Lactobacillus zjuids03 against Helicobacter pylori and its application
A Helicobacter pylori, antagonistic technology, applied in the field of microorganisms, can solve the problems of poor inhibition/inactivation effect of Helicobacter pylori, etc.
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Embodiment 1
[0055] Embodiment 1, screening and identification of Lactobacillus paracasei ZJUIDS03:
[0056] 1. Screening of Lactobacillus paracasei ZJUIDS03
[0057] 1.1 Sample source
[0058] The bacterial strains used in the present invention are all collected from 10 copies of traditional fermented products in Yunnan --- farmhouse-made milk fans; that is, the present invention uses farmhouse-made milk fans from Yunnan as samples.
[0059] 1.2 Preparation of medium
[0060] The medium used for sample isolation and strain screening was MRS solid medium, which was purchased from Beijing Luqiao Company. The composition of MRS solid medium is shown in Table 1; the "20g agar" in Table 1 was canceled to obtain MRS liquid medium.
[0061] Use deionized sterile water as a solvent, adjust the pH of the dissolved medium to 6.2-6.4, sterilize with a pressure cooker at 121°C for 15 minutes, pour the unsolidified MRS solid medium into a sterile culture plate at about 55°C, Let stand until the med...
Embodiment 2
[0077] Embodiment 2, co-cultivation of Lactobacillus paracasei ZJUIDS03 inhibits Helicobacter pylori ATCCA43504
[0078] 1. Culture of Helicobacter pylori
[0079] The glycerol tube containing Helicobacter pylori ATCCA43504 was taken out from -80°C, and spread on Columbia blood agar medium containing 5% (v / v) sheep blood for activation culture. The culture conditions were microaerobic environment, 37°C , cultivated for 72h-96h.
[0080] After being activated for one generation in a solid medium (Columbia blood agar medium containing 5% sheep blood), pick a single colony and continue to streak for two generations. Pick a single colony after purification and culture and insert it into the Helicobacter pylori liquid medium , the liquid culture condition is microaerophilic environment, 37°C, and the time is 48h-72h. 8000rmp / min, centrifuge at 4°C for 15min to collect supernatant and precipitate respectively. Wash the pellet twice with clean Helicobacter pylori liquid medium, an...
Embodiment 3
[0100] Embodiment 3, Lactobacillus paracasei ZJUIDS03 inhibit the growth of Helicobacter pylori ATCCA43504
[0101] 1. Culture of Helicobacter pylori
[0102] With the step 1 of embodiment 2.
[0103] 2. Culture of lactic acid bacteria
[0104] Wash the lactic acid bacteria pellet twice with high-temperature sterilized PBS, and resuspend to make the concentration of viable bacteria reach 10 11 CFU / mL; the rest are equal to step 2 of Example 2.
[0105] 3. Growth Inhibition Experiment
[0106] 100 μl of Helicobacter pylori bacteria solution (10 9 CFU / mL) was applied to a flat plate (about 20ml) of Columbia blood agar medium containing 5% sheep blood without antibiotics and punched, and any of the following was added respectively to form different experimental groups:
[0107] 100 μl of lactic acid bacteria suspension to be tested, 100 μl of lactic acid bacteria supernatant to be tested; 100 μl of positive control (0.025% metronidazole solution); negative control (100 μl of...
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