Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof
A technology of antibacterial activity and lactic acid bacteria, applied in the field of microorganisms, can solve problems such as intolerance to bile salts and failure to grow, and achieve good antibacterial effect
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Embodiment 1
[0035] Example 1 Screening, domestication and isolation of Lactobacillus salivarius (L.salivarius) GZPH2
[0036] Randomly buy kimchi samples from the Guangzhou market, take 1mL of sample liquid and add 50mL of aseptically treated MRS liquid (adding 0.05% CaCO 3 ), 150r / min 37 ℃ enrichment culture for 12h; the enrichment solution was diluted to 10 with sterile physiological saline -8 , spread each dilution gradient on the plate in turn, culture at 37°C for 24 hours, select a plate with a single colony number of 30-300, and select a single colony with obvious calcium carbonate dissolution circle around the colony, and pick the corresponding single colony to the MRS solid medium with the inoculation needle Three districts are cross-marked until the single bacterial colony that occurs is uniform in size, and Gram staining is selected as purple (positive), and the single bacterial colony with uniform shape and size of the stained bacterium is initially confirmed as lactic acid bac...
Embodiment 2
[0049] The broad-spectrum antibacterial activity of embodiment 2 Lactobacillus salivarius (L.salivarius) GZPH2
[0050] (1) Preparation of 39 strains of indicator bacteria
[0051] Inoculate a single colony of 39 strains of indicator bacteria (Table 1) into the nutrient broth liquid medium, culture at 37°C, 150r / min for 8h, and adjust its concentration to 1×10 6 CFU / mL, stored at 4°C, for later use;
[0052] Table 1 39 different experimental indicator strains
[0053]
[0054]
[0055] (2) Preparation of lactic acid bacteria
[0056] Inoculate the single colony of Lactobacillus salivarius (L.salivarius) GZPH2 identified in Example 1 in 50mL MRS liquid medium, 37°C, 150r / min, cultivate for 12h, and cultivate its concentration to 1×10 8 CFU / mL; 10000RCF, 4°C, centrifuge the lactic acid bacteria culture solution for 10 minutes, take the supernatant for the antibacterial test, store at 4°C, and set aside;
[0057] (3) (Supernatant) antibacterial activity screening
[00...
Embodiment 3
[0067] Other characteristic analysis of embodiment 3 Lactobacillus salivarius (L.salivarius) GZPH2
[0068] (1) Hemolysis analysis
[0069] Get 5 μ L seed liquid (final concentration is 8.0 * 10 8 CFU / mL) was spotted on a blood plate (sheep blood, Guangdong Huankai Biological Co., Ltd.), cultured at 37°C for 24 hours, and observed whether there was hemolysis around the colony. Select No. 8 vibrio parahaemolyticus (Vibrio parahaemolyticus) as positive control;
[0070] (2) Histamine detection
[0071] Prepare nutrient broth liquid medium, containing mixed amino acids (histidine 10.0g / L), adjust the pH of the medium to 5.5, add bromocresol purple (0.06g / L), agar powder (20g / L, gel strength >1300g / cm 2 ), sterilized at 120°C for 15min and poured onto the plate; activate the identified Lactobacillus salivarius (L.salivarius) GZPH2 bacterial strain in Example 1, after appropriate dilution, draw 100 μ L to coat the plate, cultivate it at 37°C for 48h, and make a positive control...
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