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Cell immobilization method of nitrilase producing strain

A kind of nitrilase, the technology of producing bacteria, which is applied in the application field of bioengineering technology

Inactive Publication Date: 2011-07-13
WUXI HENGYU BIOLOGICAL TECH
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  • Abstract
  • Description
  • Claims
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example 1

[0008] 1. Preparation of nitrilase-producing bacteria liquid

[0009] Liquid medium: potassium dihydrogen phosphate 1g, magnesium sulfate 0.1g, ammonium chloride 0.5g, ferrous sulfate 0.015g, manganese chloride 0.01g, cobalt chloride 0.01g, sodium chloride 0.5g, caprolactam 1.5g, yeast Cream 5g, peptone 5g, glycerin 5g, add water to 500mL, pH7.0;

[0010] Cultivation method: pick bacteria from the slant strains with an inoculation loop, inoculate them in the seed medium of a shaker flask, store 150mL of culture medium in a 500mL triangular flask at 30°C, 200r / min, and shake for 2 days;

[0011] 2. Cell Immobilization

[0012] Centrifuge the prepared bacterial solution to obtain the bacterial cells, wash with distilled water and then centrifuge, take 0.02 g of bacterial cells with a dry weight and suspend them in Tris-Hcl buffer solution with pH 7.0, and add 2.5 % and 5% sodium alginate and PEG400 blends, after mixing, use a syringe to inject a concentration of 0.1MCaCl 2 In...

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Abstract

The invention belongs to the field of biological engineering technology application, and in particular relates to a cell immobilization method of a nitrilase producing strain. Sodium alginate / polyethylene glycol (PEG) 400 beads are white, and have the diameter of 1 to 2mm. The cell immobilization method comprises the following steps of: culturing the nitrilase producing strain in liquid to obtain strain liquid, performing centrifugal separation to obtain thalli, suspending the thalli in triisopropylphenylsulfonyl (Tris)-Hcl buffer solution, adding a sterilized blend of alginate and PEG400, uniformly mixing, stably injecting into CaCl2 solution by using an injector, and immobilizing. The cell immobilization method provides a novel way for cell immobilization research and has a certain application value.

Description

technical field [0001] The invention relates to a method for cell immobilization of nitrilase-producing bacteria. It belongs to the application field of bioengineering technology. Background technique [0002] Nitrile compounds are a class of major pollutants, and the conversion of cyano groups into carboxyl groups by nitrile group hydrolysis cells can not only reduce environmental pollution, but also obtain carboxylic acid substances as a class of very useful compounds. Therefore, it is very attractive to choose suitable materials to obtain immobilized cells with high activity, good stability and repeated use. Compared with free cells, immobilized cells have lower activity. This is because there are greater or less mass transfer resistances depending on the materials used for immobilized cells. Overcoming these mass transfer resistances is the key to improving the activity of immobilized cells. point. Therefore, various methods are used to improve the permeability of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04
Inventor 陈敬华邓超马莉
Owner WUXI HENGYU BIOLOGICAL TECH
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