Method for improving activity and stability of immobilized lipase

A technology of immobilized lipase and immobilized enzyme, which is applied in the field of enzyme chemistry, can solve problems such as difficult separation, easy recovery of glycerin, and less alcohol consumption, and achieve simple operation, high promotion and utilization value, and improved catalytic activity and stability Effect

Inactive Publication Date: 2010-06-16
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using lipase as a catalyst to prepare biodiesel, the reaction conditions are mild, the amount of alcohol is less, glycerin is easy to recover, and there is no waste, which can solve the problems of difficult separation and high energy consumption of the catalysts used in the production of biodiesel by chemical methods

Method used

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  • Method for improving activity and stability of immobilized lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] a, lipase liquid preparation: get 15g enzyme powder and be dissolved in the phosphate buffer solution (0.05mol / LK) of 300ml, pH7.0 2 HPO 4 -KH 2 PO 4 ), to prepare the lipase liquid of 0.05mg / ml.

[0026] b. Immobilization of lipase: Add 1 g of pretreated 40 mesh activated carbon into 30 ml of enzyme solution, shake and adsorb at 30°C and 150 rpm for 2 hours. The pretreatment is soaking with 1mol / L HCl at room temperature for 3h, and then washing with water until neutral.

[0027] c. Separation and storage of immobilized lipase: the wet immobilized enzyme was separated by filtration and placed at 4°C for 24 hours.

[0028] d. Organic solvent treatment: choose acetone as the immobilized enzyme treatment agent. Add 1 g of wet immobilized enzyme to 30 ml of acetone, shake and treat at 25°C and 150 rpm for 1 hour, filter the treated immobilized enzyme, place it at 4°C for 24 hours, and measure the enzyme activity.

[0029] e. Soybean oil transesterification: Mix 22.5g...

Embodiment 2

[0033] a, lipase liquid preparation: get 15g enzyme powder and be dissolved in the phosphate buffer solution (0.05mol / LK) of 300ml, pH7.0 2 HPO 4 -KH 2 PO 4 ), to prepare the lipase liquid of 0.05mg / ml.

[0034] b. Immobilization of lipase: Add 1 g of pretreated silica gel G into 40 ml of enzyme solution, shake and adsorb at 30° C. and 150 rpm for 2 h. The silica gel G pretreatment method is the same as that in Example 1.

[0035] c. Separation and storage of immobilized lipase: the wet immobilized enzyme was separated by filtration and placed at 4°C for 24 hours.

[0036] d. Use ethyl acetate as the immobilized enzyme treatment agent, add 1g of wet immobilized enzyme to 20ml ethyl acetate, shake at 30°C and 150rpm for 1.5h, filter the treated immobilized enzyme, and place it at 4°C 24h, measure enzyme activity.

[0037]e. Soybean oil transesterification: Mix 22.5g soybean oil, 11ml n-hexane and 1g organic solvent-treated immobilized enzyme, preheat at 30°C for 20min, an...

Embodiment 3

[0041] a, lipase liquid preparation: get 15g enzyme powder and be dissolved in the phosphate buffer solution (0.05mol / LK) of 300ml, pH7.0 2 HPO 4 -KH 2 PO 4 ), to obtain a lipase solution of 0.05 mg / ml.

[0042] b. Immobilization of lipase: Add 1 g of pretreated DM-130 resin into 50 ml of enzyme solution, shake and adsorb at 30° C. and 150 rpm for 2 h. DM-130 resin pretreatment method is the same as embodiment 1.

[0043] c. Separation and storage of immobilized lipase: the wet immobilized enzyme was separated by filtration and placed at 4°C for 24 hours.

[0044] d. Using propanol as the immobilized enzyme treatment agent, add 1g of wet immobilized enzyme to 40ml of propanol, vibrate at 35°C and 150rpm for 1.5h, filter the treated immobilized enzyme, and place it at 4°C for 24h. Determination of enzyme activity.

[0045] e. Soybean oil transesterification: Mix 22.5g soybean oil, 11ml n-hexane and 1g organic solvent-treated immobilized enzyme, preheat at 30°C for 20min, ...

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Abstract

The invention provides a method for improving the activity and stability of immobilized lipase. The adsorption immobilized lipase is soaked in a polar organic solvent which can change the molecular conformation of the lipase and the adsorption state of a carrier on the lipase and serves as a treating agent for appropriate time to obtain the lipase with high activity and stability; and an immobilized carrier is selected from 40-mesh active carbon, silica gel or DM-130resin, and an organic compound is one of alcohol, ketone and ester and serves as the immobilized enzyme treating agent. The adsorption immobilized lipase is soaked in the organic compound treating agent, and the soaked immobilized lipase undergoes separating collection and drying treatment to obtain the immobilized lipase with improved catalytic activity and stability. The treated immobilized lipase can be used for transesterification, synthesis or hydrolysis of grease, and compared with a reference immobilized lipase, the enzyme activity is improved by 1.5 to 2.5 times, the half life period is prolonged by 3 to 8 hours, and the immobilized lipase can be used for 3 to 6 times more for catalyzing the transesterification of soybean oil. And by the method, the use performance of the immobilized enzyme is improved.

Description

technical field [0001] The invention belongs to the field of enzymatic chemistry, and relates to the technology of improving the activity and stability of enzyme by fixing on inorganic activated carbon, silica gel and organic resin, and also relates to the technology of catalytic esterification and transesterification of organic matter. Background technique [0002] Lipases are commonly used industrially to catalyze esterification or transesterification reactions in organic media. Using lipase as a catalyst to prepare biodiesel has mild reaction conditions, less alcohol consumption, easy recovery of glycerol, and no waste, which can solve the problems of difficult separation and high energy consumption of catalysts used in the production of biodiesel by chemical methods. The enzyme immobilization technology solves the problems that lipase is easy to agglomerate, difficult to recycle, and difficult to reuse during the catalytic reaction process, and improves the activity and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08
Inventor 孙玉梅杜凯
Owner DALIAN POLYTECHNIC UNIVERSITY
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