Preparation method of immobilized lipase used for grease hydrolysis

An immobilized lipase, oil hydrolysis technology, applied in directions such as immobilization on/in organic carriers, fermentation, etc., can solve the problems of increasing environmental control costs, unbearable actual production, high price, etc. The effect of low cost and simple process

Inactive Publication Date: 2009-09-23
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The coupling agent bisacyl chloride-terminated polyethylene glycol used in the existing process is expensive, and the actual production is unbearable; the acid chloride is easily hydrolyzed to produce acid mist, which is harmful to the environment and further increases the cost of environmental prevention and control

Method used

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  • Preparation method of immobilized lipase used for grease hydrolysis
  • Preparation method of immobilized lipase used for grease hydrolysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1) Dissolve cellulose acetate in a dimethylacetamide / acetone mixture with a volume ratio of 3:2, stir and dissolve at room temperature to form a homogeneous solution with a concentration of 130 g / L, and let stand to remove air bubbles. obtain spinning solution;

[0024] 2) carry out electrospinning with the spinning solution that makes in step 1), spinning flow rate 1mL / h, spinning voltage is 17kV, and the distance from nozzle to receiving plate is 15cm, makes cellulose acetate superfine fiber film, SEM photo see figure 1 ;

[0025] 3) Place the cellulose acetate microfiber membrane in an ethanol solution of potassium hydroxide with a concentration of 0.5M, soak for 3 hours, take out the membrane after the reaction is completed, rinse it with deionized water several times, and dry it for use;

[0026] 4) Soak the cellulose acetate microfiber membrane obtained in step 3) with absolute ethanol, then immerse in epichlorohydrin, take it out after reacting at 37°C for 3 ho...

Embodiment 2

[0030] The steps are the same as in Example 1, but in step 1), cellulose acetate butyrate is used instead of cellulose acetate, and the enzyme loading is 17 mg enzyme / g membrane. Electron micrographs of cellulose acetate butyrate microfiber membranes are shown in figure 2 .

[0031] The prepared cellulose acetate butyrate microfiber membrane immobilized lipase was used for soybean oil hydrolysis, the activity was 267U / g immobilized enzyme; for castor oil hydrolysis, the activity was 256U / g immobilized enzyme; For the hydrolysis of seed oil, the activity is 268U / g immobilized enzyme; for the hydrolysis of peanut oil, the activity is 264U / g immobilized enzyme; for the hydrolysis of cottonseed oil, the activity is 237U / g immobilized enzyme; for the hydrolysis of rice bran oil, the activity is 248U / g immobilized enzyme; for algae oil hydrolysis, the activity is 249U / g immobilized enzyme; for the mixed oil hydrolysis of soybean oil and castor oil with a volume ratio of 1:1, the a...

Embodiment 3

[0033]1) Dissolve cellulose acetate butyrate in a dimethylacetamide / acetone mixture with a volume ratio of 1:1, stir and dissolve at room temperature to form a uniform solution with a concentration of 20g / L, and let it stand to remove air bubbles to obtain spinning solution;

[0034] 2) Electrospinning is carried out with the spinning solution prepared in step 1), the spinning flow rate is 5mL / h, the spinning voltage is 10kV, and the distance from the nozzle to the receiving plate is 10cm, and cellulose acetate butyrate superfine fibers are obtained membrane;

[0035] 3) Place the cellulose acetate butyrate ultrafine fiber membrane in an ethanol solution of potassium hydroxide with a concentration of 1M, soak for 1 hour, take out the membrane after the reaction is completed, rinse it several times with deionized water, and dry it for use;

[0036] 4) Soak the cellulose acetate butyrate superfine fiber membrane obtained in step 3) with absolute ethanol, then immerse in epichlo...

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Abstract

The invention discloses a preparation method of immobilized lipase used for grease hydrolysis, comprising the following steps of: dissolving cellulose in a dimethyl acetamide/acetone mixed solution to prepare a uniform solution which is rested for removing bubbles to obtain a spinning solution; then preparing cellulose superfine fibrous membrane by electrostatic spinning; placing the cellulose superfine fibrous membrane in the ethanol solution of potassium hydroxide for hydrolysis to generate hydroxyl group which is activated after being washed for a plurality of times by de-ionized water; and then placing the activated cellulose superfine fibrous membrane in an enzyme solution, and obtaining the immobilized lipase of the cellulose superfine fibrous membrane after reaction for a plurality of hours. The prepared immobilized lipase can be used for grease hydrolysis. The method adopts the cellulose superfine fibrous membrane and has the advantages of simple and fast technology, good repeatability, low cost and safe operation and being environment-friendly and suitable for large-scale industrialized production, furthermore, the prepared immobilized lipase has large enzyme-loading capacity, and catalytic efficiency for grease hydrolysis is high.

Description

technical field [0001] The invention relates to the technical field of immobilized enzymes, in particular to a preparation method of an immobilized lipase used for hydrolysis of fats and oils, which uses a cellulose superfine fiber membrane as an immobilized carrier. Background technique [0002] Lipase, also known as triglyceride hydrolase, is derived from animals, plants and microorganisms, and its natural substrate is triglyceride. It can catalyze a series of reactions such as alcoholysis of esters, ester synthesis, transesterification, lactone synthesis and amide synthesis. Lipase also has the characteristics of good substrate specificity, regioselectivity and stereoselectivity, making it of great application value in the chiral resolution of racemic mixtures and the asymmetric synthesis of drug intermediates. Because immobilized enzymes have a series of advantages such as easy continuous production, high enzyme stability, easy control of reaction conditions, easy produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12C12N11/06C12P7/64
Inventor 万灵书黄赋黄小军徐志康
Owner ZHEJIANG UNIV
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