Silver carp dpp-iv inhibitory polypeptide and its application
A DPP-IV, silver carp technology, applied in the field of medicine, can solve problems such as no relevant research on DPP-IV inhibitory active peptides, and achieve good DPP-IV inhibitory activity, beneficial to human body utilization, and high DPP-IV inhibitory activity. Effect
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Method used
Examples
Embodiment 1
[0041] Embodiment 1: the preparation of silver carp meat protein solution
[0042] After the silver carp is killed, remove the scales and viscera, wash and collect the meat, select the white meat on the back of the silver carp, cut it into small pieces and use a grinder to stir it into minced meat; then mix the minced silver carp with deionized water, adjust The protein concentration is 5%, and the fish meat turbid liquid with adjusted protein concentration is placed in a 95°C water bath for 10 minutes to kill the enzyme, so as to eliminate the endogenous protease in the fish meat; after cooling, carry out homogenization treatment to obtain silver carp fish meat protein solution, freeze-dried for future use or directly proceed to the enzymatic hydrolysis reaction in the next step.
Embodiment 2
[0043] Embodiment 2: Preparation of freshwater fish silver carp polypeptide
[0044] The first step of enzymatic hydrolysis reaction: take a certain amount of fish protein powder and dissolve it in distilled water (5%, w / w). Place the solution in a constant temperature water bath at 37°C. When the temperature of the solution rises to 37°C, adjust its pH value to 2.0 with 1mol / L HCl, and add pepsin to the solution to start the first step of enzymatic hydrolysis reaction. The weight ratio of the substance, that is, E:S is 1%, and after hydrolysis at 37° C. for 1 hour, the hydrolyzate of pepsin is obtained;
[0045] The second step of enzymatic hydrolysis reaction: use 0.9mol / L NaHCO 3 Adjust the pH value of the hydrolyzate of pepsin to 5.3, then adjust its pH value to 7.5 with 1mol / L NaOH, then add trypsin, the weight ratio of enzyme and substrate, that is, E:S is 2%, start the second step Enzymolysis reaction, after 2 hours of reaction at 37°C, the enzymatic hydrolyzate of tr...
experiment example 3
[0048] Experimental Example 3: Optimization of Enzymolysis Reaction Conditions
[0049] Using the method of Example 2, the enzymatic hydrolyzate and reaction conditions of the enzymatic hydrolysis reaction were optimized, and the DPP-IV inhibitory activity in the final product was detected. The results are shown in Table 1:
[0050] Table 1: Enzymolysis reaction conditions and the degree of hydrolysis and DPP-Ⅳ inhibitory activity during the enzymolysis process
[0051] Protease type
Substrate concentration
pH
temperature
Enzyme / substrate ratio
Enzymolysis time
degree of hydrolysis
DPP-Ⅳ inhibition rate
Before enzymatic digestion
/
/
/
/
0h
0
7.74％
Pepsin
5％
2.0
37℃
1％
1h
7.88％
40.57％
trypsin
5％
8.0
37℃
2％
3h
20.26％
47.98％
Dispase
5％
7.5
45℃
3％
3.5h
22.71％
49.34％
Dispase
5％
7.5
45℃
...
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