Fermentation culture method for bacillus subtilis PTS-394
A technology of PTS-394 and Bacillus subtilis, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problem of low bacterial content and achieve the effect of increasing bacterial content
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Embodiment 1
[0013] Example 1 Determine the carbon source and nitrogen source of the culture medium
[0014] The basic fermentation medium of Bacillus subtilis PTS-394 (disclosed in Example 3 in the patent application with publication number CN102550607A): 20 g corn flour, 10 g soybean meal, 5 g soybean meal, 1.5 g yeast powder, CaCO 3 3 g, NaCl 0.5 g, fish meal 10 g, make up to 1000 mL, pH 7.5.
[0015] Replace the 20 g / L corn flour in the above basic fermentation medium with sucrose, soluble starch, corn starch, maltose, lactose, and xylose in the same amount (20 g / L) of 6 different carbon sources to form different carbon sources. base. Replace the 10 g / L soybean meal powder of the basic fermentation medium with soybean meal, tryptone, yeast extract powder, beef extract, peptone, and fish meal in equal amounts (10 g / L) of 6 different nitrogen sources, and mix them into different nitrogen sources. Medium.
[0016] Adjust the pH of the culture media with different carbon and nitrogen sources t...
Embodiment 2
[0021] Example 2 Determine the key factors of fermentation medium composition
[0022] Using Plackett-Burman experimental design, from Example 1 Bacillus subtilis PTS-394 medium components (corn starch, soybean powder, yeast powder, CaCO 3 , NaCl, fish meal) key factors were screened out of 6 factors. The PB experiment design is shown in Table 2:
[0023] Table 2 Plackett-Burman experimental design and results of medium component factors
[0024]
[0025] Note: In the experiment, each factor takes two levels. The high level "1" means that the content of the component is 1.5 times that of the initial fermentation medium; the low level "-1" means that the content of the component is 0.5 times that of the initial fermentation medium.
[0026] Regression analysis is performed on the data in Table 2, and the main factor coding equation is:
[0027] Y 0 =9.47+0.12A+0.055B-0.032C+0.020D+9.095E-003E-0.012F
[0028] Where: Y 0 Is the predicted value of bacteria content (cfu / mL) (the same below...
Embodiment 3
[0032] Example 3 Optimal ratio of key factors of fermentation medium
[0033] Analyzing the results of the PB experiment, we can see that the three factors of corn starch, soybean meal and yeast powder are the key factors affecting the content of Bacillus subtilis PTS-394, according to the fitting equation Y 0 Each variable coefficient determines the climbing direction and step length of the key factors, and the steepest climbing experiment is carried out. The climbing experiment design and results are shown in Table 4:
[0034] Table 4 The steepest climbing experiment design and results of the key factors of the medium composition
[0035]
[0036] It can be seen from Table 4 that among the combination of 3 factors, the number 4 combination has the largest bacterial content, so this combination is selected as the central value of the response surface experiment, and the next RSM experiment design is carried out.
[0037] According to the results of the steepest climbing experiment (T...
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