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
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[0013] Example 1:
[0014] (1) Weigh 1g block SiO 2 The macroporous material was dried at 80℃ for 2h, and placed in 50ml, 0.3mol·L -1 Sodium aluminate solution in a hydrothermal reactor with polytetrafluoroethylene lining, react at 120°C for 12 hours, after natural cooling, filter out with 8-10 mesh screen, rinse with deionized water repeatedly, and change after drying SiO 2 Macroporous material Al-SiO 2 .
[0015] (2) Al-SiO 2 Immerse in saturated copper acetate solution and shake for 4 hours, filter out, repeatedly rinse with deionized water, and dry to obtain the carrier Cu for immobilized enzyme 2+ -Al-SiO 2 .
[0016] (3) Weigh 1g Cu 2+ -Al-SiO 2 , Immersed in 50ml with pH=4.5 buffer solution, the protein concentration is 0.1mg·ml -1 In the crude laccase solution at 25℃, rotating speed 50r·min -1 Shake in a shaker for 5 hours, filter with 8-10 mesh, and wash with buffer until no protein can be detected in the supernatant to obtain immobilized laccase. The recovery rate of the e...
Example Embodiment
[0017] Example 2:
[0018] (1) Same as Example 1(1).
[0019] (2) Al-SiO 2 Immerse 0.5mol·L -1 Shake in zinc acetate solution for 5 hours, filter out, repeatedly rinse with deionized water, and dry to obtain carrier Zn for immobilized enzyme 2+ -Al-SiO 2 .
[0020] (3) Weigh 0.1g Zn 2+ -Al-SiO 2 , Immersed in 5ml and configured with pH=4.0 buffer solution, the protein concentration is 0.05mg·ml -1 In the crude laccase solution at 25℃, rotating speed 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 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 is 71.2% of the initial enzyme activity.
Example Embodiment
[0021] Example 3:
[0022] (1) Same as Example 1(1).
[0023] (2) Al-SiO 2 Immerse 0.5mol·L -1 Shake in ferric chloride solution for 4 hours, filter out, repeatedly rinse with deionized water, and dry to obtain carrier Fe for immobilized enzyme 3+ -Al-SiO 2 .
[0024] (3) Weigh 1g Fe 3+ -Al-SiO 2 , Immersed in 50ml with pH=4.0 buffer solution, the protein concentration is 0.05mg·ml -1 In the crude laccase solution at 25℃, rotating speed 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 27.5 U·g -1 , After 10 batches of repeated operations with the substrate ABTS, the remaining enzyme activity is 77.4% of the initial enzyme activity.
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