Method for separating antibacterial proteins from bacillus pumilus E14
A technology of Bacillus pumilus and E14, which is applied to the preparation method of peptides, methods based on microorganisms, chemical instruments and methods, etc., can solve the problems of large losses and high frequency of disease occurrence, and achieve the effect of convenient operation and simple and reasonable design
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Embodiment 1
[0030] Embodiment 1, a kind of method for isolating antibacterial protein from Bacillus pumilus E14, its steps are as follows:
[0031] (1) Fermentation production: Bacillus pumilus E14 ( Bacillus pumilus ) Inoculate the Bacillus pumilus E14 seed liquid into 2216E liquid medium, the inoculum amount is 5%, 180rpm, 28°C for 24h; obtain the fermentation liquid;
[0032] (2) Preparation of supernatant: ferment the bacteria liquid at 1200rpm, centrifuge at 4°C for 30min, centrifuge the supernatant again, repeat 2-3 times; obtain the supernatant;
[0033] (3) Ammonium sulfate precipitation: Dry and grind the ammonium sulfate, weigh different aliquots respectively according to the preparation table of saturated ammonium sulfate solution, add to the supernatant while stirring on ice, and obtain ammonium sulfate with a mass volume percentage of 40% solution, then stand on ice for 20 min, centrifuge at 4°C, 8000rpm for 5 min; take the supernatant, continue to add ammonium sulfate while...
Embodiment 2
[0047] Embodiment 2, with reference to figure 1 -5. Experiment on the method of isolating antibacterial proteins from Bacillus pumilus E14:
[0048] 1. Experimental method
[0049] 1.1 Detection of Bacillus pumilus E14 antagonistic activity
[0050] Bacillus pumilus E14 is preserved in CGMCC, General Microorganism Center of China Committee for Culture Collection of Microorganisms, with preservation number CGMCC No.6682.
[0051] In this study, the zone of inhibition method of filter paper was used. Inoculate Bacillus pumilus E14 in 2216E liquid medium, culture at 28°C for 20 h, draw 0.1 mL of culture solution and spread it evenly on the 2216E solid medium plate. Place the sterile filter paper in a petri dish, then pipette 50 μL of the antagonistic bacteria to be tested dropwise onto the filter paper, incubate at 28°C, observe the inhibition zone and measure the size of the inhibition zone after 16 hours.
[0052] 1.2 Determination of the location of the active substance o...
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