Method for extracting catechol 2,3-dioxygenase from microalgae and purifying catechol 2,3-dioxygenase
A catechol and dioxygenase technology, applied in biochemical equipment and methods, redox enzymes, enzymes, etc., to achieve the effect of simple operation method, easy operation conditions and high purity
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Embodiment 1
[0025] (1) Microbiological identification and separation of water samples collected from the scene of the oil spill accident, from which chlorella was screened out, and the chlorella was enriched and cultivated: the medium was boiled with seawater for 15 minutes, and nutrient salts were added after suction filtration , the concentration of each component in the nutrient salt is NH 4 Cl 0.2g / L, K 2 HPO 4 0.05g / L and ferric citrate 0.01g / L, after 15 days, the algae liquid was obtained;
[0026] (2) Suction filter the algae solution obtained in step (1), wrap the filter membrane enriched with algae with tinfoil, place it in liquid nitrogen to cool for 30 seconds, take it out, add 4g of quartz sand in a mortar at 4°C Grinding at 1.2mmol / L pH 7.0 phosphate buffer solution to wash out the algae liquid, centrifuging at 10000r / min for 15min in a centrifuge, and taking the supernatant; the precipitated part was washed twice with phosphate buffer solution, combined and centrifuged th...
Embodiment 2
[0030] (1) Microbiological identification and separation of water samples collected from the scene of the oil spill accident, from which chlorella was screened out, and the chlorella was enriched and cultivated: the medium was boiled with seawater for 10 minutes, and nutrient salt was added after suction filtration , the concentration of each component in the nutrient salt is NH 4 Cl 0.1g / L, K 2 HPO 4 0.1g / L and ferric citrate 0.005g / L, after 10 days the algae liquid was obtained;
[0031] (2) Suction filter the algae solution obtained in step (1), wrap the filter membrane enriched with algae with tinfoil, place it in liquid nitrogen to cool for 90 seconds, take it out, add 3g of quartz sand in a mortar at 0°C Grinding at 1.5mmol / L, pH 7.5 phosphate buffer solution to wash out the algae liquid, centrifuged at 10000r / min for 15min in a centrifuge, and took the supernatant; the precipitated part was washed twice with phosphate buffer solution and combined for 3 The supernata...
Embodiment 3
[0035] (1) Microbiological identification and separation of water samples collected from the scene of the oil spill accident, from which chlorella was screened out, and the chlorella was enriched and cultivated: the medium was boiled with seawater for 20 minutes, and nutrient salts were added after suction filtration , the concentration of each component in the nutrient salt is NH 4Cl 0.3g / L, K 2 HPO 4 0.075g / L and ferric citrate 0.02g / L, after 7 days the algae liquid was obtained;
[0036] (2) Suction filter the algae solution obtained in step (1), wrap the filter membrane enriched with algae with tinfoil, place it in liquid nitrogen to cool for 60 seconds, take it out, add 5g of quartz sand in a mortar at 2°C Grind at 1.0mmol / L, pH 7.5 phosphate buffer solution to wash out the algae liquid, centrifuge at 10,000r / min for 15min, and take the supernatant; wash the precipitated part twice with the above phosphate buffer solution, and combine The supernatant of 3 centrifugati...
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