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Carrier-free immobilized lipase and preparation method thereof

A carrier-free immobilization and lipase technology, applied in the direction of immobilized enzymes, biochemical equipment and methods, enzymes, etc., can solve the problems of high price and unfavorable industrial production of biodiesel, and achieve the effect of easy operation and low cost

Inactive Publication Date: 2011-08-17
SHENZHEN UNIV
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the lipase used for biodiesel research at home and abroad is mainly Novozym 435 produced by Novozymes. This enzyme is a resin-immobilized enzyme, and the carrier is a macroporous acrylic resin. The price is high, which is not conducive to the industrial production of biodiesel prepared by enzymatic method.

Method used

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  • Carrier-free immobilized lipase and preparation method thereof

Examples

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Effect test

Embodiment 1

[0035] Example 1 Preparation and catalytic hydrolysis of carrier-free immobilized Penicillium extensa lipase

[0036] Weigh 1.5g of PEL freeze-dried enzyme powder into a 500mL Erlenmeyer flask, add 300mL of 50% ammonium sulfate phosphate buffer (pH6.5, 0.05M), and stir magnetically at 4°C until the enzyme powder particles are completely dissolved; take 1mL of 50% glutaraldehyde Mix with 3mL phosphate buffer to get 12.5% ​​glutaraldehyde, add 3mL dropwise to the above-mentioned Erlenmeyer flask and react for 24 hours; after centrifugation, the supernatant has no enzyme activity, and the precipitate is washed with phosphate buffer until the supernatant has no color (Washed 8 times in total, and the eluate had no enzyme activity); vacuum dried overnight, and weighed 0.3977g.

[0037] Support-free immobilized Penicillium extensa lipase catalyzed hydrolysis reaction:

[0038] Take 30mg of immobilized enzyme for reaction, the reaction conditions are as follows:

[0039] Phosphate ...

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Abstract

The invention relates to carrier-free immobilized lipase and a preparation method thereof. The preparation method comprises the following steps of: 1, preparing enzyme solution; 2, precipitating lipase molecules by a buffer system with the final concentration of ammonium sulfate of 50 percent; 3, adding a cross-linking agent glutaraldehyde; 4, centrifuging to obtain water insoluble precipitate particles; and 5, freezing and drying the water insoluble precipitate particles to ensure the water insoluble precipitate particles to be dried completely so as to obtain a flavescent powder material, i.e. the carrier-free penicilllum expansum immobilized lipase (PEL-CLEA). In the method, the lipase molecules are cross-linked and aggregated by the cross-linking agent glutaraldehyde through the lipase under the action of ammonium sulfate precipitates, so that the purpose of immobilizing the lipase is fulfilled. The method has low cost and is easy to operate.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to a carrier-free immobilized lipase and a preparation method thereof. Background technique [0002] As a kind of biocatalyst, enzyme has been widely used in food production and testing, environmental protection technology, biotechnology and other fields in recent years. However, the disadvantages of enzymes, such as high price, poor stability, and difficulty in recovery after reaction, limit the development and application of enzyme preparation products. In this case, immobilized enzymes came into being. Enzyme immobilization is a kind of technology that binds or confines enzymes in a certain area with solid materials, and can still carry out its unique catalytic reactions, and can be recycled and reused. Compared with free enzymes, immobilized enzymes overcome the shortcomings of free enzymes while maintaining their high-efficiency, specificity and mild enzyme-catalyzed reacti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/00
Inventor 杨缜赖璟琦
Owner SHENZHEN UNIV
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