A kind of biological fungicide and preparation method thereof
A kind of antifungal agent and biological technology, applied in the field of antibacterial, can solve the problems of expensive equipment, cumbersome operation, drug residue and other problems of physical control technology, and achieve the effect of fast reproduction speed, simple operation and high feasibility
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Embodiment 1
[0040] A kind of preparation method of biological fungicide of the present invention, comprises the steps:
[0041] (1) Preparation of B.amyloliquefaciens F1 fermentation broth
[0042]The B. amyloliquefaciens F1 strain was inoculated into a test tube containing 5 mL of nutrient broth at a 2% inoculation amount, and cultured on a shaker at 30° C. to obtain a B. amyloliquefaciens F1 seed solution. Aspirate B.amyloliquefaciens F1 seed liquid, add 2% inoculum amount to an Erlenmeyer flask with 40% fermentation medium loading, culture at 30°C on a shaking table for 48 hours, and obtain B.amyloliquefaciens F1 fermentation liquid.
[0043] (2) Preparation of B.amyloliquefaciens F1 crude antibacterial protein
[0044] First, take 1 L of the above-mentioned B. amyloliquefaciens F1 fermentation broth and perform ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 15 kDa to obtain an ultrafiltration concentrate. Then slowly add ammonium sulfate to the u...
Embodiment 2
[0049] A kind of preparation method of biological fungicide of the present invention, comprises the steps:
[0050] (1) Preparation of B.amyloliquefaciens F1 fermentation broth
[0051] The B. amyloliquefaciens F1 strain was inoculated into a test tube containing 5 mL of nutrient broth at a 2% inoculation amount, and cultured on a shaker at 30° C. to obtain a B. amyloliquefaciens F1 seed liquid. Aspirate B.amyloliquefaciens F1 seed liquid, add 2% inoculum amount to an Erlenmeyer flask with 20% fermentation medium loading, and culture on a shaker at 32°C for 72 hours to obtain B.amyloliquefaciens F1 fermentation liquid.
[0052] (2) Preparation of B.amyloliquefaciens F1 crude antibacterial protein
[0053] First, take 1 L of the above-mentioned B. amyloliquefaciens F1 fermentation broth and perform ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 15 kDa to obtain an ultrafiltration concentrate. Then ammonium sulfate was slowly added to the u...
Embodiment 3
[0060] A kind of preparation method of biological fungicide of the present invention, comprises the steps:
[0061] (1) Preparation of B.amyloliquefaciens F1 fermentation broth
[0062] The B. amyloliquefaciens F1 strain was inoculated into a test tube containing 5 mL of nutrient broth at a 2% inoculation amount, and cultured on a shaker at 30° C. to obtain a B. amyloliquefaciens F1 seed solution. Aspirate B.amyloliquefaciens F1 seed solution, add 2% inoculum amount to an Erlenmeyer flask with 30% loading capacity of fermentation medium, and culture on a shaker at 37°C for 70 hours to obtain B.amyloliquefaciens F1 fermentation solution.
[0063] (2) Preparation of B.amyloliquefaciens F1 crude antibacterial protein
[0064] First, take 1 L of the above-mentioned B. amyloliquefaciens F1 fermentation broth and perform ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 8 kDa to obtain an ultrafiltration concentrate. Then ammonium sulfate was slow...
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