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Preparation method of immobilized lipase using clay as carrier

A technology of immobilizing lipase and lipase, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of being easily restricted by mass transfer resistance and easy leakage of enzymes, and achieves abundant reserves, mild conditions and wide distribution Effect

Inactive Publication Date: 2015-03-11
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Encapsulation enzymes are easy to leak, and the catalytic reaction process is easily limited by mass transfer resistance, which is only suitable for the reaction of small molecule substrates and products

Method used

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  • Preparation method of immobilized lipase using clay as carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: take clay as the preparation method of the immobilized lipase of carrier, the method is as follows:

[0034] (1) Pretreatment of clay carrier: mix 20g of clay with 12g of water, and gradually add 6g of polyacrylamide under constant speed agitator stirring. After mixing, the slurry is formed and dried in the shade for 48 hours. Calcined at 600°C for 3 hours, crushed, and sieved into particles with a particle size of 100 mesh;

[0035] (2) Hydroxylation treatment of clay carrier: Take 10g of pretreated clay, add 130ml of 1mol / L hydrochloric acid to it, and fully stir in a water bath at 75°C for 15min, then filter with suction, and wash the obtained clay with deionized water To the pH of the washing solution = 7, dry at 110°C for later use;

[0036] (3) Silanization of clay carrier: Take 10g of hydroxylated clay, add 30ml of petroleum ether and 0.3mL of methacryloxypropyl trimethoxysilane to it, mix well, and place it on a shaker at room temperature Rea...

Embodiment 2

[0039] Embodiment 2: take clay as the preparation method of the immobilized lipase of carrier, the method is as follows:

[0040] (1) Pretreatment of clay carrier: Mix 20g of clay with 16g of water, and gradually add 10g of polyacrylamide under stirring with a constant speed mixer. After mixing, the slurry is formed and dried in a cool place for 60 hours. Calcined at 700°C for 2 hours, crushed, and sieved into particles with a particle size of 100 mesh;

[0041] (2) Hydroxylation treatment of clay carrier: take 10g of pretreated clay, add 120ml of 2mol / L hydrochloric acid to it, and fully stir in a water bath at 80°C for 10min, then filter with suction, and wash the obtained clay with deionized water To the pH of the washing solution = 7, dry at 110°C for later use;

[0042] (3) Silanization of clay carrier: Take 10g of hydroxylated clay, add 30ml of petroleum ether and 0.4mL of 3-aminopropyltriethoxysilane to it, mix thoroughly, and place it in a shaker at room temperat...

Embodiment 3

[0045] Embodiment 3: the preparation method of the immobilized lipase with clay as carrier, the method is as follows:

[0046] (1) Pretreatment of clay carrier: mix 20g of clay with 12g of water, and gradually add 10g of polydimethyldiallylammonium chloride under constant speed agitator stirring. After mixing, the slurry is formed and placed in a cool place After drying for 48 hours, the dried clay was calcined at 600°C for 3 hours, crushed, and sieved into particles with a particle size of 100 mesh;

[0047] (2) Hydroxylation treatment of clay carrier: Hydroxylation treatment of clay carrier: Take 10g of pretreated clay, add 100ml of 3mol / L hydrochloric acid to it, stir fully in a water bath at 75°C for 15min, and filter with suction to obtain Wash the clay with deionized water until the pH of the washing solution is 7, and dry it at 110°C for later use;

[0048] (3) Silanization of clay carrier: Take 10g of hydroxylated clay, add 40ml of petroleum ether and 0.3mL of me...

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Abstract

The invention discloses a preparation method of immobilized lipase using clay as a carrier. The preparation method comprises the following steps of: firstly, uniformly mixing the clay with water and a flocculating agent, filling the obtain slurry into a mold to be molded, drying and calcining the clay at a certain temperature; subsequently crushing and screening; respectively conducting hydroxylation and silanization treatment on the screened clay; adding the treated clay as the carrier into a lipase solution which is prepared from a buffer solution; oscillating and adsorbing by using a shaker; after the adsorption is accomplished, washing the clay by using the buffer solution till no protein can be eluted; and freeze-drying so as to obtain the immobilized lipase. The clay adopted in the method has the advantages of easily-accessible raw material and low price, meanwhile has good physical adsorbability and surface chemical activity, and can remarkably improve the adsorptive property of the carrier when being subjected to modification treatment; and the prepared immobilized lipase is not only high in reaction enzyme activity, but also has better catalytic activity than free lipase.

Description

technical field [0001] The invention relates to the technical field of immobilized lipase, in particular to a preparation method of immobilized lipase with clay as a carrier. Background technique [0002] Lipase (Lipase, EC 3.1.1.3) is a type of triglyceride hydrolase, which can catalyze the hydrolysis of natural oils to generate fatty acids, glycerol, monoglycerides or diglycerides, and can complete hydrolysis, esterification, transesterification, Reactions such as transesterification and synthesis are widely used in oil hydrolysis, food processing, food quality improvement, disease diagnosis, catalytic drug synthesis, leather production, research and development of biodegradable non-polluting detergents, and biodiesel production. However, in actual use, free enzymes have disadvantages such as easy inactivation, unstable catalytic reaction, difficult separation from products, or high separation costs even under optimal reaction conditions. In addition, lipase enzyme prepar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/14
Inventor 余旭亚邓涛游金坤
Owner KUNMING UNIV OF SCI & TECH
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