Application of bifidobacterium lactis tci604 and its metabolites
A TCI604, Bifidobacterium lactis technology, applied in the direction of bifidobacteria, applications, metabolic diseases, etc., can solve problems such as complications and patient disease recurrence
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Embodiment 1
[0053] Example 1: Protease activity detection test of Bifidobacterium lactis TCI604
[0054] The cryopreservation tube containing the Bifidobacterium lactis TCI604 strain of [Preparation Example A-3] was thawed, and the TCI604 strain was inoculated into an MRS medium (BDTMDifcoTM Lactobacilli MRS Broth) and cultured at 37°C to activate the strain . Next, take a piece of Skim milk agar and dig a hole in it. Then, 50 microliters of the once activated TCI604 strain was injected into the holes on the aforementioned agar and incubated at 37°C for 24 hours. Finally, observe whether a transparent ring is produced on the cultured agar (if a transparent ring is produced, it means that the cultured strain has protease activity), and take pictures to record. Results are shown in figure 1 .
[0055] Depend on figure 1 It can be seen that Bifidobacterium lactis TCI604 of the present invention can form a transparent ring on skim milk agar. The aforementioned results show that the Bifi...
Embodiment 2
[0056] Example 2: Gastric acid resistance and bile salt resistance test of Bifidobacterium lactis TCI604
[0057] The cryopreservation tube containing the Bifidobacterium lactis TCI604 strain of [Preparation Example A-3] was thawed, and the TCI604 strain was inoculated into an MRS medium (BDTMDifcoTM Lactobacilli MRS Broth) and cultured at 37°C to activate the strain . The activated TCI604 strain was then rinsed twice with 1x phosphate-buffered saline (Phosphate-bufferedsaline, PBS), and then re-dissolved with 0.3 ml of PBS (total bacterial count was about 1x10 10 ). Divide the lysate after the aforementioned treatment into three groups (each group contains 0.1 ml of the lysate), and add each group of lysates to the following solutions:
[0058] 1. Control group: 99 ml of 0.2M KCl solution (pH 7).
[0059] 2. Gastric acid group: 99 ml of 0.2M KCl solution (pH 1.2).
[0060] 3. Bile salt group: 99 ml of 0.2M KCl solution (containing 0.3% bile salt).
[0061] Next, the solu...
Embodiment 3
[0063] Example 3: Intestinal colonization effect test of Bifidobacterium lactis TCI604
[0064] (3-1) The human colon adenocarcinoma cell line Caco-2 was cultured in a triangular dish (T75Flask) for 120 hours. Next, the culture medium was removed, and after rinsing the cells twice with 1x PBS, 1 ml of 1% trypsin (Trypsin / EDTA) was added to suspend the cells. The aforementioned cell suspension was injected into a 24-well plate (4 × 10 5 cells / well) at 37°C, 5% CO 2 Incubate overnight. After that, after rinsing the cells twice with 1×PBS, 900 μl of fresh antibiotic-free culture medium was added to each well.
[0065](3-2) Thaw the cryopreservation tube containing the Bifidobacterium lactis TCI604 strain of [Preparation Example A-3], inoculate the TCI604 strain into MRS medium (BDTMDifcoTMLactobacilli MRS Broth) and culture at 37°C, to activate the strain. The activated TCI604 strain was then rinsed twice with 1x PBS, and then re-solubilized with 1x PBS (total bacterial coun...
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