Method for extracting superoxide dismutase from pig spleen
A technology of superoxide and dismutase, applied in biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of difficult large-scale production, small processing capacity, high production cost, etc., and achieve low production cost, simple operation and high efficiency The effect of recycling
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Embodiment 1
[0030] Take 10 kg of fresh pig spleen from the slaughterhouse, chop it up, add 50 liters of deionized water, refine it with a colloidal mill, centrifuge at 5000 rpm for 10 minutes, and filter with filter paper to obtain 60 liters of clear liquid, add 10.5 kg of solid ammonium sulfate (up to 30 % saturation), stirred for 6 hours below 25°C, filtered to remove the precipitate, added 11.88 kg of ammonium sulfate to the supernatant (up to 60% saturation), and filtered to obtain the precipitate. Add 20 liters of deionized water to dissolve the precipitate, add 10 ml of 2 moles of Tris-Cl (pH 8.0) to make the solution contain 10 millimoles of Tris-Cl (pH 8.0), and filter with filter paper to obtain a transparent liquid. Add 5 liters of copper-IDA agarose medium balanced with 10 millimolar Tris-Cl (pH8.0) to the transparent liquid, stir slowly (50 rpm) for 2 hours, stop stirring, and let it stand for 1 hour. Sieve and filter to obtain the medium with SOD adsorbed, wash the medium wit...
Embodiment 2
[0032] Take 100kg of frozen pig spleen, chop it up, add 500 liters of deionized water, grind it with colloid, remove coarse impurities by centrifugation in a horizontal centrifuge, and then use a small plate and frame filter to clarify and filter to obtain about 600 liters of clear liquid, add 105 kg of solid ammonium sulfate (up to 30% saturation), stirred for 6 hours below 25°C, filtered with a small plate and frame filter to remove the net precipitate, and then added 118.8 kg of ammonium sulfate (up to 60% saturation) to the supernatant, and used A small plate and frame filter was used to filter the precipitate. Add 200 liters of deionized water to dissolve the precipitate, add 100 ml of 2 moles of Tris-Cl (pH 8.0) to make the solution contain 10 millimoles of Tris-Cl (pH 8.0), and filter it with a small plate and frame filter to obtain Transparent liquid. Add 25 liters of copper-IDA agarose medium balanced with 10 mmol Tris-Cl (pH8.0) to the transparent liquid, stir slowl...
Embodiment 3
[0034] Take 500 kg of frozen pig spleen, chop it up, add 2.5 tons of deionized water, grind it with colloid, centrifuge at 5000 rpm for 10 minutes, filter it with a 4㎡ plate and frame filter, get about 3000 liters of clear liquid, add 528 kg of solid ammonium sulfate (up to 30% saturation), stir for 6 hours below 25°C, filter the precipitate with a 4㎡ plate and frame filter, add 594 kg of ammonium sulfate to the supernatant (up to 60% saturation), and use a 4㎡ plate and frame filter to filter the precipitate. Add 500 liters of deionized water to dissolve the precipitate, add 500 ml of 2 moles of Tris-Cl (pH 8.0) to make the solution contain 10 millimoles of Tris-Cl (pH 8.0), and filter it with a small plate and frame filter to obtain Transparent liquid. Add 50 liters of copper-IDA agarose medium balanced with 10 mmol Tris-Cl (pH8.0) to the transparent liquid, stir slowly (30 rpm) for 2 hours, stop stirring, and after standing for 1 hour, use 200 Filter through a mesh sieve t...
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