A kind of bacillus subtilis and microbial agent and their application in fermentation and decomposing
A technology of Bacillus subtilis and microbial inoculum, which is applied in the field of agricultural biology, can solve the problem that the efficiency of fermentation and decomposing needs to be improved, and achieve the effects of improving the efficiency of fermentation and decomposing and improving the efficiency.
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Embodiment 1
[0047] This example is used to illustrate the cultivation of Bacillus subtilis and the preparation of microbial inoculum of the present invention.
[0048] The slant strain of Bacillus subtilis (Bacillus subtilis) with the preservation number of CCTCC NO: M2013496 was placed on an LB plate (containing 10g / L of peptone, 5g / L of yeast extract, 5g / L of NaCl and 3g / L of agar sugar, pH value 7.4, the same below), and cultured at 37°C for 15 hours, picked a single colony from the plate, and inoculated it in 25mL of LB liquid medium (containing 10g / L peptone, 5g / L Yeast extract and 5g / L NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain primary seeds, and added 20mL of primary seeds to 500mL of LB liquid medium In 250r / min shaker at 37°C for 15 hours to obtain secondary seeds, 2L of secondary seeds were added to 50L of LB liquid medium, stirred at 250r / min at 37°C and 10m 3 / h ventilation for 15 hours to obtain a culture solu...
Embodiment 2
[0082] This example is used to illustrate the cultivation of Bacillus subtilis and Bacillus licheniformis of the present invention and the preparation of microbial inoculum.
[0083] The slant strain of Bacillus subtilis (Bacillus subtilis) with the preservation number of CCTCC NO: M2013496 was placed on an LB plate (containing 10g / L of peptone, 5g / L of yeast extract, 5g / L of NaCl and 3g / L of agar sugar, pH value 7.4, the same below), and cultured at 37°C for 15 hours, picked a single colony from the plate, and inoculated it in 25mL of LB liquid medium (containing 10g / L peptone, 5g / L Yeast extract and 5g / L NaCl, pH value 7.4, hereinafter the same), cultivated on a shaker at 250r / min at 37°C for 15 hours to obtain primary seeds, and added 20mL of primary seeds to 500mL of LB liquid medium , cultivated on a shaker at 250 r / min at 37°C for 15 hours to obtain secondary seeds of Bacillus subtilis.
[0084] The slant strain of Bacillus licheniformis (Bacillus licheniformis) with th...
Embodiment 3
[0088] The microbial inoculum was prepared according to the method of Example 2, except that 1.2 L of secondary seeds of Bacillus subtilis and 1.2 L of secondary seeds of Bacillus licheniformis were mixed, and then tertiary expansion culture was carried out.
[0089] In this embodiment, the CFU ratio of the Bacillus subtilis to the Bacillus licheniformis in the culture medium was determined to be 1:0.9 by using the nucleic acid quantification method based on 16S rDNA.
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Abstract
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