Preparation method and application of acid-resistant lactobacillus caseiwith high reproductive capacity
A technology of Lactobacillus casei and ability, applied in the field of microorganisms, to achieve enhanced resistance, increased maximum specific growth rate, and improved vitality
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Embodiment 1
[0025] The influence of embodiment 1 exogenous nucleotide 5' disodium guanylate on the growth of Lactobacillus casei
[0026] After the strains preserved in the glycerol protectant were activated in the MRS liquid medium and continuously cultured for 3 generations, the MRS medium was used as a blank control, and 1g / 100mL, 2g / 100mL, 3g / 100mL, 4g / 100mL of 5' Disodium guanylate was inoculated with Lactobacillus casei at a volume ratio of 1%, cultured on a shaker at 37°C for 24 hours, and samples were taken every 4 hours. The live bacteria of Lactobacillus casei were counted by the double dilution method, and the growth curve was drawn. The modified Gompetz model and the modified Logistic model were used to describe the growth and variation of Lactobacillus casei nonlinearly.
[0027] (1) Modified Gompertz model
[0028]
[0029] In the formula, t is time (h), N(t) is the number of bacteria at time t, N max , N 0 is the maximum and initial bacterial count (CFU / g). mu max i...
Embodiment 2
[0035] The influence of embodiment 2 exogenous nucleotides on Lactobacillus casei biofilm, extracellular polymer
[0036] Lactobacillus casei (1% inoculum) was inoculated in MRS broth culture medium for 24 hours on a shaker (37°C, 160r / min), diluted at a ratio of 1:100, and 100 μL of bacterial solution and 4.0% 5'guanosine Disodium acid MRS medium was added to the well plate, and MRS broth medium was used as a control. Put sterile coverslips and polished zinc sheets (0.5mm×0.5mm×0.3mm) into the orifice plate, and after standing at 37°C for 48 hours, wash the coverslips slowly with sterile water for 3 times, crystal violet ( 0.4%) was stained for 20 minutes, and the changes of the bacterial biofilm were observed under a microscope. Rinse the zinc sheet slowly with sterile water to remove non-adherent bacteria. Fix with 2.5% glutaraldehyde for 30 min and then dry for 30 min. Raman spectroscopy was used to detect the effect of exogenous nucleotides on the extracellular polymer...
Embodiment 3
[0039] Example 3 Molecular docking study on the mechanism of action of disodium 5' guanylate
[0040] The biofilm and extracellular polymers of Lactobacillus casei can be increased by inducer-2 (AI-2). AI-2 is a signal molecule synthesized through the methyl cycle. Lux S protein is the key enzyme for the synthesis of AI-2 , so it is of great significance to study the interaction between disodium 5'guanylate and lux S. Molecular docking is a theoretical simulation method that mainly studies the interaction between molecules (such as ligands and receptors) and predicts their binding mode and affinity. In recent years, molecular docking has become an important technology in the field of computer-aided drug research. In this example, the interaction between 5' disodium guanylate, AI-2 type signal molecule and the active site of Lactobacillus casei LuxS receptor protein was studied. The result is as Figure 4 As shown, Figure A is the interaction between AI-2 signal molecules and...
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