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Immobilization lipase using loofah sponge as carrier and preparation method thereof

A technology of immobilizing lipase and loofah, which is applied in the direction of immobilizing on/in organic carriers, can solve the problems of high cost, large-scale application of restriction enzyme method, low enzymatic activity of immobilized enzyme, etc., and achieve repeated use High efficiency, good mechanical strength and stability, easy to obtain effect

Inactive Publication Date: 2008-09-10
SHANGHAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the immobilized enzyme obtained by the above carrier is either too low in enzyme activity or too high in cost, which limits the large-scale application of enzymatic method in production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment one: concrete steps are as follows:

[0033] 1. Treatment of loofah: Wash the dried loofah with 0.5mol / L NaOH solution at 70-80°C for 12 hours to remove excess oil. Then the loofah was washed 3 times with deionized water, and then the loofah was soaked in deionized water and washed with ultrasonic waves for 3 hours. After drying at room temperature, cut the loofah into 1cm×3cm size for later use.

[0034] 2. Enzyme purification: Dissolve the thick Candida cylindrical powder in 0.1mol / L phosphate buffer solution with pH=8, centrifuge at 3000rpm / min for 15min, take the supernatant, then freeze-dry the supernatant into powder spare.

[0035] 3. Immobilization of enzyme: 4% (wt) loofah carrier and 95.5% (wt) cyclohexane mix uniformly, keep the constant temperature in a shaker at 25°C for 1h, then add 0.5% (wt) lipase powder, mix uniformly, Fix at 25°C for 16h. The immobilized carrier was taken, washed three times with cyclohexane, and dried at room temperatur...

Embodiment 2

[0037] Embodiment two: this embodiment is basically the same as embodiment one, the difference is that the organic solvent adopted is n-hexane, and the lipase adopted is: Aspergillus niger enzyme. As a result, it was found that the activity recovery rate of the obtained immobilized enzyme reached 76%.

Embodiment 3

[0038] Embodiment 3: This embodiment is basically the same as Embodiment 1, except that the organic solvent used is n-heptane. The lipase used is: Pseudomonas fluorescens. It was found that the activity recovery rate of the immobilized enzyme reached 81%.

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Abstract

The invention relates to an immobilized lipase which takes fasciculus vascularis luffae as a carrier and preparation of the immobilized lipase. In the immobilized lipase, the mass ratio of the carrier lipase and the fasciculus vascularis luffae is between1:4 and 1:8. The concrete preparation method comprises deoiling treatment of the fasciculus vascularis luffae, refinement of the lipase and immobilization of the lipase. The invention takes the fasciculus vascularis luffae as the carrier for immobilizing the lipase, thereby the immobilized lipase obtained has high activity recovery rate and high repeated utilization ratio and can be used for a plurality of times without deactivation. Moreover, the technological method is simple in operation and has a chance of promoting the industrial progress for enzymatical production of biological diesel fuels.

Description

technical field [0001] The invention relates to an immobilized lipase and a preparation method thereof, in particular to an immobilized lipase with loofah as a carrier and a preparation thereof. Background technique [0002] Enzymes have always catalyzed reactions in the free state dissolved in water or in intact cells. However, even if the free enzymes are catalyzed under the most suitable conditions for the enzyme reaction, they are easily inactivated and cannot be recovered after the reaction. Defects, sometimes difficult to separate from the product, affect the purity and quality of the product. [0003] Compared with general water-soluble enzyme preparations, immobilized enzymes not only still have the characteristics of high specificity of enzymes, catalytic efficiency and mild action conditions, but also are more stable than water-soluble enzymes and have a longer service life. Generally, they can be used continuously for several days. Hundreds of hours or even month...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04
Inventor 张宝华潜飞杨青叶俊丹叶正尧
Owner SHANGHAI UNIV
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