A kind of nattokinase and natto antibacterial peptide mixed preparation and its preparation method and application
A technology of nattokinase and preparation, which is applied in the field of biomedicine, can solve the problem of lack of comprehensive utilization of natto extract, and achieve the effect of inhibiting bacteria
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[0024] The invention provides a kind of preparation method of nattokinase and natto antimicrobial peptide mixed preparation, comprising the following steps:
[0025] 1) mixing natto with water, grinding, and filtering to obtain the grinding filtrate;
[0026] 2) Add solid ammonium sulfate to the ground filtrate in step 1), let stand, take the first supernatant, sterilize, and obtain natto antimicrobial peptide solution;
[0027] 3) mixing the natto with the phosphate buffer solution, grinding, mixing the obtained ground material with the phosphate buffer solution, extracting, centrifuging, and taking the second supernatant;
[0028] 4) adding an ammonium sulfate aqueous solution to the second supernatant in step 3) for mixing, leaving it to stand, centrifuging, and taking the precipitate to obtain nattokinase;
[0029] 5) mixing the natto antimicrobial peptide solution described in step 2) with the nattokinase described in step 4), filtering and sterilizing, and adjusting the...
Embodiment 1
[0047] Embodiment 1 A kind of preparation method of nattokinase and natto antimicrobial peptide mixed preparation
[0048] 1. Add 500g of commercially available natto to 500mL of distilled water, fully grind it with a tissue grinder, filter it with gauze, and obtain the grinding filtrate and filter residue;
[0049] 2. According to the filter residue: distilled water = 1:1, add distilled water to the filter residue, repeat the leaching twice under the condition of 4°C, and the time of each leaching is 1h, and obtain the leaching filtrate;
[0050] 3. Combine the grinding filtrate and leaching filtrate, add 50g of solid ammonium sulfate, stand at 4°C for 12h, collect the first supernatant; add 50g of solid ammonium sulfate to the first supernatant again, stand at 4°C for 12h, collect The supernatant was autoclaved at 121°C for 20 minutes, and filtered to obtain the natto antimicrobial peptide solution;
[0051] 4. Add 500 g of commercially available natto to 500 mL of pre-cool...
Embodiment 2
[0055] Embodiment 2 A kind of preparation method of nattokinase and natto antimicrobial peptide mixed preparation
[0056] 1. Add 1500g of commercially available natto to 500mL of distilled water, fully grind with a tissue grinder, filter with gauze, and obtain the grinding filtrate and filter residue;
[0057] 2. According to the filter residue: distilled water = 3:1, add distilled water to the filter residue, repeat the leaching twice under the condition of 4°C, and the time of each leaching is 1h, and obtain the leaching filtrate;
[0058] 3. Combine the grinding filtrate and leaching filtrate, add 80g solid ammonium sulfate, let stand for 15h, collect the first supernatant; add 80g solid ammonium sulfate again in the first supernatant, let stand for 15h, collect the supernatant, Autoclave at 121°C for 20 minutes, and filter to obtain the natto antimicrobial peptide solution;
[0059] 4. Add 1500 g of commercially available natto to 500 mL of pre-cooled phosphate buffer so...
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