Preparation method of thymosin
A technology of thymosin and thymus, applied in the field of preparation of thymosin, can solve the problems of high molecular weight of products, residual chemical reagents, low extraction efficiency of small molecular polypeptides, etc., and achieve the effects of high purity, controllable molecular weight and easy operation.
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Embodiment 1
[0028] This embodiment provides a kind of preparation method of thymosin, specifically comprises the following steps:
[0029] Step a, select 1000g of fresh porcine thymus, and use a liquid nitrogen grinder to grind it with liquid nitrogen as a raw material;
[0030] Step b, add 5000mL of purified water to the raw material, stir evenly, after ultrasonication for 20min, heat up to 35°C, add 8g of Candida lipase and 10g of Bacillus subtilis protease, keep for 40min, then add 3g of lactic acid bacteria aminopeptidase, keep The temperature is 4h, and finally the temperature is raised to 90°C, and the enzyme is kept for 15min;
[0031] Step c, filter with horizontal gong centrifuge (centrifugal speed is 3400r / min) after step c, deenzyme, filtrate is degreased and purified with three-phase centrifuge again; Use a nanofiltration machine with a nanofiltration membrane pore size of 0.18nm to desalt and concentrate, and the concentrate is further purified with an ion filter, and then f...
Embodiment 2
[0034] This embodiment provides a kind of preparation method of thymosin, specifically comprises the following steps:
[0035] Step a, select 10 kg of fresh bovine thymus, and use a liquid nitrogen grinder to grind it with liquid nitrogen as a raw material;
[0036] Step b: Add 60L of purified water to the raw material, stir evenly, after ultrasonication for 25min, heat up to 40°C, add 50g of Candida lipase, 50g of Aspergillus niger lipase and 120g of Bacillus subtilisin, keep for 35min, then add 40g For lactic acid bacteria aminopeptidase, keep the temperature for 3.5 hours, and finally raise the temperature to 93°C, and keep it for 12 minutes to kill the enzyme;
[0037] Step c, filter with horizontal gong centrifuge (centrifugal speed is 3600r / min) after step c, deenzyme, and filtrate degreases and purifies with three-phase centrifuge again; Use a nanofiltration machine with a nanofiltration membrane pore size of 0.2nm to desalt and concentrate, and the concentrate is furthe...
Embodiment 3
[0040] This embodiment provides a kind of preparation method of thymosin, specifically comprises the following steps:
[0041] Step a, select 100kg of fresh sheep thymus, and use a liquid nitrogen grinder to grind it with liquid nitrogen as a raw material;
[0042] Step b, add 400L of purified water to the raw material, stir evenly, after ultrasonication for 30min, heat up to 45°C, add 1.2kg of Aspergillus niger lipase and 1.5kg of Bacillus subtilis protease, keep for 30min, then add 0.5kg of lactic acid bacteria aminopeptidase, Keep the temperature for 3 hours, and finally raise the temperature to 95°C, and keep it for 10 minutes to inactivate the enzyme;
[0043] Step c, filter with horizontal gong centrifuge (centrifugal speed is 3800r / min) after step c, deenzyme, filtrate is degreased and purified with three-phase centrifuge again; Use a nanofiltration machine with a nanofiltration membrane pore size of 0.22nm to desalt and concentrate, and the concentrate is further puri...
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