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Immobilized enzyme and preparing method thereof

A technology for immobilizing enzymes and enzyme proteins, which is applied in the direction of immobilizing on or in inorganic carriers, or in or on organic carriers, and can solve problems such as poor specificity, poor random transesterification activity, and lack of

Inactive Publication Date: 2017-06-09
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of immobilized enzymes prepared by two different carriers is quite different. RM IM has specificity and poor random transesterification activity, while TL IM has high random transesterification activity but poor specificity.
At present, there is a lack of an immobilized enzyme method, which can make the immobilized lipase product have high transesterification activity and specificity at the same time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0119] Preparation method of immobilized enzyme

[0120] Another object of the present invention is to provide the preparation method I of immobilized enzyme, it comprises

[0121] (1) The step of mixing the hydrophobic carrier with the enzyme,

[0122] (2) A step of adding a hydrophilic carrier and an enzyme to the mixture obtained in step (1).

[0123] In the above step (1), the mixing ratio of hydrophobic carrier and enzyme is calculated by weight ratio, hydrophobic carrier: enzyme is 1-100:1, preferably 10-90:1, more preferably 20-80:1, even more preferably It is 30-70:1, most preferably 40-60:1, wherein the amount of enzyme is calculated by enzyme protein.

[0124] In a specific embodiment of the present invention, the mixing ratio of the hydrophobic carrier and the enzyme is calculated by weight ratio, and the ratio of hydrophobic carrier: enzyme is 120:2.2, wherein the amount of the enzyme is calculated by enzyme protein.

[0125] In a preferred embodiment of the pre...

Embodiment 1

[0196] Weigh the treated hydrophobic resin sepabeads EC-HA (Mitsubishi Chemical) 120g, add lipase TL ( Thermomyces lanuginose ) enzyme solution (activity unit is 1000KLU / g ， The protein content in the enzyme liquid is 2% by weight) 110 ml (Novozymes), placed in a shaker at 20°C for 24 h, then took out the filtered resin, dried at room temperature until the moisture content was below 10%, and placed the sample in a high-speed In the mixing granulator (model: GHL 5, Changzhou Jiafa Granulating Equipment Co., Ltd.), add 50g of hydrophilic K160 white carbon black (Yihai Kerry Jiamusi Factory), and add 20ml of lipase enzyme solution to another 100ml of water ( Novozymes, trade name lipozyme TL 100L), 3g sodium carboxymethyl cellulose, mixed and dissolved evenly, sprayed into the high-speed mixing granulator. When the resin-mixed silica particles become larger to 450-800 microns, the granulation is stopped, and the samples are taken out for shot blasting in a shot blasting machin...

Embodiment 2

[0199] Take 40g of Aerosil R974 hydrophobic silica (Degussa Company), 140g of K160 hydrophilic silica (Yihai Jiali Jiamusi factory), mix them evenly, and place them in a high-speed mixing granulator (GHL 5 manufactured by Changzhou Jiafa Co., Ltd. Granular Equipment Company), another 200ml enzyme solution (Novozymes, trade name lipozyme TL 100L, activity unit is 1000KLU / g ， The protein content in the enzyme solution is 2% by weight), 100ml of water, 6g of gelatin are dissolved and mixed evenly, and then pumped into the granulator at a uniform speed. After the granules are formed, the samples are taken out for shot blasting, and naturally dried to a moisture content below 8%. Immobilized enzyme 2 was obtained. Among them, hydrophilic carrier: hydrophobic carrier: enzyme protein = 140:40:4 (weight ratio).

[0200] Aerosil R974 has an average pore size of 5nm and a specific surface area of ​​170m 2 / g, the average particle size is 20 μm. K160 silica has an average pore size o...

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Abstract

The invention provides an immobilized enzyme and a preparing method thereof. The immobilized enzyme is characterized by comprising a lyophobic carrier, a hydrophilic carrier and zymoprotein, wherein relative to the total weight of the immobilized enzyme, the content of the lyophobic carrier is 0.1-99 wt%, the content of the hydrophilic carrier is 5-99 wt%, the content of enzyme is 0.05-10 wt%, and the content of enzyme is based on zymoprotein. The preparing method is characterized by comprising the steps of 1, mixing the lyophobic carrier with the enzyme; and 2, adding the hydrophilic carrier and the enzyme to the mixture obtained in step 1. The immobilized enzyme has high specificity and has esterification and ester exchange functions.

Description

technical field [0001] The invention relates to an immobilized enzyme having esterification, transesterification and Sn-1,3 selectivity simultaneously, a preparation method thereof and an esterification method using the immobilized enzyme. Background technique [0002] The full name of lipase (lipase, EC3.1.1.3) is triglyceride hydrolase. Lipases from different strains can also have functions such as esterification, transesterification, alcoholysis, and racemate resolution. Liquid lipase cannot be reused and is difficult to separate, which limits its industrial application. The above problems can be solved by immobilizing the enzyme on a carrier with certain physical properties. [0003] Lipase has a hydrophobic cavity in its structure and a lid on it, and the opening and closing of the lid is closely related to the hydrophobic / hydrophilic interface. The immobilized lipase will show different activities due to the influence of carrier properties. In the non-patent literat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/08C12N11/04
Inventor 辛本荣郑妍郑磊
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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