A kind of bacillus subtilis bacterial strain and application thereof
The technology of Bacillus subtilis and strain is applied to the Bacillus subtilis strain and its application field to achieve the effects of low nutrient loss rate, high hardness and broad application prospects
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Embodiment 1
[0026] The present embodiment provides a method for preparing a high density Bacillus subtilis fermentation broth comprising the steps of:
[0027] 1) Seed strain was prepared: Bacillus subtilis NA plate medium on activation, a single colony was streaked on NA medium, the sealant 33 ℃ upside down incubator 24h, to give the activated strains, strains with shift the activated rings were transferred to strain in LB medium, at 41.5 ℃, 240r conditions / min, and 37h culture, seed bacteria solution obtained were stored at 1.5 ℃;
[0028] Preparation 2) high density fermentation liquid: Step 1) Preparation of seed bacteria was good, at 4.5% (V / V) was inoculated in the flask in a high density medium at a temperature of 41.5 ℃, 240r under / min condition, 36h culture, Bacillus subtilis CGMCC NO.16569 achieve viable count of 3.27 × 10 11 CFU / ml. Obtained bacterial suspension, the bacterial suspension after centrifugation was 10000r / min and the supernatant sterilized by filtration with...
Embodiment 2
[0030] The present embodiment provides a method for preparing a high density Bacillus subtilis fermentation broth comprising the steps of:
[0031] 1) Seed strain was prepared: Bacillus subtilis NA plate medium on activation, a single colony was streaked on NA medium, the sealant 33 ℃ upside down incubator 24h, to give the activated strains, strains with shift the activated rings were transferred to strain in LB medium, at 33 ℃, 180r conditions / min culturing 24h, resulting seed bacteria solution were stored at -1 deg.] C;
[0032] Preparation 2) high density fermentation liquid: Step 1) Preparation of seed bacteria was good, at 3% (V / V) was inoculated in the flask in a high density medium at a temperature of 33 ℃, 180r under / min conditions, culture 24h, Bacillus subtilis CGMCC NO.16569 achieve viable count of 3.41 × 10 11 CFU / ml. Obtained bacterial suspension, the bacterial suspension after centrifugation was 10000r / min and the supernatant sterilized by filtration with m...
Embodiment 3
[0034] The present embodiment provides a method for preparing a high density Bacillus subtilis fermentation broth comprising the steps of:
[0035] 1) Seed strain was prepared: Bacillus subtilis NA plate medium on activation, a single colony was streaked on NA medium, the sealant 33 ℃ upside down incubator 24h, to give the activated strains, strains with shift the activated rings were transferred to strain in LB medium, at 50 ℃, 300r conditions / min culturing 5Oh, resulting seed bacteria solution were stored at 4 ℃;
[0036] Preparation 2) high density fermentation liquid: Step 1) Preparation of seed bacteria was good, at 6% (V / V) was inoculated in the flask in a high density medium at a temperature of 50 ℃, 300r under / min conditions, culture 48h, Bacillus subtilis CGMCC NO.16569 achieve viable count of 3.60 × 10 11 CFU / ml. Obtained bacterial suspension, the bacterial suspension after centrifugation was 10000r / min and the supernatant sterilized by filtration with micropor...
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