Bacillus coagulans and application thereof in mixed fermentation to produce L-lactic acid
A technology of Bacillus coagulans and mixed fermentation, applied in fermentation, bacteria, microorganism-based methods, etc., can solve the problems of low conversion rate and low optical purity of fermentation products, and achieve high sugar-acid conversion rate, shortened fermentation time, and significant Effects of social significance and economic value
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Embodiment 1
[0041] This example illustrates the method of performing low-energy nitrogen ion implantation mutagenesis screening on the original strain of Bacillus coagulans.
[0042] The specific steps for the first step of low-energy nitrogen ion implantation mutagenesis screening are as follows:
[0043] (a) Preparation of single spore suspension: take the original starting strain of Bacillus coagulans BC-3 cultured at 40~45°C for 2~3 days and add 10 mL of sterile water to the fresh slant, scrape and wash the spores and pour them on the glass beads with certain amount Shake well in a 250mL conical flask for 20min (250rpm), break up the spore chains, filter through three layers of degreasing gauze, count the filtrate with a hemocytometer, and adjust the spore concentration by 10 6 individual / ml.
[0044] (b) Low-energy nitrogen ion implantation mutagenesis: take 0.1 mL of the spore suspension in step (a) and evenly spread it on a sterile plate, and perform low-energy nitrogen ion implan...
Embodiment 2
[0059] This example illustrates the genetic stability of high-yield strains Bacillus coagulans BC-N6262 and Lactobacillus casei LC-N235. The results of the subculture fermentation test are shown in Table 2:
[0060] Table 2
[0061]
[0062] From the experimental results, it can be seen that after 5 consecutive passages, the mutant strain Bacillus coagulans BC-N6262 has a relatively stable L-lactic acid production and good passage stability. In addition, the stability of Lactobacillus casei LC-N235 is also ideal. Both can be used as production strains for further research and development.
Embodiment 3
[0064] This example illustrates the production of L-lactic acid by mixed fermentation of Bacillus coagulans BC-N6242 and Lactobacillus casei LC-N235.
[0065] The medium formula described in this embodiment (% is mass percent):
[0066] Plate basic medium: glucose 5.0%, peptone 1.0%, beef extract 0.3%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 3.0%, agar 1.2%, the rest is water, pH 7.0.
[0067] Slant medium: glucose 0.5%, peptone 1.0%, beef extract 0.3%, yeast extract 0.3%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 0.3%, agar 1.2%, the rest is water, pH 7.0.
[0068] Seed medium: glucose 6.0%, peptone 1.0%, yeast extract 1.0%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 3.0%, the rest is water, pH 7.0.
[0069] Fermentation medium: glucose 15%, peptone 0.05%, yeast extract 1.5%, magnesium sulfate heptahydrate 0.05%, heavy calcium carbonate 7.5%, the rest is water, pH 7.0.
[0070] Inoculate the sc...
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