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Preparation method of attapulgite immobilized enzyme for ester exchange reaction

The technology of transesterification reaction and attapulgite is applied in the field of catalytic material and enzyme immobilization, which can solve the problems of lipase falling off gradually, immobilized enzyme activity decreasing, and the preparation process is not environmentally friendly enough, so as to achieve good performance in repeated use and high reaction efficiency. The effect of fewer by-products, avoiding the use of toxic solvents and corrosive catalysts

Active Publication Date: 2012-03-21
惠州市康维健生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports on using attapulgite as the carrier of immobilized enzymes. Patent 200610039833.4 discloses the adsorption and immobilization of lipase on the surface of attapulgite. Because only a single physical adsorption is used, lipase tends to fall off gradually during use. , resulting in a decrease in the activity of the immobilized enzyme
Shan Liang et al. used aminopropylsilane-modified attapulgite combined with glutaraldehyde to immobilize bromelain. However, the traditional silanization method requires the use of a large amount of toxic organic solvents such as toluene, and the reaction temperature is relatively high. The preparation process is not environmentally friendly
In addition, it has been reported in the literature that hydrophobic carriers are beneficial to improve the catalytic activity of immobilized lipase transesterification. However, pure hydrophobic materials (such as octyl silica gel) cannot be wetted in aqueous solution, and there will be operational difficulties during the immobilization process and the immobilization of the enzyme is not easy. uniform phenomenon
Sol-gel method is used to modify the surface of attapulgite to make it have dual retention of hydrophobicity and ion exchange, which can not only improve the adsorption capacity and stability of lipase, but also improve its catalytic activity for transesterification. Research in this area has not been reported

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A preparation method for attapulgite-immobilized enzymes for transesterification, comprising the steps of:

[0031] 1) Acid treatment of attapulgite (attapulgite is first acid-treated to remove metal impurities and activate surface hydroxyl groups): 50g attapulgite is added to 500mL hydrochloric acid solution, the concentration of hydrochloric acid solution is 4-6mol / L, soaking time 10h (soaking temperature 25°C), after the treatment, wash with pure water until neutral to obtain acid-treated attapulgite. The particle size of the attapulgite is 100 mesh.

[0032] 2) Surface chemical modification of attapulgite: suspend 10 g of acid-treated attapulgite in 500 mL of isopropanol to obtain a suspension with a concentration of 20 mg / mL;

[0033] The total amount of silylating agent added is 15 mmol, and the silylating agent is composed of isobutyltrimethoxysilane and mercaptopropyltrimethoxysilane, wherein the mole of isobutyltriethoxysilane and mercaptopropyltrimethoxysilan...

Embodiment 2

[0041] A preparation method for attapulgite-immobilized enzymes for transesterification, comprising the steps of:

[0042] 1) The acid treatment steps of the attapulgite are the same as in Example 1, and the particle size of the attapulgite is 200 mesh.

[0043] 2) Surface chemical modification of attapulgite: 30 g of acid-treated attapulgite was suspended in 750 mL of ethanol to obtain a suspension with a concentration of 40 mg / mL;

[0044] The total amount of silylating agent added is 20mmol, and the silylating agent is composed of octyltriethoxysilane and mercaptopropyltriethoxysilane, wherein the moles of octyltriethoxysilane and mercaptopropyltriethoxysilane The ratio is 1:1, add 20mL, 25%-28% concentrated ammonia water as a catalyst; mix the silylating agent and catalyst with the above suspension to obtain a mixture; react the mixture at 50°C for 15h under nitrogen protection, and the mechanical stirring speed is 300rpm ; After the reaction, the granular matter was sepa...

Embodiment 3

[0050] A preparation method for attapulgite-immobilized enzymes for transesterification, comprising the steps of:

[0051] 1) The acid treatment steps of the attapulgite are the same as in Example 1, and the particle size of the attapulgite is 300 mesh.

[0052] 2) Surface chemical modification of attapulgite: Suspend 25 g of acid-treated attapulgite in 500 mL of ethanol to obtain a suspension with a concentration of 50 mg / mL;

[0053] The total amount of silylating agent added is 25 mmol, and the silylating agent is composed of isobutyltrimethoxysilane and aminopropyltrimethoxysilane, wherein the molar ratio of isobutyltrimethoxysilane to aminopropyltrimethoxysilane 1:1.5, add 30mL, 25%-28% ammonia water as a catalyst; mix the silylating agent and catalyst with the above suspension to obtain a mixture; react the mixture at 60°C for 20h under nitrogen protection, and the mechanical stirring speed is 400rpm; After the end, the granular matter is separated by centrifugation, th...

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Abstract

The invention relates to a preparation method of attapulgite immobilized enzyme for ester exchange reaction. The preparation method of the attapulgite immobilized enzyme for the ester exchange reaction is characterized by comprising the following steps of: 1) performing acid treatment on attapulgite to obtain acid treated attapulgite; 2) performing surface chemical modification on the attapulgiteto obtain attapulgite granules with hydrophobic capacity and ion exchange double-adsorption capacity; and 3) immobilizing lipase on the surface of the attapulgite, namely putting the chemically modified attapulgite granules into a lipase solution in a ratio of (50-80)mg: 1mL, performing mechanical stirring and immobilization of the lipase, cleaning the reaction product by using distilled water after the reaction is finished, soaking the product into an organic solvent, performing centrifugal separation and performing freeze drying, and thus obtaining the attapulgite immobilized enzyme for theester exchange reaction. The attapulgite immobilized enzyme has the characteristics of low cost, high catalytic activity, good reuse performance and environment friendliness in the preparation process.

Description

technical field [0001] The present invention relates to a preparation method of attapulgite immobilized enzyme for transesterification reaction, in particular to a method for immobilizing enzyme by attapulgite modified by sol-gel method, which is used to improve the activity of enzyme The invention relates to immobilization stability and catalytic activity, belonging to the technical field of catalytic materials and enzyme immobilization. Background technique [0002] Grease transesterification refers to the chemical reaction of grease (including various natural vegetable oils, animal fats, and waste oil from the food industry) with low-carbon alcohols, fatty acids or / and fatty acid esters, phytosterols, etc. under the action of a catalyst. One of the important means of modification of oil and fatty acid derivatives, it can prepare a series of high value-added chemical products or health care products, such as fatty acid low-carbon alkyl esters (biodiesel), sterol esters, su...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
Inventor 黄凤洪郑明明郭萍梅李文林邓乾春杨湄黄庆德刘昌盛万楚筠钮琰星
Owner 惠州市康维健生物科技有限公司
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