A kind of preparation method of goat milk casein source antimicrobial peptide
A technology of goat cheese and antimicrobial peptides, which is applied in the fields of peptide preparation methods, antibacterial drugs, chemical instruments and methods, etc. It can solve the problems of unclear primary and secondary structures of antimicrobial peptides, achieve excellent antibacterial properties and broad application prospects Effect
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[0031] 2) Preparation of casein hydrolyzate
[0032] Weigh an appropriate amount of goat milk casein crude product and place it in an enzymolysis tank, dissolve it with 0.1mol / L sodium hydroxide solution, and dilute to the specified volume with deionized water to obtain 30g / kg~120g / kg (optimized as 60g / kg~100g / kg), adjust the pH value to 5.0~7.0 (optimize 5.5~6.5), heat at 95℃ for 5min, cool to 50℃~70℃, add 3000U / g~7000U / g ( Optimize the papain of 4000U / g~6000U / g), add 2mol / L NaOH solution during the hydrolysis process to keep the pH value constant, after 0.5h~4.0h of enzymolysis (optimize 1.0h~2.0h), extinguish at 95℃ Enzyme for 10 minutes, cool to room temperature, centrifuge at 10,000×g for 15 minutes, take the supernatant and concentrate at 45-50°C, 0.09MPa to obtain goat cheese casein enzymatic hydrolysis concentrate (ie crude antimicrobial peptide).
[0033] 3) Desalting and separation and purification of antimicrobial peptides
[0034] After pretreatment of the macro...
Embodiment 1
[0055] (1) Take fresh goat milk and filter to remove impurities, centrifuge at 5000r / min at 4°C for 20 minutes to remove milk fat, adjust the pH value of the skim goat milk to 4.3 with 2mol / L HCl solution, and let it stand in a water bath at 40°C for 30 minutes to precipitate casein. Centrifuge at 4000r / min for 10min, discard the supernatant, wash the precipitate twice with acetic acid-sodium acetate buffer solution with pH4.3, and centrifuge at 4000r / min for 10min after each wash, the obtained precipitate is crude casein, Kjeldahl Determination of protein content of crude casein by nitrogen method.
[0056] (2) Weigh a certain amount of goat milk crude casein and place it in an enzymolysis tank, dissolve it with 0.1mol / L sodium hydroxide solution, and dilute it to the specified volume with deionized water to obtain 80g / kg goat cheese For protein solution, adjust the pH value to 6.0, heat at 95°C for 5 minutes, cool to 60°C, accurately add 5000U / g papain, and add 2mol / L NaOH s...
Embodiment 2
[0061] 1) Take fresh goat milk to filter and remove impurities, centrifuge at low temperature to degrease, adjust the pH value of the defatted goat milk to 4.3 with 2mol / L HCl solution, let stand in a water bath at 40°C for 30min to precipitate casein, centrifuge at 4000r / min for 10min, and discard the supernatant solution, the precipitate was washed twice with acetic acid-sodium acetate buffer solution of pH 4.3, and centrifuged at 4000r / min for 10min after each washing, the obtained precipitate was goat milk crude casein.
[0062] (2) Weigh a certain amount of goat milk crude casein and place it in an enzymolysis tank, dissolve it with 0.1mol / L sodium hydroxide solution, and dilute it to the specified volume with deionized water to obtain 70g / kg goat milk casein Milk casein solution, adjust pH value to 6.0, heat at 95°C for 5min, cool to 60°C, add 6000U / g papain accurately, add 2mol / L NaOH solution during hydrolysis, keep pH value at 6.0±0.1, 60 After 80 min of enzymatic hyd...
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