A bacteriostatic Paenibacillus fermentation broth extract
A technology of bacillus and fermentation broth, applied in the field of microorganisms, can solve the problems of teratogenicity, lack, affecting food flavor, etc., and achieve the effect of inhibiting or killing pathogenic bacteria, broad-spectrum and high-efficiency bacteriostatic effect
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Embodiment 1
[0044] Example 1 The extraction of the antibacterial substance derived from Paenibacillus polymyxa (Paenibacillus polymyxa) BD3736
[0045] 1. Cultivation of BD3736 strain
[0046] 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.
[0047] 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.
[0048] 2. Preparation of BD3736 strain fermentation broth ...
Embodiment 2
[0050] The antibacterial spectrum of embodiment 2 BD3736 bacterial strain fermented liquid extract
[0051] 1. Preparation of indicator bacteria plate
[0052] 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.
[0053] 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.
[0054] 2. Antibacterial spectrum of BD3736 strain fermentation broth extract
[0055] ...
Embodiment 3
[0057] Example 3 Purification of BD3736 strain fermentation broth extract
[0058] (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.
[0059] (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 .
[0060] The samples collected above were evaporated to dryness of the aqueous phase. Determination of the diameter of the...
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