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Preparation method of immobilized lipase

A technology for immobilizing lipase and lipase, applied in the direction of being immobilized on/in an organic carrier, can solve the problems of affecting mass transfer, blocking enzyme entry, hindering cross-linking reaction, etc. Long service life, low price effect

Inactive Publication Date: 2011-09-21
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
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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 enzyme solut...

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 enzyme soluti...

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) in a con...

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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 present invention relates to a preparation method of immobilized lipase. technical background [0002] The covalent bonding between the enzyme and the carrier is based on a chemical reaction, that is, the reaction between the reactive amino acid residues on the enzyme surface (A) and the reactive functional groups on the carrier surface (B). In order to achieve effective covalent attachment, the relevant functional groups on the carrier or enzyme must be activated in advance. In general, the relevant functional groups on the carrier are activated in advance, and then covalently bound to the enzyme. In a few cases, when it is necessary to control the binding mode or the number of covalent bonds, the relevant functional groups on the enzyme molecule are activated in advance, and then covalently binds to the carrier. [0003] After the enzyme is immobilized, it can not only maintain the catalytic activity of the enzyme, but also overcome some defici...

Claims

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Application Information

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Patent Type & Authority Applications(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
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