A kind of bacillus amyloliquefaciens and its preparation method and application
A technology of amyloliquefaciens and bacilli, applied in the direction of biochemical equipment and methods, applications, and methods based on microorganisms, to achieve the effects of promoting intestinal absorption, reducing pollution, and improving intestinal health
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Embodiment 1
[0025] Strain Isolation
[0026]Fresh feces from healthy pigs were collected from a natural pig farm in Huangpi, Hubei. The feces samples were enriched in nutrient broth (NB) medium for aerobic enrichment and turbidity. They were treated in a water bath at 80°C for 15 minutes, and then enriched and heated. Screen once, spread on plate after appropriate dilution, pick typical single colonies according to the colony shape (colonies are nearly round, 3-4.5mm, dull, central bulge, edge cog-toothed, off-white), and spores are screened out by microscopic examination bacilli.
[0027] The formula of NB medium (1L): beef extract 5g, peptone 10g, NaCl 5g, agar 15g.
[0028] Re-screening and identification
[0029] Screened against Staphylococcus aureus (G + ), Escherichia coli (G - ) and other pathogenic bacteria with good growth inhibition performance, through physiological and biochemical and 16s identification, a strain of Bacillus amyloliquefaciens was finally screened and name...
Embodiment 2
[0031] Example 2: Scale-up cultivation of Bacillus amyloliquefaciens H6
[0032] According to corn flour 15g / L, soybean meal 20g / L, yeast extract 10g / L, peptone 10g / L, CaCO 3 3g / L, NaCl5g / L, MgSO 4 2g / L, KH 2 PO 4 3g / L, polyether 2ml / L, and 65% (V / V) liquid content to prepare fermentation broth, sterilized by high-temperature steam at 121°C for 20min, then cooled to 40°C, inoculated with 1% seed liquid of 10h age, Cultivate at 32°C, control the stirring speed to maintain dissolved oxygen (DO value above 30%), and automatically add defoamer (silicone oil) online. Sampling every 2 hours to measure the OD value, fermented for 24 hours and placed in the tank, the plate count of Bacillus amyloliquefaciens H6 was 1.78×10 10 cfu / ml ( figure 2 ).
Embodiment 3
[0033] Embodiment 3: preparation of bacillus amyloliquefaciens bacteria powder
[0034] The bacteria liquid fermented in Example 2 is coarsely filtered through a vibrating sieve to remove large solids, and then concentrated to 1 / 6 of the original volume by microfiltration through a 0.2 μm ceramic membrane. The concentrated solution is directly pumped into the spray drying tower by a peristaltic pump. 180±3°C, outlet temperature 67-70°C, control the feed flow rate and heating switch, maintain temperature balance, calculate the bacterial powder output after receiving the material to be 40kg / m 3 Fermentation liquid, plate count bacterial powder bacterial count is 4.7×10 11 cfu / g.
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