Bacillus licheniformis and application thereof in promoting soy sauce fermentation and aroma enhancement
A technology of Bacillus licheniformis and soy sauce, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., to achieve the effect of promoting ability and decomposition ability
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Embodiment 1
[0039] Isolation, cultivation and molecular identification of Bacillus licheniformis Mut-148:
[0040] The wild Bacillus genus XS-4 screened from the sauce fermented grains in the laboratory in the early stage (the colony morphology on the NB agar medium is as follows: Figure 6 Shown) to carry out atmospheric pressure room temperature plasma mutagenesis (ARTP), the specific operation steps are as follows:
[0041] The activated wild Bacillus genus XS-4 was picked from the plate and inoculated in the liquid seed medium, cultured at 37 °C and 200 r / min until the middle and late logarithmic growth phase, and 1 mL of the bacterial liquid was taken to measure the OD value. Estimate the bacterial concentration according to the general relationship between OD value and the number of colonies, take 1 mL of the bacterial solution and centrifuge at 8000 r / min for 2 min, collect the precipitate and wash it 2-3 times with normal saline, and dilute it to 10 6 cfu / mL bacterial suspension...
Embodiment 2
[0049] Exploration on the tolerance characteristics of Bacillus licheniformis Mut-148:
[0050]1. Salt tolerance test: Inoculate Bacillus licheniformis Mut-148 into NaCl salt tolerance test medium with salt concentrations of 2%, 4%, 6%, 8%, 10%, 12%, 14%, and 16% respectively , After 24 hours of constant temperature cultivation at 37°C, the OD value was measured at 600nm, and the tolerance curve of the strain to NaCl was drawn. Such as image 3 shown.
[0051] 2. Growth at different temperatures: Bacillus licheniformis Mut-148 was inoculated into NB liquid medium, cultured at 20°C, 30°C, 40°C, and 50°C for 24 hours, and the OD value was measured at 600nm, and the relationship between the strain and the temperature was drawn. Tolerance curves such as Figure 4 shown.
[0052] 3. Growth conditions at different pH: inoculate Bacillus licheniformis Mut-148 into NB liquid medium with pHs of 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0, culture at 37°C for 24 hours, and measure the OD ...
Embodiment 3
[0055] In vitro simulation system to explore the ability of Bacillus licheniformis Mut-148 to decompose protein, starch, cellulose and other macromolecules in a 10% high-salt environment:
[0056] Pick the pure cultured strain of Bacillus licheniformis Mut-148 and inoculate it into beef extract peptone liquid medium with a salt concentration of 100g / L, culture at 37°C for 12 hours, collect the bacteria, prepare a bacterial suspension with an OD value of 0.8, and take 0.1 mL of the bacteria The suspension was inoculated into an Oxford cup inserted in casein medium with a salt concentration of 100g / L, incubated at a constant temperature of 37°C for 48 hours, and the diameter of the hydrolysis circle and the strain diameter were measured. Pick the preferred strain with high protease activity, inoculate it into an Oxford cup inserted in a 100g / L salt concentration starch medium and sodium carboxymethylcellulose medium according to the method described in the casein medium, and cult...
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