Fungistatic paenibacillus sp. fermentation liquor extract
A technology of bacillus and fermentation liquid, applied in the field of microbiology, can solve problems such as teratogenicity, affecting food flavor, and lack of food, and achieve broad-spectrum and high-efficiency antibacterial effects, and the effect of inhibiting or killing pathogenic bacteria
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Embodiment 1
[0045] Example 1 The extraction of the antibacterial substance derived from Paenibacillus polymyxa (Paenibacillus polymyxa) BD3736
[0046] 1. Cultivation of BD3736 strain
[0047] The BD3736 strain preserved in 20% glycerol was inoculated on LB solid medium, and placed in an incubator at 30°C for 48 hours under atmospheric conditions. Then, a ring of single colony was scraped and placed in 15 mL of sterilized skim milk with a solid content of 10%, and incubated at a constant temperature of 30° C. and 180 r / min for 24 hours to obtain a BD3736 strain seed fermentation broth.
[0048] In a 500mL conical flask, 130mL of sterilized skim milk with a solids content of 10% was loaded, and the above-mentioned BD3736 strain seed fermentation liquid was inoculated according to the inoculation amount of 4% (v / v), and was shaken at a constant temperature of 30°C and 180r / min for 5 days. Obtain BD3736 strain fermentation broth.
[0049] 2. Preparation of BD3736 strain fermentation broth ...
Embodiment 2
[0051] The antibacterial spectrum of embodiment 2 BD3736 bacterial strain fermented liquid extract
[0052] 1. Preparation of indicator bacteria plate
[0053] The pathogenic bacteria preserved in 20% glycerol were respectively transferred to BHI solid medium, and cultured at 30°C for 48h. Then a ring of single colony was scraped and transferred to liquid BHI, and cultivated at 30°C and 180r / min for 24h. Centrifuge at 12000rpm for 5min to collect the bacterial cells, dilute with sterile water to make bacterial suspension. Use a hemocytometer to count and control the final bacterial concentration to 10 6 cfu / mL.
[0054] After the BHI solid medium is cooled to 55°C, mix it with the prepared indicator bacteria suspension and pour it into a plate, and pour about 20mL of medium into each petri dish. After solidification and sufficient drying, the indicator bacteria plate can be obtained.
[0055] 2. Antibacterial spectrum of BD3736 strain fermentation broth extract
[0056] ...
Embodiment 3
[0058] Example 3 Purification of BD3736 strain fermentation broth extract
[0059] (1) The crude product of the fermentation liquid extract of the BD3736 strain described in Example 1 was dialyzed overnight at 4° C. using dialysis bags with cut-off values of 3500 Da and 1000 Da respectively, and the samples in the bags were collected. Freeze-dry the dialysate to obtain freeze-dried powder.
[0060] (2) The lyophilized powder obtained in step (1) was dissolved in ultrapure water to a mass concentration of 200 mg / mL, and the sample volume was 10 mL, and separated by Sephadex G-15 (column volume about 800 mL) gel column chromatography. The eluent was ultrapure water with pH 7.0, and the flow rate was 3 mL / min. Collect one tube every 4min, about 12mL per tube. It was detected at 280nm with a protein UV detector. See the experimental results figure 2 .
[0061] The samples collected above were evaporated to dryness of the aqueous phase. Determination of the diameter of the...
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