate corresponding carboxylic acids, realize chemical, regio and stereo...

Claims

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Application Information

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IPC IPC(8): C12N11/08
Inventor 周小华张杰康宏宽王丹徐溢王颖利
Owner CHONGQING UNIV
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