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Method for immobilizing Thermomyces lanuginosus lipase

A technology of thermomyces and lipase, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of high price and insufficient chemical methods of commercial lipase, and achieve convenient operation and simple preparation conditions Effect

Inactive Publication Date: 2017-08-04
ZHEJIANG UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the problem of high price of commercialized lipase and the deficiencies of chemical methods, and to prepare immobilized Thermomyces lanuginosa lipase by means of cross-linking immobilization method, which has not been reported in the literature

Method used

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  • Method for immobilizing Thermomyces lanuginosus lipase
  • Method for immobilizing Thermomyces lanuginosus lipase
  • Method for immobilizing Thermomyces lanuginosus lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Add 100ml of liquid lipase Lipozyme TL 100L (Thermomyces lanuginosa lipase, Novozymes, Denmark) to 400ml of tert-butanol at 4°C, let it settle for 1 hour, and centrifuge at 10,000rpm for 10 minutes. Remove the supernatant, add 20g of non-polar macroporous adsorption resin D101 and 20ml of crosslinking agent (5% polyethyleneimine aqueous solution) for precipitation, shake the shaker for 2 hours, and then dry the reaction solution in vacuum at 45°C for 4 hours , to prepare 9.96 g of immobilized lipase. The esterification activity of the immobilized lipase was measured, and the specific enzyme activity was 3580U / g.

[0027] The definition and determination method of the esterification activity unit of immobilized lipase are as follows:

[0028] Definition of lipase esterification activity (U): the amount of enzyme required to generate 1 μmoL of the product methyl oleate per minute at 50°C. The specific enzyme activity is calculated as follows: X=[V×(V1 / V2)]ρ / [(m T) M]

...

Embodiment 2

[0033] Prepare different batches of immobilized lipase according to the method of Example 1, and use the esterification activity of different batches of commercial liquid lipase Lipozyme TL 100L as a control to obtain the batch stability comparison results of immobilized lipase, as shown in Table 1 The results showed that the enzyme activity and stability of the self-made immobilized lipase were much higher than the commercial enzyme Lipozyme TL IM.

[0034] Table 1: The stability comparison results of the immobilized lipase prepared in Example 1 and commercialized enzyme

[0035]

Embodiment 3

[0037] The organic solvent of Example 1 is changed to that shown in Table 1, and other conditions are unchanged. The esterification activity of the obtained immobilized lipase was determined, and the results are shown in Table 2.

[0038] Table 2: Specific enzyme activity results of precipitation with different organic solvents

[0039]

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Abstract

The invention discloses a method for immobilizing Thermomyces lanuginosus lipase. The method comprises the following steps: adding 100 L of an enzyme liquid of Thermomyces lanuginosus lipase Lipozyme TL into an organic solvent at 4 DEG C, carrying out standing precipitation for 1 h, centrifuging the obtained solution, taking the obtained supernatant, adding a cross-linking agent and macro-porous adsorption resin, carrying out oscillation cross-linking immobilization, and carrying out vacuum drying after the above reaction is completed in order to obtain immobilized Thermomyces lanuginosus lipase. The immobilized lipase has the advantages of simple preparation conditions, and convenience in operation. The enzyme activity and the batch stability of the immobilized Thermomyces lanuginosus lipase are much higher than those of the commercialized enzyme Lipozyme TL IM, so the method is suitable for industrial application.

Description

[0001] (1) Technical field [0002] The invention relates to a preparation method of immobilized Thermomyces lanuginosa lipase and its application to the methyl esterification reaction of soybean oil deodorization distillate. [0003] (2) Background technology [0004] Lipase (Lipase, EC3.1.1.3, glyceride hydrolase) is an enzyme that decomposes fat. It is ubiquitous in animals, plants and microorganisms. It can catalyze the decomposition, esterification, transesterification, lactone synthesis, and peptide synthesis of ester compounds. Synthesis, ester polymerization and acylation reactions are among the most studied and applied enzymes. The reaction catalyzed by lipase in the organic phase can synthesize a variety of high-value products, such as optically active drugs, structured lipids with special physiological functions, fine chemicals, chiral compounds, and biodiesel, which have great application value. [0005] The most prominent feature of lipase is that it can catalyze ...

Claims

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Application Information

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IPC IPC(8): C12N11/08
Inventor 郑建永汪钊韦威章银军王升帆许新德
Owner ZHEJIANG UNIV OF TECH
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