A kind of liquid mixed fermentation method of clostridium butyricum and bacillus coagulans
A technology of Bacillus coagulans and Clostridium coagulans, which is applied in the field of liquid mixed fermentation of Clostridium butyricum and Bacillus coagulans, can solve the problem of not finding mixed fermentation of liquid multi-bacteria, and achieve the effect of reducing production costs
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Embodiment 1
[0042] Embodiment 1 strain growth curve is measured
[0043] Clostridium butyricum is cultivated in the deep static method of liquid, the coefficient of liquid in the conical flask is 60%, the inoculum size is 4%, sealed with 8 layers of gauze and 2 layers of kraft paper, and cultured at 37°C.
[0044] Bacillus coagulans culture method: Erlenmeyer flask filling fluid coefficient is 20%, inoculum size is 4%, cultivated at 37° C. and 180 rpm.
[0045] According to the above method, the bacteria were activated and cultured. Clostridium butyricum was sampled every 4 hours between 0h and 48h to measure the OD600 value, and Bacillus coagulans was sampled every 2h between 0h and 24h to measure the OD600 value, and their growth curves were drawn respectively. Simultaneously measure the pH value of the bacterial solution. The growth and pH curves of Clostridium butyricum and Bacillus coagulans are as follows: figure 1 with figure 2 .
[0046] Depend on figure 1 It can be seen tha...
Embodiment 2
[0048] Example 2 Mixed Fermentation Medium Composition Optimization
[0049] Mixed culture initial fermentation method: Erlenmeyer flask filling fluid coefficient is 40%, initial inoculum amount of Clostridium butyricum is 6%, initial inoculum amount of Bacillus coagulans is 4%, static culture at 37°C, Erlenmeyer flask with 8 layers of gauze and 2 layers of kraft paper sealing.
[0050] Glucose, sucrose, maltose, lactose, bran, soluble starch, corn starch, etc. were selected as alternative carbon sources with a concentration of 10 g / L. Tryptone 10g / L and yeast extract 5g / L were used as the initial nitrogen source.
[0051] Using the selected optimized carbon source, select the optimal nitrogen source from corn steep liquor powder, soybean meal powder, fish meal, beef extract, peptone, tryptone, yeast extract, and yeast extract powder, and the addition amount is 15g / L.
[0052] Orthogonal experiment was designed according to the optimization results of carbon source and nitro...
Embodiment 3
[0088] Embodiment 3. Optimization of mixed fermentation culture conditions
[0089] On the basis of the optimal fermentation medium, the culture conditions of the mixed fermentation were optimized.
[0090] Fill a 250mL Erlenmeyer flask with 40% volume of medium, the initial pH is 7.0; after sterilization and cooling, insert 8% Clostridium butyricum and 4% Bacillus coagulans seeds, and test the bacteria after standing at 37°C for 36 hours concentration.
[0091] (1) The fermentation temperature is 28°C, 33°C, 37°C, 40°C, 45°C, and the influence of the fermentation temperature on the fermentation is investigated. The results are as follows: Image 6 shown. Depend on Image 6 It can be seen that the temperature has a great influence on the growth of the strains during the mixed fermentation process. When the temperature is in the range of 28-37°C, as the temperature rises, the concentrations of Clostridium butyricum and Bacillus coagulans increase gradually, and at 37°C The ...
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