Lactobacillus plantarum JM113 and application thereof
A technology of Lactobacillus plantarum and JM113, applied to Lactobacillus plantarum JM113 and its application field, can solve the problems of inconsistency and complex probiotic results, small bacteriostatic effect, etc., and achieve the effects of improving production performance and good fermentability
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Embodiment 1
[0026] Embodiment 1: the isolation of plantarum lactobacillus JM113
[0027] 1. Strain samples: Tibetan chickens in Tibet are currently mainly raised in traditional free-range methods. They have strong disease resistance and have not been exposed to antibiotics during their growth. They can be used as an ideal source of probiotics. The samples were collected from the intestines of free-range broilers in Yangling, Shaanxi, and the contents of the intestines and mucous membranes were stored at 4°C.
[0028] 2. Culture medium: MRS medium (ManRogosaSharp, MRS), lactobacillus selective medium, the composition is as follows: trypsin 10g / L, yeast extract 5g / L, beef extract 10g / L, sodium acetate trihydrate 5g / L , glucose 20g / L, Tween 801ml / L, dipotassium hydrogen phosphate 2.0g / L, triamine citrate 2.0g / L, magnesium sulfate heptahydrate 0.58g / L, manganese sulfate monohydrate 0.25g / L, calcium carbonate 20g / L, agar 15g / L, distilled water 1L.
[0029] 3. Isolation and purification of ba...
Embodiment 2
[0037] Embodiment 2: the research of acid resistance and bile salt resistance of Lactobacillus plantarum JM113
[0038] 1. First prepare MRS medium with a pH of 2.5 and MRS medium with a bile salt concentration of 0.2%, and sterilize at 121°C for 20 minutes; then pick a single colony of Lactobacillus and add it to 2ml of MRS medium with a bile salt concentration of 0.2% , cultivate overnight at 37°C; then vortex the lactic acid bacteria solution for 30s, take 150μl of the same bacteria solution and transfer it to 4850μl sterilized normal saline, and make a 10-fold gradient dilution with sterilized normal saline (take out 500μl from the first tube and add to 4500μl In sterilized saline, that is, diluted 10 times, each tube must be vortexed for 30 seconds before taking the liquid), and gradually diluted to 10 -6 , take 500 μl to smear on the plate, do 3 repetitions for each bacterium, place the plate in a 37°C incubator for 24 hours, count the number of colonies M after the cult...
Embodiment 3
[0040] Embodiment 3: the research of resistance to trypsin of Lactobacillus plantarum JM113
[0041] (1) Prepare MRS liquid medium with a trypsin concentration of 1 mg / ml (MRS medium is sterilized at high temperature, add trypsin after cooling, dissolve trypsin in sterile water, and filter with a filter membrane)
[0042] (2) Take 150 microliters of the lactic acid bacteria liquid cultured overnight, and after 10-fold gradient dilution with normal saline, smear the plate and count, and record the number of colonies as M; In MRS liquid culture medium, culture at 37°C for 4 hours, and after 10-fold gradient dilution with normal saline, plate the plate and count, and record the number of colonies as N.
[0043] (3) Count the growth rate n=M / N of Lactobacillus plantarum 113
[0044] (4) Experimental results: the initial number of colonies M is 1.83×10 8 CFU
[0045] The number of colonies N=1.61×10 after 4 hours of trypsin treatment 8 CFU
[0046] Survival rate n=88%
[0047...
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