Magnetic composite microsphere immobilized laccase and preparation method thereof
A technology of magnetic composite microspheres and immobilized laccase, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of large loss of enzyme activity, large loss of enzyme activity, and shedding, so as to avoid chemical damage and benefit Effects of recovery and reuse, improvement of operational stability
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[0022] Example 1
[0023] A preparation method of immobilized laccase on magnetic composite microspheres mainly includes two steps:
[0024] Vector preparation
[0025] (1) Take FeCl with a molar ratio of 1:2 2 ·4H 2 O and FeCl 3 ·6H 2 The mixed powder of O was dissolved in 50 mL of a 0.5% sodium dodecyl sulfonate solution, and a 2 mol / l NaOH solution was added dropwise with stirring to adjust the pH to about 12. After the obtained product is matured in a water bath at 80°C for 30 minutes, it is repeatedly washed with distilled water to neutrality, dried and ground at 50°C to obtain ferroferric oxide powder;
[0026] (2) Take the ground Fe 3 O 4 1.5g was ultrasonically dispersed in 60ml of secondary water, while 6.4ml of tetraethoxysilane was ultrasonically dispersed in 120ml of methanol, and the methanol solution of tetraethoxysilane was added to Fe under mechanical stirring. 3 O 4 After stirring vigorously for 15 minutes, add 6ml of concentrated ammonia water, continue to stir the ...
Example Embodiment
[0031] Example 2
[0032] The preparation method of the carrier is as in Example 1, except that the concentration of polyacrylamide is 0.5 mg / L, the concentration of copper sulfate solution is 0.1 mol / L, and the shaking time is 12 hours. The laccase immobilization method is the same as in Example 1, except that 0.1g of the carrier product and 5mL of free laccase dissolved in disodium hydrogen phosphate-citric acid buffer solution are thoroughly mixed, and then stirred at pH 3.0 and temperature 5℃. hour. The mixing ratio of free laccase and carrier product is 20mg / 1g.
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[0033] Example 3
[0034] The carrier preparation method is as in Example 1, except that the polyacrylamide concentration is 5 mg / L, the copper sulfate solution concentration is 0.05 mol / L, and the shaking time is 8 hours; the laccase fixation method is the same as in Example 1, the difference is 0.1 After the finished product of g carrier is fully mixed with 5 mL of laccase dissolved in disodium hydrogen phosphate-citric acid buffer solution, the mixture is stirred for 5 hours under the conditions of pH 5.0 and temperature 15°C. The mixing ratio of free laccase and carrier product is 50mg / 1g.
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