A kind of antihypertensive active tripeptide and its preparation method and application
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Embodiment 1
[0016] Example 1 Preparation of Alcalase Enzyme Hydrolysis Active Peptide of Casein
[0017] Referring to CN 200710042207.5, the specific steps are:
[0018] 1. Enzyme reaction using enzyme reactor:
[0019] (1) Casein dissolution
[0020] Add 1000ml of water into the reactor, heat up to the optimum temperature of about 55°C, weigh 150g of casein (Shanghai Chemical Reagent Company), slowly and gradually put it into the water under magnetic stirring, adjust the pH to about 8.5 with 2mol / L NaOH, Under the condition of 55°C, all the casein was dissolved, so that the concentration of casein in the reactor was 15% (w / v, g / ml).
[0021] (2) Enzymolysis
[0022] According to the ratio of enzyme activity unit / substrate weight of 0.072AU / g casein, add 4.5g food-grade protease Alcalase (Novozymes Biotechnology Co., Ltd., 2.4L, enzyme activity 2.4AU / g), and maintain the reaction temperature at 55 ℃, and 2mol / L NaOH was added dropwise through the ZDJ-4A automatic potentiometric titrat...
Embodiment 2
[0029] Embodiment 2 Preparation of antihypertensive active tripeptide of the present invention
[0030] 1. Size exclusion chromatography separation process
[0031] Sephadex G-15 was packed into a 1.6 x 100 cm column. Get the Alcalase enzymatic hydrolysis active peptide sample of casein of embodiment 1, dissolve with pH3.6, 0.02moL / L acetic acid buffer solution and be prepared into the solution 15mL of certain concentration 10mg / mL and put on the column, and with the same buffer solution at 42ml / h The flow rate is eluted, and at the same time, the wavelength (λ) is 220nm for detection with an ultraviolet detector, such as figure 2 As shown, a total of 6 peaks from peaks I to VI were separated, and the components of peak V were collected and freeze-dried.
[0032] 2. High performance liquid chromatography purification process
[0033] HPLC system: Waters600E (Waters company);
[0034] Analysis parameters:
[0035] Column: μBondapak C 18 3.9*300mm I.D. (Waters Company); ...
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