Ocean cephalopod skin glue collagen and preparation method thereof
A collagen and cephalopod technology, applied in the field of marine cephalopod type I collagen products and their preparation, can solve problems such as no patent reports on marine cephalopod original protein, avoid new pollution, high pollution, etc. Safety and the effect of increasing added value
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Embodiment 1
[0032] 1. Raw material pretreatment
[0033] Fresh squid skin, washed with distilled water;
[0034] 2. Use eluate I to remove water-soluble impurities
[0035] Pound the cleaned squid skin with a tissue masher, add 10 times the amount of eluate I to homogenize, centrifuge (5000×g, 15min) to get the precipitate, then add eluate I to repeat the treatment twice;
[0036] 3. Use eluent II to remove salt-soluble impurities
[0037] Add 10 times the amount of eluate II to the precipitate obtained through the above steps, homogenize, stir and extract for 24 hours, centrifuge to collect the precipitate, and wash the precipitate twice with clean water;
[0038] 3. Use the extract to extract collagen
[0039] Add 10 times the amount of extract to the above precipitation, stir and extract for 48 hours, centrifuge to take the supernatant to obtain the crude collagen extract;
[0040] 4. Salting out of collagen
[0041] Add solid NaCl to the crude collagen extract to make the final c...
Embodiment 2
[0047] Wash the fresh or thawed cuttlefish skin with water, cut into pieces, homogenize, add 5 times the amount of eluate I for post-treatment, centrifuge to remove the supernatant, add 10 times the amount of eluate II to the precipitate, Stir and extract for 16h. Take the precipitate by centrifugation, wash the precipitate with clean water, and then use 15 times the amount of the extract, stir and extract for 56 hours to obtain a crude collagen extract. Add solid NaCl to make the final concentration reach 2.5M, centrifuge to discard the supernatant, dissolve the precipitate with 0.5M acetic acid, and perform dialysis, use 0.5M acetic acid as the dialyzed external fluid until the dialysate is free of Cl - So far, after being freeze-dried, squid skin collagen product is obtained, its molecular weight is 330KD, intrinsic viscosity is 18dL / g, denaturation temperature is 30°C, and every 1000 amino acid residues contains glycine, proline and Hydroxyproline is 330, 90, 70 respectiv...
Embodiment 3
[0049] Thaw the frozen squid skin in the storage room of the refrigerator, clean it with deionized water, then add 10 times the amount of eluate I, and homogenize it in a tissue masher. Centrifuge (2000×g, 15min), remove the supernatant, and stir and leach the precipitate with 15 times the amount of eluate II for 48h. Take the precipitate by centrifugation, wash the precipitate with deionized water for 3 times, add 20 times the amount of extract containing 1% pepsin, stir and extract for 72 hours, and centrifuge to get the supernatant, which is the crude collagen extract. Add solid KCl to make the final concentration up to 2.5M, centrifuge to remove the supernatant, dissolve the precipitate with 0.5M acetic acid, and perform dialysis treatment. The external fluid of the dialysis is 0.5M acetic acid. A squid skin collagen product is obtained after drying. After SDS-PAGE and amino acid analysis, it was determined to be type I collagen, and the molecular type was: [α1(I)] 2 The...
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