Preparation method and application of a kind of bacillus pumilus antibacterial active substance
A technology of Bacillus pumilus and antibacterial active substances, applied in the field of microbial technology and application, can solve problems such as research on active substances of Bacillus pumilus, and achieve the effects of stable product, short production cycle and high content
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Embodiment 1
[0067] A strain of Bacillus pumilus BP, which was preserved in the General Microbiology Center of China Committee for Microbial Culture Collection (CGMCC for short) on March 21, 2019, address: Courtyard 3, No. 1, Beichen West Road, Chaoyang District, Beijing No., Institute of Microbiology, Chinese Academy of Sciences), the deposit number is CGMCC NO.17424.
[0068] The biological characteristics of the bacterial strain: it is of the genus Bacillus, the thalline is thin rod-shaped, and Gram-positive. It grows transparent on the LB slope and can produce antibacterial active ingredients.
Embodiment 2
[0070] The culture method of bacillus pumilus short (Bacillus pumilus) BP, the steps are as follows:
[0071] (1) Inoculate the above-mentioned Bacillus pumilus into the seed culture medium, and culture it for 16 hours under the conditions of 37° C. and 180 rpm to obtain the seed liquid;
[0072] Described seed culture medium is LB broth culture medium, and every liter of component is as follows:
[0073] 10 grams of peptone, 10 grams of sodium chloride, 5 grams of yeast extract, and water to 1000 ml;
[0074] (2) Inoculate the seed solution prepared in step (1) into the fermentation medium at a ratio of 3% by volume, and cultivate it at 30° C. at 180 rpm for 48 hours to obtain a fermentation solution;
[0075] Described fermentation medium, every liter of component is as follows:
[0076] Glucose 30.0g, K 2 HPO 4 ×3H 2 O 7.0g, KH 2 PO 4 3.0g, (NH 4 ) 2 SO 4 1.5g, sodium citrate 3H 2 O0.5g, MgSO 4 ·7H 2 O 0.1g, pH7.0~7.2, add water to make up to 1L.
Embodiment 3
[0078] The preparation method of the antibacterial active substance derived from bacillus pumilus (Bacillus pumilus) BP, the steps are as follows:
[0079] (i) The fermented liquid prepared in Example 2 was centrifuged at 3500 rpm for 35 minutes, the supernatant was taken, and concentrated to 8 times the concentration of the original solution by rotary evaporation to obtain a concentrated solution;
[0080] (ii) The concentrated solution obtained in step (ii) is extracted with ethyl acetate, the volume ratio of the concentrated solution to ethyl acetate is 1:1, the extraction phase is taken, extracted 4 times, the combined extraction phases are concentrated, and then subjected to 200 Objective Silica gel column chromatography separation, different gradient elution with different solutions of petroleum ether, ethyl acetate and methanol respectively, collecting petroleum ether / ethyl acetate=50:50 eluate with antibacterial activity, concentrating, blowing and drying with nitrogen,...
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