Magnetic immobilized spore laccase and preparing method and application thereof
A spore laccase and magnetic technology, applied in chemical instruments and methods, immobilized on or in inorganic carriers, and textile industry wastewater treatment, etc. The effect of preparation cost and high decolorization efficiency
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Embodiment 1
[0035] The production of embodiment 1 spore laccase
[0036] Cultivate the Bacillus pumilus W3 strain on the spore-forming medium at 37°C for 2 days, centrifuge the above culture at a centrifugal speed of 8000rpm for 10 minutes, collect the spores, then wash the spores once, and add lysozyme to the washed spores Keep the final concentration at 1mg / ml, incubate at 37°C for 2h, treat the culture with lysozyme, wash with 1M NaCl and 1M KCl successively, and collect spores by centrifugation at 8000rpm for 10min; wet weight the spores, and use Resuspend the spores in deionized water at a concentration of 100 mg / ml.
Embodiment 2
[0037] Example 2 Immobilization Scheme 1
[0038] Weigh 0.1g Fe 3 o 4 Magnetic nanoparticles (particle size is 20nm), added to 5ml 0.02M Na 2 HPO 4 -NaH 2 PO 4 Buffer (pH 7, containing 0.1M NaCl), add 0.5ml carbodiimide hydrochloride (4mg / ml), sonicate for 10min, then add 0.2ml spores (100mg / ml, wet weight), sonicate for 30min , and then placed at 200rpm, rotating and shaking at 37°C for 60min; the immobilized mixture was separated by magnetic adsorption, washed with buffer three times, and the supernatant and immobilized spores were collected and stored at 4°C. The recovery rate of enzyme activity was measured to be 92.97%.
Embodiment 3
[0039] Example 3 Immobilization scheme 2
[0040] Weigh 0.1g Fe 3 o 4 Magnetic nanoparticles (particle size is 20nm), added to 10ml 0.02M Na 2 HPO 4 -NaH 2 PO 4 Buffer (pH 7, containing 0.1M NaCl), add 0.5ml carbodiimide hydrochloride (4mg / ml), sonicate for 10min, then add 0.2ml spores (100mg / ml, wet weight), sonicate for 30min , and then placed at 200rpm, rotating and shaking at 37°C for 60min; the immobilized mixture was separated by magnetic adsorption, washed with buffer three times, and the supernatant and immobilized spores were collected and stored at 4°C. The recovery rate of enzyme activity was measured to be 90.12%.
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