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SiO2 based macroporous material immobilized laccase and preparation method thereof

A technology for immobilizing laccase and macroporous materials, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of difficult industrial application, complex processing technology, poor separation effect, etc., and achieve stable properties and simple preparation process , High recovery effect

Inactive Publication Date: 2014-12-17
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing immobilized laccases generally have disadvantages such as high production cost, complicated processing technology, and poor separation effect, so it is difficult to apply industrially

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] (1) Weigh 1g bulk SiO 2 The macroporous material was dried at 80°C for 2 hours, placed in a solution containing 50ml, 0.3mol·L -1 The sodium aluminate solution is placed in the polytetrafluoroethylene lining of the hydrothermal reaction kettle, reacted at 120°C for 12 hours, and after natural cooling, filter it out with a 8-10 mesh screen, rinse it repeatedly with deionized water, and obtain the improved product after drying. Sex SiO 2 Macroporous material Al-SiO 2 .

[0015] (2) Al-SiO 2 Immerse in a saturated copper acetate solution and shake for 4 hours, filter out, rinse repeatedly with deionized water, and dry to obtain the carrier Cu for immobilized enzymes. 2+ -Al-SiO 2 .

[0016] (3) Weigh 1g Cu 2+ -Al-SiO 2 , immerse in 50ml and use pH=4.5 buffer solution to prepare a protein concentration of 0.1mg·ml -1 In the crude laccase solution, at 25°C, the rotation speed is 50r·min -1 Shake in a shaker for 5 hours, filter out with an 8-10 mesh screen, wash wit...

Embodiment 2

[0018] (1) With embodiment 1 (1).

[0019] (2) Al-SiO 2 Immerse in 0.5mol·L -1 Shake in a zinc acetate solution for 5 hours, filter out, rinse repeatedly with deionized water, and dry to obtain the carrier Zn for immobilized enzymes. 2+ -Al-SiO 2 .

[0020] (3) Weigh 0.1g Zn 2+ -Al-SiO 2 , immerse in 5ml of buffer solution with pH=4.0, the protein concentration is 0.05mg·ml -1 In the crude laccase solution, at 25°C, the rotation speed is 50r·min -1 Shake in a shaker for 5 hours, filter out, and wash with buffer until no protein can be detected in the supernatant to obtain immobilized laccase. The recovery rate of its enzyme activity is 95.4%, and the enzyme activity is 20.0U·g -1 , after 10 batches of repeated operations with the substrate ABTS, the remaining enzyme activity was 71.2% of the initial enzyme activity.

Embodiment 3

[0022] (1) With embodiment 1 (1).

[0023] (2) Al-SiO 2 Immerse in 0.5mol·L -1 Shake in ferric chloride solution for 4 hours, filter out, rinse repeatedly with deionized water, and dry to obtain the carrier Fe for immobilized enzyme 3+ -Al-SiO 2 .

[0024] (3) Weigh 1g Fe 3+ -Al-SiO 2 , immerse in 50ml and use pH=4.0 buffer solution to prepare protein concentration of 0.05mg·ml -1 In the crude laccase solution, at 25°C, the rotation speed is 50r·min -1 Shake in a shaker for 5 hours, filter out, and wash with buffer until no protein can be detected in the supernatant to obtain immobilized laccase. The recovery rate of the enzyme activity is 96.8%, and the enzyme activity is 27.5U·g -1 , after 10 batches of repeated operations with the substrate ABTS, the remaining enzyme activity was 77.4% of the initial activity.

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Abstract

The invention relates to a SiO2 based macroporous material immobilized laccase and a preparation method thereof, and belongs to the technical field of immobilized laccase preparation. The immobilized laccase is prepared by fixing laccase on a carrier through a static and coordination effect, wherein the carrier is a SiO2 macroporous block material whose surface is modified. The preparation method comprises two steps namely carrier preparation and laccase immobilization, wherein the carrier preparation comprises the following steps: modifying a SiO2 macroporous block material by sodium aluminate under a hydrothermal condition, and then carrying out ion exchange to obtain a finished carrier product containing multivalent metal ions on the surface; and the laccase immobilization comprises the following steps: soaking the prepared carrier in a laccase solution, carrying vibration absorption at a room temperature, washing, and drying so as to obtain the finished immobilized laccase product. The provided immobilized laccase has the characteristics of simple preparation technology, low cost, high activity recycling rate, and stable property.

Description

technical field [0001] The invention belongs to the technical field of immobilized enzyme preparation, in particular to a SiO 2 Immobilized laccase based on macroporous material and preparation method thereof. Background technique [0002] Laccase (Laccase, E.C.1.10.3.2) is a copper-containing polyphenol oxidase, widely present in a variety of organisms. Laccase has broad substrate specificity and good stability, and can catalyze the oxidation of a variety of refractory organic pollutants, including phenolic compounds, polycyclic aromatic hydrocarbons, polychlorinated biphenyls and their derivatives, herbicides, It has important application value in wastewater treatment, biosensor construction, biopulping, food processing and organic synthesis. Therefore, scientists at home and abroad have carried out extensive research work on laccase in many fields such as biology, chemistry, physics, and medicine. [0003] The application of free laccase is limited because it is easy t...

Claims

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Application Information

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IPC IPC(8): C12N11/14
Inventor 梁云霄张群张瑞丰张育淇刘晓贞刘燕金林铃
Owner NINGBO UNIV
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