Lumbricus polypeptide having anticoagulant and thrombolytic effects, and enzymatic hydrolysis preparation method and application thereof
An anticoagulant and thrombin technology, applied in the preparation method, application, food preparation and other directions of peptides, can solve the problems of easy to cause immunogenicity, low activity, reduced activity, etc., to improve bioavailability, structure Easy-to-understand, highly controllable effects
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Embodiment 1
[0034] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 20g: 100mL, adjust the pH to 9.0, stir evenly at a speed of 200r / min for 3h, take the supernatant, and then adjust the pH to 4, and finally obtain protein precipitation. The pH was adjusted to 7.0 and dried to obtain ground dragon protein. Add groundworm protein to water to dissolve, the ratio of protein to water is 20g: 100mL, control the pH value at 10, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to the weight of groundworm protein is 0.6 mL: 100g, the reaction temperature is 50°C, the stirring speed is 200r / min, and the reaction time is 1h. Boil for 10min; then adjust the pH value to 7.5, add trypsin (Sigma company, the U.S.), the ratio of enzyme to ground dragon protein weight is 0.5g: 100g, the reaction temperature is 50 ° C, the stirring speed is 200r / min, the reaction time 1h. Boil for 10min. Pass the enzymatic solution through a DEAE ...
Embodiment 2
[0036] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 25g:100mL, adjust the pH to 9.5, stir evenly at a speed of 250r / min for 4h, take the supernatant, and then adjust the pH to 4.5, and finally obtain protein precipitation, The pH was adjusted to 7.0 and dried to obtain ground dragon protein. Add ground dragon protein to water and dissolve, the ratio of protein to water is 25g: 100mL, control the pH value at 9.5, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to ground dragon protein weight is 0.8 mL: 100g, the reaction temperature is 60°C, the stirring speed is 250r / min, and the reaction time is 1.5h. Boil for 10 minutes; then adjust the pH value to 8.0, add trypsin (Sigma company, the United States), the weight ratio of enzyme and earthworm protein is 1g:100g, the reaction temperature is 50°C, the stirring speed is 200r / min, and the reaction time is 1h . Boil for 10min. Pass the enzymatic soluti...
Embodiment 3
[0038] Take 500g of clean Dilong dry powder, the ratio of dry powder to water is 30g:100mL, adjust the pH to 10, stir evenly at a speed of 300r / min for 5h, take the supernatant, then adjust the pH to 5.0, and finally obtain protein precipitation, The pH was adjusted to 7.0 and dried to obtain ground dragon protein. Add groundworm protein to water to dissolve, the ratio of protein to water is 30g: 100mL, control the pH value at 8.5, then add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the ratio of enzyme volume to the weight of groundworm protein is 1.0 mL: 100g, the reaction temperature is 60°C, the stirring speed is 300r / min, and the reaction time is 1h. Boil for 10min; then adjust the pH value to 8.5, add trypsin (Sigma company, the U.S.), the ratio of enzyme to earthworm protein weight is 1.5g: 100g, the reaction temperature is 45 ° C, the stirring speed is 300r / min, the reaction time 1h. Boil for 10min. Pass the enzymatic solution through a DEAE sepharo...
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