Lactobacillus rhamnosus culture medium and culture method
A technology of Lactobacillus rhamnosus and culture method, which is applied in the field of culture medium and culture of Lactobacillus rhamnosus, can solve the problems of few studies on the proliferation and culture of strains, achieve cost reduction, increase the number of viable bacteria, and facilitate enrichment Effect
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[0042] 1. Preparation of strains for fermentation
[0043] 1) streak culture: after the Lactobacillus rhamnosus bacteria powder is diluted with sterile water, streak four areas on the MC medium, after the streak is completed, the plate is inverted, and cultured at a constant temperature of 37 ° C in an incubator for 48 to 72 hours;
[0044] 2) Primary purification culture: pick a single colony with a large calcium-dissolving circle on the plate and inoculate it into a test tube containing 5 ml of MRS liquid medium, seal the test tube, and culture at a constant temperature in a 37°C incubator for 17 to 24 hours;
[0045] 3) Secondary purification culture: according to the inoculum amount of 3 to 5%, inoculate the cultured primary bacterial suspension into the triangular flask containing the MRS liquid medium, and the volume of the medium filling accounts for 40% of the volume of the triangular flask, The triangular flask is sealed, and the incubator is kept at a constant temperat...
Embodiment 1
[0056] Embodiment 1 Lactobacillus rhamnosus culture medium and culture method
[0057] 1. Lactobacillus rhamnosus medium, including basal medium and optimized medium.
[0058] Formulation and preparation of basal medium: yeast peptone 8.0g / L, glucose 25.0g / L, yeast extract 3.0g / L, sodium acetate 6.0g / L, magnesium sulfate 0.5g / L, Tween 80 0.5g / L , Isomalt oligosaccharide 1.0g / L, potassium dihydrogen phosphate 3.0g / L, the balance is sterile water, pH value 6.20; weigh each component according to the formula ratio, mix, heat and dissolve, use 1mol / L food grade The pH value of the medium was adjusted to 6.20 with NaOH solution, and sterilized at 115 °C for 30 min.
[0059] The formulation and preparation of the optimized medium: yeast peptone 14.0g / L, glucose 30.0g / L, yeast extract 6.0g / L, sodium acetate 3.0g / L, magnesium sulfate 0.10g / L, Tween 80 0.5g / L , isomalt oligosaccharide 2.0g / L, potassium dihydrogen phosphate 3.0g / L, the balance is sterile water, pH value 6.80; weigh ea...
Embodiment 2
[0076] Embodiment 2 is used for the selection experiment of the strain preservation mode of fermentation
[0077] After Lactobacillus rhamnosus is cultured by streaking, primary and secondary purification, the bacterial suspension is frozen and stored in the slant, bacterial suspension and the preservation mode of the centrifuged bacteria mud in Example 1, and then the three The strains frozen in different preservation methods were fermented under the same culture conditions, and the viable count and yield of the bacterial suspension were determined respectively after the fermentation.
[0078] Determination method of the number of viable bacteria: plate counting method;
[0079] Yield determination method:
[0080]
[0081] Table 1 The effect of fermentation on yield and viable count of bacteria using different preservation methods
[0082] storage method
[0083] From the data in Table 1, it is shown that the bacterial species preserved by the centrifugal bact...
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