Preparation method for pediococcus acidilactici strain freeze-drying preparation
A drying technology of Pediococcus lactis, which is applied in the direction of preservation of microorganisms, etc., can solve problems such as difficulties in large-scale industrial production of Pediococcus lactis strains
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Embodiment 1
[0033] Preparation of freeze-dried preparation of Pediococcus lactis strain
[0034] (1) Inoculate the Pediococcus lactis PA003 strain stored on the MRS slant at 4°C in the liquid MRS medium at an inoculation amount of 1%, and culture it at 37°C for 14 hours after inoculation, and then press the liquid MRS suspension 1% of the inoculum was inoculated in liquid growth medium and incubated at 37°C for 14 hours.
[0035] (2) Centrifuge with a refrigerated centrifuge (10000rpm, 4°C, 10min), discard the supernatant, and collect the solid precipitate (cells); wash twice with the same volume of sterilized Ringer's solution, discard the supernatant, and collect the solid precipitate (cells); after that, suspend the cells in the sterilized freeze-drying protective agent medium, let it stand at 25°C for 1 hour, then let it stand at 4°C for another 1 hour, and finally place it at -20°C , ready for freeze-drying after freezing.
[0036] (3) The frozen sample was placed in a freeze dryer...
Embodiment 2
[0038] Survival rate of freeze-dried preparation of Pediococcus lactis strain
[0039](1) Viable bacterial counts of Pediococcus lactis strains before freeze-drying: After standing at 25°C for 1 hour above, take 1 mL of the bacterial suspension of the protective agent from the freeze-drying protective agent medium and add it to 9 mL of sterilized physiological In brine, after proper dilution, it reaches 10 -6 、10 -7 At this time, take 100 μL of the bacterial suspension under the dilution gradient and add it to the MRS solid medium plate for colony coating (each two adjacent gradients are coated with 3 plates), and the coated plates are placed at 37 ° C. After culturing for 48 hours, plate colonies were counted. The number of surviving colonies is expressed in cfu / mL.
[0040] (2) Immediately after freeze-drying, refold the freeze-dried preparation with the same volume of refolding solution to form a refolded bacterial suspension under refolding conditions. Take 1 mL of bac...
Embodiment 3
[0043] Antibacterial activity of freeze-dried preparation of Pediococcus lactis strain
[0044] Inoculate the freeze-dried Pediococcus lactis PA003 from the refolding solution into the MRS solid culture dish, and culture it at 37°C for 24 hours, until it grows colonies visible to the naked eye; insert it into 5mL semi-solid TSYEB at about 50°C 1 μL of Listeria mononucleosis CVCC1595 was mixed and poured into the above-mentioned MRS solid culture dish that had grown visible colonies to the naked eye, so that the layer of semi-solid medium was spread on the solid MRS medium, and then Place the plate at 37° C. and incubate for 12 hours, and observe whether a bacteriostatic zone appears around the Pediococcus lactis colony. ( Figure 12 )
[0045] Depend on Figure 12 It can be seen that after the lyophilized preparation made was refolded, the bacteriostatic test was carried out to the surviving Pediococcus lactis PA003, and it was found that a clear bacteriostatic zone could b...
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