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Preparation method of immobilized lipase

A technology for immobilizing lipase and porcine pancreatic fat, which is applied in the direction of immobilization on/in the organic carrier, can solve the problems of affecting mass transfer, blocking enzyme entry, hindering cross-linking reaction, etc., to improve the stability of use, The effect of long service life and low price

Inactive Publication Date: 2013-07-03
JIANGXI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One reason is that part of the aldehyde groups are inside the canvas, and intramolecular hydrogen bonds are formed between adjacent hydroxyl groups after encountering water, which blocks the contact between the aldehyde groups and the enzyme protein; the other reason is that after the dense fibers are filled with water and swell, the surface water The electrostatic tension of the membrane blocks the entry of macromolecules such as enzymes and affects mass transfer
Moreover, some adsorbent substances that are not easy to remove in the dry powder enzyme also adhere to the surface of the fiber group, hindering the progress of the crosslinking reaction.

Method used

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  • Preparation method of immobilized lipase
  • Preparation method of immobilized lipase
  • Preparation method of immobilized lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The present invention selects natural fiber---ramie as carrier, and three steps of activation of ramie, porcine pancreas lipase liquid preparation and cross-linking are carried out:

[0038] (1) Activation of ramie: cut some ramie, boil with 0.5mol / L KOH for 0.5 hour to remove impurities on the surface of ramie, then oxidize with 0.1 mol / L NaI04 solution, take it out after 6 hours and wash it with deionized water , add glycerol to the surface of ramie and let it stand for 40 min to remove unreacted NaIO4, and wash it with deionized water for later use.

[0039] (2) Preparation of enzyme solution: Dissolve a certain amount of porcine pancreatic lipase dry powder in 50mL of phosphate buffer (0.025mol / L, pH 7.5), stir evenly at 4°C, and centrifuge at 10,000r / min for 10 min , and then take the supernatant and store it in the refrigerator for later use.

[0040] (3) Cross-linking: put the activated ramie in step (1) in an Erlenmeyer flask and cross-link with the enz...

Embodiment 2

[0042] The present invention selects natural fiber---ramie as carrier, and three steps of activation of ramie, porcine pancreas lipase liquid preparation and cross-linking are carried out:

[0043] (1) Activation of ramie: cut some ramie, boil with 0.8mol / L KOH for 0.6 hours to remove impurities on the surface of ramie, then oxidize with 0.3 mol / L NaI04 solution, take it out after 8 hours and wash it with deionized water , add glycerol to the surface of ramie and let it stand for 50 min to remove unreacted NaIO4, and wash it with deionized water for later use.

[0044] (2) Preparation of enzyme solution: Dissolve a certain amount of porcine pancreatic lipase dry powder in 50mL of phosphate buffer (0.0.500mol / L, pH 7.5), stir evenly at 4°C, and centrifuge at 10,000r / min for 10 min, and then take the supernatant and store it in the refrigerator for later use.

[0045] (3) Cross-linking: Put the activated ramie in step (1) in a conical flask and cross-link with the enzym...

Embodiment 3

[0047] The present invention selects natural fiber---ramie as carrier, and three steps of activation of ramie, porcine pancreas lipase liquid preparation and cross-linking are carried out:

[0048] (1) Activation of ramie: Cut some ramie, boil with 1.0mol / L KOH for 1.0 hour to remove impurities on the surface of ramie, then oxidize with 0.6 mol / L NaI04 solution, take it out after 12 hours and wash it with deionized water, Add glycerol and let it stand for 60 min on the surface of ramie to remove unreacted NaIO4, and wash it with deionized water for later use.

[0049] (2) Preparation of enzyme solution: Dissolve a certain amount of porcine pancreatic lipase dry powder in 50mL of phosphate buffer solution (0.025-0.500mol / L, pH 7.5), stir evenly at a low temperature of 4°C, and dissolve at a speed of 10000r / min Centrifuge for 10 min, then take the supernatant and store it in the refrigerator for later use.

[0050] (3) Cross-linking: put the activated ramie in step (1) ...

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Abstract

The invention relates to a preparation method of immobilized lipase and is characterized in that natural herbaceous perennial plants, especially natural fiber-ramie and porcine pancreas lipase are used as raw materials, ramie is used as the carrier of the lipase. The preparation method comprises the following steps: activating ramie, preparing porcine pancreas lipase solution, and then adopting the covalent crosslinking method to perform the immobilization of the lipase. The ramie has wide sources and low cost. The immobilized lipase is used for catalysis and used to prepare biodiesel and can be used repeatedly; and the operational stability and catalytic effect of the immobilized lipase can be greatly increased and the production cost of biodiesel can be reduced.

Description

technical field [0001] The invention relates to a preparation method of immobilized lipase. technical background [0002] The covalent binding between the enzyme and the carrier is based on a chemical reaction between reactive amino acid residues on the surface of the enzyme (A) and reactive functional groups on the surface of the carrier (B). In order to achieve effective covalent linkage, relevant functional groups on the carrier or enzyme must be activated in advance. Generally, relevant functional groups on the carrier are activated in advance, and then covalently bonded with the enzyme. In a few cases, such as when it is necessary to control the binding method or the number of covalent bonds, the relevant functional groups on the enzyme molecule are activated in advance, and then covalently bonded to the carrier. [0003] After the enzyme is immobilized, it can not only maintain the catalytic activity of the enzyme but also overcome some shortcomings of the free enzym...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/02
Inventor 刘光斌黄长干刘苑秋胡冬南张露杜天真黄忠邱自兵
Owner JIANGXI AGRICULTURAL UNIVERSITY