Preparation method and application of culture medium suitable for lactobacillus rhamnosus to produce exopolysaccharide
A technology of Lactobacillus rhamnosus and exopolysaccharides, applied in the direction of microorganism-based methods, biochemical equipment and methods, applications, etc., can solve the problem of difficult removal, low yield of exopolysaccharides of Lactobacillus rhamnosus, and incomplete Eliminate interference and other issues
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Embodiment 1
[0060] Example 1. Acquisition of Lactobacillus rhamnosus strain
[0061] Take 1 mL of fresh milk (fresh milk of New Hope Shuangfeng Milk Cow Breeding Base (Hangzhou)) into a sterile test tube, follow 10 -1 Perform serial dilutions, pipette 10 -2 , 10 -3 , 10 -4 and 10 -5 1ml of the dilution solution was evenly spread on the MRS plate, and placed in a 37°C incubator for 48h. Pick a single colony on the plate, inoculate it on the MRS plate and continue to cultivate for 3 days, inoculate the single colony with signs of acid production into the lactic acid bacteria medium, observe the colony morphology and microscopic morphological characteristics, study the physiological and biochemical changes, and carry out molecular analysis of the bacteria. Biological identification, 40% glycerol was stored at -80 ℃.
[0062] The resulting Lactobacillus rhamnosus ZFM216 activated strain was identified by 16S rDNA sequencing. For strains stored at -80°C, single colonies were selected for...
Embodiment 2
[0065] Embodiment 2, the polysaccharide production situation of Lactobacillus rhamnosus in conventional MRS medium
[0066] 1), the preparation of MRS medium:
[0067] MRS liquid medium: peptone 10g, yeast extract 5g, beef extract 10g, glucose 20g, sodium acetate 5g, diamine hydrogen citrate 2g, Tween-80 1mL, dipotassium hydrogen phosphate 2g, magnesium sulfate heptahydrate 0.2g, Manganese sulfate heptahydrate 0.05g, add ultrapure water to make up to 1L. High temperature sterilization (1.1 atmospheric pressure, sterilization at 115°C for 20min).
[0068] MRS solid medium: 15g / L agar was added to the above-mentioned MRS liquid medium, and the rest were the same.
[0069] 2), the preparation of Lactobacillus rhamnosus MRS fermentation broth:
[0070] Activated strains: Ultraviolet sterilization for 30min, take out the Lactobacillus rhamnosusZFM216 cryopreservation tube stored at -80°C, and thaw at room temperature for use. Take a one-time sterilized inoculation loop and dip ...
Embodiment 3
[0080] Example 3. Preparation of Lactobacillus rhamnosus-producing exopolysaccharide medium
[0081] 1), yeast extract, beef extract and peptone are subjected to polysaccharide removal as follows:
[0082] The yeast extract / beef extract / peptone is fully dissolved in ultrapure water according to the set mass / volume ratio of 10% (yeast extract is 5g / 50mL, beef extract and peptone are 10g / 100mL), and then 4 Double the volume of 4°C pre-cooled absolute ethanol, and place it at 4°C for precipitation for 12 hours; the supernatant was collected by centrifugation at 10,000 rpm for 15 min, and then the ethanol in the supernatant was removed by a rotary evaporator (about 60°C). Corresponding to the yeast extract / beef extract / peptone after depolysaccharide respectively; spare.
[0083] Other conditions are prepared according to MRS liquid medium;
[0084] 2), yeast extract, beef extract and peptone, on the basis of the above polysaccharide removal treatment, the culture time (0-32h), c...
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