A kind of preparation method of placental polypeptide
A placental polypeptide, placenta technology, applied in the directions of hydrolase, fermentation, etc., can solve the problem of low yield of polypeptide, and achieve the effects of simple preparation method, low production cost and less equipment investment
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Embodiment 1
[0021] The placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded and added to normal saline at a mass ratio of 1:2 for tissue homogenization; the resulting tissue homogenate is mixed with a compound enzyme at a mass ratio of 50:1, and the The complex enzyme is composed of trypsin and papain with a mass ratio of 1:1. It is hydrolyzed for 4 hours at 30°C and pH 5, and the centrifuge is collected by centrifugation at 5000r / min. The collected centrifugate is used as the pre-extraction aqueous phase. Hexanol and petroleum ether with a volume ratio of 1:3 are the organic phase, wherein the volume ratio of the pre-extracted organic phase and the aqueous phase is 2:1, and AOT and Span-60 with a mass ratio of 2:1 are used as a mixed surfactant. Extract at 40°C for 15 minutes, use 2mol / L KCl solution as the stripping water phase, wherein the volume ratio of the stripped organic phase to the KCl solution water phase is 2:1, strip at 35°C for 30 minutes, and the e...
Embodiment 2
[0023] The placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded and added to normal saline at a mass ratio of 1:2.5 for tissue homogenization; the resulting tissue homogenate is mixed with a compound enzyme at a mass ratio of 55:1, and the The compound enzyme is composed of five kinds of trypsin, pepsin, papain, bromelain and carboxypeptidase of the same quality. It is hydrolyzed at 35°C and pH 6 for 4.5h, centrifuged at 5000r / min to collect the centrifugate, and the collected The centrifuge liquid is used as the pre-extraction aqueous phase, and the hexanol and sherwood oil of 1:4 volume ratio are organic phases, wherein the organic phase and the aqueous phase volume ratio of pre-extraction are 2.5:1, with the AOT of 2.5:1 mass ratio and division Pan-60 is a mixed surfactant, which is extracted at 42°C for 16 minutes, and 2mol / L KCl solution is used as the stripping water phase, where the volume ratio of the stripping organic phase to the KCl solutio...
Embodiment 3
[0025] After the placenta without hepatitis B, hepatitis C, AIDS, and syphilis virus is shredded, add physiological saline in a mass ratio of 1:3 to carry out tissue homogenization; the resulting tissue homogenate is mixed with papain in a mass ratio of 60:1, and the ℃, pH 9 conditions of enzymatic hydrolysis for 5h, centrifugation at 5000r / min to collect the centrifugate, the collected centrifugate as the pre-extraction aqueous phase, with 1:5 volume ratio of hexanol and petroleum ether as the organic phase, of which the pre-extracted The volume ratio of the organic phase to the aqueous phase is 3:1, with AOT and Span-60 in a mass ratio of 3:1 as the mixed surfactant, extract at 45°C for 20min, and use 2mol / L KCl solution as the back extraction aqueous phase , where the volume ratio of the back-extracted organic phase to the aqueous phase of the KCl solution is 3:1, back-extracted at 40°C for 40 minutes, and the crude extract of placental polypeptide and protease is obtained a...
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