A kind of bacillus licheniformis with high tetramethylpyrazine production and its isolation and cultivation method and application
A technology of Bacillus licheniformis and tetramethylpyrazine, which is applied to the field of Bacillus licheniformis with high yield of tetramethylpyrazine and its separation and culture, can solve the problem of long period of koji-making process, energy consumption and difficulty in tetramethylpyrazine Efficient accumulation of oxazine and other problems, to achieve far-reaching social and economic significance, the effect of improving production efficiency
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Embodiment 1
[0065] The present embodiment provides a kind of Bacillus licheniformis high-yielding tetramethylpyrazine, and the separation method comprises the following steps:
[0066] (1) Mix high-temperature daqu and sterile saline to obtain a bacterial suspension;
[0067] (2) Dilute the bacterial suspension obtained in step (1) and treat it at 80°C for 20 minutes and then cultivate it on beef extract peptone solid medium to obtain colonies; beef extract peptone solid medium includes beef extract 3g / L, peptone 10g / L, sodium chloride 5g / L, agar powder 20g / L, solvent is water; pH=7.2;
[0068] (3) pick the morphologically regular bacterium colonies in step (2) and carry out streak culture on the beef extract peptone solid medium to obtain a single pure-bred bacterium colony;
[0069] (4) Pick a single colony for liquid culture, prepare a seed solution, inoculate it on a bran solid medium and cultivate it at 37°C for 24 hours, measure the content of tetramethylpyrazine, and obtain potent...
Embodiment 2
[0071] The present embodiment provides a kind of Bacillus licheniformis high-yielding tetramethylpyrazine, and the separation method comprises the following steps:
[0072] (1) Mix high-temperature daqu and sterile saline to obtain a bacterial suspension;
[0073] (2) Dilute the bacterial suspension obtained in step (1) and treat it at 85°C for 15 minutes and then cultivate it on beef extract peptone solid medium to obtain colonies; beef extract peptone solid medium includes beef extract 2g / L, peptone 12g / L, sodium chloride 6g / L, agar powder 22g / L, solvent is water; pH=7.3;
[0074] (3) pick the morphologically regular bacterium colonies in step (2) and carry out streak culture on the beef extract peptone solid medium to obtain a single pure-bred bacterium colony;
[0075] (4) Pick a single colony for liquid culture, prepare a seed solution, inoculate it on a bran solid medium and cultivate it at 37°C for 24 hours, measure the content of tetramethylpyrazine, and obtain potent...
Embodiment 3
[0077] The Bacillus licheniformis that embodiment 1 screens obtains carries out the observation of bacterium colony form and individual form, the result is as follows figure 1 and figure 2 as shown ( figure 1 It is the colony form figure of the bacillus licheniformis of high-yielding tetramethylpyrazine of the present invention, figure 2 It is the individual morphology diagram of Bacillus licheniformis with high tetramethylpyrazine production in the present invention). Depend on figure 1 It can be seen that the colonies of Bacillus licheniformis with high tetramethylpyrazine production in the present invention are flat, with irregular edges, white, and rough and wrinkled surfaces. Depend on figure 2 It can be seen that: under an optical microscope, the individual shape of the bacillus licheniformis of the high-yield tetramethylpyrazine of the present invention is rod-shaped, and there is a single existence, and there are also pairs and chains. The characteristics match...
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