Method for immobilizing lipase using microstructure in hydrophilic/ hydrophobic composite membrane

A technology of immobilized lipase and composite membrane, applied in biochemical equipment and methods, immobilized on/in organic carriers, hydrolytic enzymes, etc. High cost and other issues, to achieve the effect of easy loading, high energy recovery rate and simple method

Inactive Publication Date: 2006-07-12
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chemical methods (cross-linking and covalent bonding) to immobilize enzymes will inevitably lead to the loss of some active sites of the enzyme, thereby reducing the activity of the enzyme.
At the same time, it also brings about the high cost of preparation, and the problem that the carrier cannot be reused
Although the adsorption method can keep the activity of the enzyme very well, due to the limitation of the adsorption performance of the material, the enzyme loading is too small, so that the activity of the immobilized enzyme membrane per unit area is low.

Method used

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  • Method for immobilizing lipase using microstructure in hydrophilic/ hydrophobic composite membrane
  • Method for immobilizing lipase using microstructure in hydrophilic/ hydrophobic composite membrane
  • Method for immobilizing lipase using microstructure in hydrophilic/ hydrophobic composite membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] A polytetrafluoroethylene membrane with an average pore diameter of 0.1 μm, a porosity of 50%, and a thickness of 85 μm was used as the hydrophobic layer. The mass fraction of the film-making solution for the hydrophilic layer is 15%, and the thickness of the cellulose acetate hydrophilic layer is controlled to 65 μm. It is volatilized at a temperature of 25°C and a humidity of 40% for 20 minutes, and then immersed in a deionized hydrogel bath to solidify and form a film. The lipase was dissolved in 0.05M sodium phosphate buffer solution with pH 7.5 to obtain a lipase solution with an enzyme protein concentration of 167mg / L. Set the area to 26cm 2 The membrane is loaded into a filter, 15ml of lipase solution is added, and the filter is carried out at 0.6MPa. The composite membrane has a primary filtration rejection rate of lipase of 20%, and the lipase protein loading is 0.0201mg / cm 2 immobilized lipase membrane.

[0020] The immobilized enzyme membrane is packed int...

Embodiment 2

[0021] Example 2: A polytetrafluoroethylene membrane with an average pore diameter of 0.1 μm, a porosity of 50%, and a thickness of 85 μm was used as the hydrophobic layer. The mass fraction of the film-making solution for the hydrophilic layer is 15%, and the thickness of the cellulose acetate hydrophilic layer is controlled to 135 μm. It is volatilized at a temperature of 25°C and a humidity of 40% for 20 minutes, and then immersed in a deionized hydrogel bath to solidify and form a film. The lipase was dissolved in 0.05M sodium phosphate buffer solution with pH 7.5 to obtain a lipase solution with an enzyme protein concentration of 167mg / L. Set the area to 26cm 2 The membrane is loaded into a filter, 15ml of lipase solution is added, and the filter is carried out at 0.6MPa. The primary filtration rejection rate of the composite membrane to lipase is 22%, and the lipase protein load is 0.0212mg / cm 2 immobilized lipase membrane.

[0022] The immobilized enzyme membrane is ...

Embodiment 3

[0024] A polytetrafluoroethylene membrane with an average pore diameter of 0.1 μm, a porosity of 50%, and a thickness of 85 μm was used as the hydrophobic layer. The mass fraction of the film-making solution for the hydrophilic layer is 10%, and the thickness of the cellulose acetate hydrophilic layer is controlled to 65 μm. It is volatilized at a temperature of 25°C and a humidity of 40% for 20 minutes, and then immersed in a deionized hydrogel bath to solidify and form a film. The lipase was dissolved in 0.05 M sodium phosphate buffer solution with pH 7.5 to obtain a lipase solution with an enzyme protein concentration of 84 mg / L. Set the area to 26cm 2 The membrane is put into a filter, 10ml of lipase solution is added, and the filter is performed at 0.3MPa. The primary filtration rejection of lipase by the composite membrane is 14.3%, and the lipase protein loading is 0.0046mg / cm 2 immobilized lipase membrane.

[0025] Put the immobilized enzyme membrane into a two-phas...

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Abstract

The invention relates to a method of microstructure fixation lipase in the filed of enzyme fixation of biology projection. It dissolves hydrophilic film material acetate fiber into dimethyl form amide to prepare for hydrophilic film solution, it lays water repellency politef film on the glass plate, it spread coats hydrophilic film solution on its surface and uses dry-wet changing method to obtain the compensate film which is formed by fine and close hydrophilic layer and porous bleed layer, it uses filtering method and microstructure to achieve enzyme fixation.

Description

technical field [0001] The invention relates to a method for immobilizing lipase by utilizing the microstructure in a hydrophilic / hydrophobic composite membrane, and belongs to the technical field of enzyme immobilization in bioengineering. Background technique [0002] Lipase can catalyze various organic reactions such as ester hydrolysis or alcoholysis, ester synthesis, transesterification, polypeptide synthesis and polymer synthesis on the oil-water interface, and has been widely used in food, fine chemical and pharmaceutical industries. As an important biocatalyst, the effectiveness and economy of lipase applications largely depend on the immobilization of the enzyme. There are many carriers for immobilizing enzymes. The membrane is used as the carrier for lipase immobilization, which combines the separation function, carrier function and separation function of the membrane, and has incomparable advantages with other carriers. [0003] In the current study, various flat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N11/04C12N9/20
Inventor 王玉军徐坚骆广生戴猷元
Owner TSINGHUA UNIV
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