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TL immobilization enzyme and application thereof

A technology for immobilizing lipase and lipase, which is applied to the immobilized enzyme method and its application field, and can solve the problems of not producing soap, unable to meet production requirements, producing soap and the like

Active Publication Date: 2012-12-26
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, so far, the existing TL-immobilized enzymes on the market cannot meet the production requirements
In the transesterification reaction it catalyzes, it is found that there is a large amount of soap in the product, which wastes a lot of raw materials and increases the cost. The reason may be that there are some insurmountable technical problems in the process of combining different types of carriers and enzymes, or that the carriers and enzymes are immobilized. Conflicts with the reaction medium during the process
Therefore, the current immobilized TL requires a large amount of oil to be eluted, resulting in a large amount of waste of raw materials
At the same time, it brings great difficulties to the downstream separation and purification, which limits its application in the oil industry.
[0009] Therefore, there is an urgent need to develop a new immobilized TL enzyme method in this area, so that the TL enzyme maintains a high activity and does not produce soap in the reaction, thereby expanding the application range of TL lipase in the food industry and overcoming the existing immobilization method. Problems of Soap Production by NTL Enzyme

Method used

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  • TL immobilization enzyme and application thereof
  • TL immobilization enzyme and application thereof
  • TL immobilization enzyme and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0092] An exemplary preferred embodiment of the present invention is as follows:

[0093] Step 1. Carrier processing

[0094] Soak the macroporous adsorption resin carrier in deionized water to remove impurities, then soak in ethanol overnight to remove organic impurities and other insoluble matter, rinse with deionized water until there is no alcohol smell, and then soak in deionized water for storage.

[0095] Alternatively, wash the ion exchange resin with deionized water, alternately treat it with 5% NaOH and 5% HCl for 2 to 5 times, and then treat it with 5% NaOH for about two hours each time, and then treat it with deionized water until it reaches water level. Neutral, soak in deionized water for later use.

[0096] Step 2. Enzyme immobilization

[0097] Weigh 40g of the treated resin carrier into a 250ml Erlenmeyer flask, put 40ml of TL enzyme solution into it, and fix it in a water-bath shaker at 25°C at 150rpm. During this period, take the supernatant at regular int...

Embodiment 1

[0112] Embodiment 1. Adopt macroporous adsorption resin to prepare immobilized enzyme

[0113] The macroporous adsorption resins D101, HPD100A or X-5 (D101 and HPD100A were purchased from Hebei Cangzhou Baoen Chemical Co., Ltd., and X-5 was purchased from Shanghai Huazhen Resin Co., Ltd.) were soaked in deionized water for about 12 hours to remove impurities, Then soak in 95% ethanol overnight to remove organic impurities and other insoluble matter, rinse with deionized water until there is no alcohol smell, and place in deionized water for later use.

[0114] Prepare immobilized enzyme 1-19 according to the following formula and steps:

[0115] 1. Weigh 40g of the processed D101 resin into a 250ml Erlenmeyer flask, add 40ml pH5 TL enzyme solution, shake at 150rpm in a shaker at 25°C for 2h, then take out the resin and put it into a dry petri dish, and put it into a freeze-dried Drying is carried out in the machine, and the specific freeze-drying parameters are set as:

[01...

Embodiment 2

[0149] Embodiment 2. adopt ion exchange resin to prepare immobilized enzyme

[0150] Styrene-DVB201 × 2 resin (purchased from Tianjin Nankai Hecheng Technology Co., Ltd.) was first washed with deionized water, then alternately soaked twice with 5% NaOH and 5% HCl, and then soaked once with 5% NaOH. The second treatment is about two hours, and then washed with deionized water until the water is neutral, soaked in deionized water for later use.

[0151] Prepare immobilized enzyme 20-24 according to the following formula and steps:

[0152] 1. Take out 40g of the treated Styrene-DVB201×2 resin into a 250ml Erlenmeyer flask, add 40ml of pH5 TL enzyme solution, and 0.8ml of 1% glutaraldehyde solution, shake at 150rpm for 2h in a shaker at 25°C, and then take out the resin Put it into a dry petri dish, put it in a well-ventilated place for natural air-drying, and obtain immobilized enzyme 20 after drying.

[0153] 2. Take out 40g of the processed Styrene-DVB201×2 resin into a 250m...

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Abstract

The present invention relates to a TL immobilization enzyme and an application thereof. Specifically the present invention provides a preparation method for an immobilization lipase. The method comprises the following steps: a) carrying out a pre-treatment on a carrier, wherein the carrier is selected from an ion exchange resin or a macroporous adsorption resin having a polystyrene skeleton; b) providing a lipase; c) carrying out contact of the pre-treated carrier from the step a) and the lipase provided by the step b), such that the lipase is immobilized on the pre-treated carrier; and d) drying. The present invention further provides the immobilization lipase prepared by using the method of the present invention and an application thereof. According to the present invention, the immobilization enzyme of the present invention has high activity and stability, and does not generate soap during a process of catalysis of an ester exchange reaction in food industry, operation is simple, and oil processing cost is substantially reduced.

Description

technical field [0001] The invention belongs to the fields of food and chemical industry. More specifically, the present invention relates to an enzyme immobilization method for TL lipase and its application. Background technique [0002] As a biocatalyst, enzyme has the advantages of substrate selectivity, mild reaction conditions, and less pollution. It overcomes the disadvantages of harsh reaction conditions and environmental pollution caused by reaction by-products brought about by traditional industrial catalysts, and has good application potential. Finding enzymes with high catalytic activity and mild reaction conditions, or modifying existing enzymes to make them more suitable for production applications, is the main development direction of enzyme engineering in industrial applications in the future. [0003] Immobilized enzyme technology is a technology that binds or restricts free enzymes to a certain area by physical or chemical means, so that they can still perf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12P7/64
Inventor 郑妍辛本荣杨天奎徐学兵
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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