Giant salamander active peptide and application
An active peptide, giant salamander technology, applied in the field of giant salamander active peptide, can solve problems such as single function, achieve broad application prospects, promote the proliferation of skin fibroblasts, and improve the effect of immunity
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Embodiment 1
[0026] Follow the steps below:
[0027] a. Homogenize giant salamander meat, add 1 volume of 0.01M phosphate buffer with pH 7 and a compound enzyme with a weight ratio of 0.1% to the homogenate, and enzymolyze it for 12 hours; the compound enzyme is marine alkaline Protease and papain are mixed according to the activity unit ratio of 7:3;
[0028] b. Take the supernatant by centrifugation, and separate it with a trypsin immobilized ultrafiltration membrane separator prepared by physical adsorption method with a molecular weight cut-off of 4000 Da or less, and take the liquid that passes through the ultrafiltration membrane;
[0029] c. Pass the Sephadex LH-20 molecular sieve chromatography column (15mm×1000mm), collect the chromatographic product in 1ml per tube, and take the 30th~36th tube of chromatographic product for freeze-drying, such as figure 1 Shown: the collected chromatographic product is the absorption peak of OD280nm (peak II);
[0030] d. The freeze-dried prod...
Embodiment 2
[0034] Follow the steps below:
[0035] a. Homogenize giant salamander meat, add 3 times the volume of 0.1M phosphate buffer with pH 8 and a compound enzyme with a weight ratio of 1% to the homogenate, and enzymolyze it for 48 hours; the compound enzyme is marine alkaline Protease and papain are mixed according to the activity unit ratio of 7:3;
[0036] b. Take the supernatant by centrifugation, and separate it with a trypsin immobilized ultrafiltration membrane separator prepared by physical adsorption method with a molecular weight cut-off of 4000 Da or less, and take the liquid that passes through the ultrafiltration membrane;
[0037] c. Pass the Sephadex LH-20 molecular sieve chromatography column (15mm×1000mm), collect the chromatographic product in 1ml per tube, and take the 30th~36th tube of chromatographic product for freeze-drying, such as figure 1 Shown: the collected chromatographic product is the absorption peak of OD280nm (peak II);
[0038] d. The freeze-dri...
Embodiment 3
[0042] Follow the steps below:
[0043] a. Homogenize giant salamander meat, add 2 times volume of 0.1M phosphate buffer with pH 8 and compound enzyme with a weight ratio of 0.5% to the homogenate, and hydrolyze for 34 hours; the compound enzyme is marine alkaline Protease and papain are mixed according to the activity unit ratio of 7:3;
[0044] b. Take the supernatant by centrifugation, and separate it with a trypsin immobilized ultrafiltration membrane separator prepared by physical adsorption method with a molecular weight cut-off of 4000 Da or less, and take the liquid that passes through the ultrafiltration membrane;
[0045]c. Pass the Sephadex LH-20 molecular sieve chromatography column (15mm×1000mm), collect the chromatography product in 1ml per tube, and take the 30th~36th tube of chromatography product for freeze-drying; figure 1 Shown: the collected chromatographic product is the absorption peak of OD280nm (peak II);
[0046] d. The freeze-dried product obtained...
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