Technique for preparing antimicrobial peptide by using Bacillus subtilis
A technology of Bacillus subtilis and process methods, applied in the direction of peptide preparation methods, microorganism-based methods, biochemical equipment and methods, etc., to achieve the effects of high safety, no drug resistance, and improved preparation efficiency
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Embodiment 1
[0043] A process for preparing antimicrobial peptides by using Bacillus subtilis according to the present invention The production strain is specifically Bacillus subtilis (Bacillus subtilis) BRT39. , the deposit number is CGMCC NO.5702.
[0044] The seed medium consists of: glucose 2g, peptone 1g, NaH 2 PO 4 2H 2 O0.2g, Na 2 HPO 4 2H 2 O0.4g, MgSO 4 ·7H 2 O0.05g, dH 2 O100mL, pH 7.5, sterilized at 121°C for 20min.
[0045] The shake flask fermentation medium consists of: soybean meal powder 1-2g, maltose 0.5g, glucose 0.7-1g, ammonium sulfate 0.3g, ammonium nitrate 0.2g, peptone 0.4-1g, beef extract 0.5g, NaCl 0.5g, KH 2 PO 4 0.1g, MgSO 4 ·7H 2 O0.04g, dH 2 O100mL, pH8.0, 121°C, sterilize for 20min.
[0046] The fermentation medium of the fermenter is composed of: soybean meal powder 1-2g, corn flour 0.5-1g, maltose 0.5g, soluble starch 0.5-0.7g, glucose 0.7-1g, ammonium sulfate 0.3g, ammonium nitrate 0.2g, peptone 0.4 -1g, beef extract 0.5g, NaCl 0.5g, KH 2 P...
Embodiment 2
[0048] A kind of antimicrobial peptide preparation process method, comprises the steps:
[0049] 1) Strain activation
[0050] The well-preserved slant strain CGMCC NO.5702 was inoculated on the slant medium and cultured at 35°C for 16 hours to activate the strain.
[0051] 2) Liquid seed culture
[0052] Insert a single colony of activated Bacillus subtilis into the sterilized seed medium, and culture it on a rotary shaker at 180r / min and 35°C for 16 hours, and the bacterial content of the seed fermentation liquid is 10 7 cfu / mL;
[0053] 3) Shake flask fermentation culture
[0054] The seed solution obtained in step 2) was inserted into the Erlenmeyer flask with a 5% inoculum amount, the 1000mL Erlenmeyer flask contained a fermentation liquid medium of 200mL, the Erlenmeyer flask stopper was a latex stopper, the fermentation temperature was 32°C, and the rotation speed was 160r / min, and the shaking table was cultivated for 60h;
[0055] 4) Fermentation culture in 15-lite...
Embodiment 3
[0066] A kind of antimicrobial peptide preparation process method, comprises the steps:
[0067] 1) Strain activation
[0068] The well-preserved slant strain CGMCC NO.5702 was inoculated on the slant medium and cultured at 36°C for 18 hours to activate the strain.
[0069] 2) Liquid seed culture
[0070] Insert a single colony of activated Bacillus subtilis into the sterilized seed culture medium, and cultivate it in a rotary shaker at 190r / min and 36°C for 18 hours, and the bacterial content of the seed fermentation liquid is 10 7 cfu / mL;
[0071] 3) Shake flask fermentation culture
[0072] The seed liquid obtained in step 2) was inserted into the Erlenmeyer flask with an inoculum amount of 8%, and the 1000mL Erlenmeyer flask contained a fermentation liquid medium of 200mL, and the Erlenmeyer flask stopper was made of latex, the fermentation temperature was 35°C, and the rotating speed was 180r / min, and the shaking table was cultivated for 61h;
[0073] 4) Fermentation cu...
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