Bifidobacterium bifidum bxm0 and its application
A Bifidobacterium bifidum and microbial agent technology, applied in Bifidobacterium bifidum BXM0 and its application fields, can solve problems such as large side effects and inability to be widely used
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Embodiment 1
[0049] Embodiment 1: the preparation of the identification of Bifidobacterium bifidum BXM0
[0050] 1.1 Identification of Bifidobacterium bifidum BXM0 strain
[0051] The feces samples of healthy adults living in the urban area of Beijing for a long time were collected, diluted and spread on YCFA medium, cultured anaerobically at 37°C for 24-48 hours, and isolated different single colonies on the plate. Use a sterilized inoculation loop to pick different single colonies and streak them on a new YCFA solid medium plate for purification, and culture them anaerobically at 37°C for 48 hours to obtain purified single colonies. Each purified single colony was spread on a mass spectrometer plate, and the lysate and matrix were added respectively. After drying, mass spectrometry was carried out on a MALDI-TOF MS 1000 mass spectrometer (Autobio, Zhengzhou Antu Biotechnology Co., Ltd.). After the identification was completed, each strain was frozen at -80°C in the strain resource ban...
Embodiment 2
[0054] Embodiment 2: paper chromatography detects GABA
[0055] Take 1 mL of Bifidobacterium bifidum BXM0 fermentation broth and centrifuge at 12,000 rpm for 10 min at 4°C to remove the bacteria, and transfer the supernatant to a new centrifuge tube for future use. Pipette 4 μL of sample supernatant (set for 2 repetitions), corresponding culture medium and any concentration of GABA standard solution (0.2, 0.4, 0.8, 1.2, 1.6 and 2.0 g / L) to spot on the filter paper, the spot line distance The edge of the chromatography paper is 2.0 cm, and the spacing between samples is 2.0 cm. After the sample spot is dried, put the chromatographic paper in the developer (V (n-butanol): V (glacial acetic acid): V (water) = 5: 3: 2, the amount of ninhydrin is 1.2% (w / v)) Develop in medium for 50 min (when the developer is about 1 cm away from the upper edge of the chromatography paper). Immediately after the chromatography, the chromatography paper was dried in an oven at 90°C for 30 min to d...
Embodiment 3
[0059] Embodiment 3: High performance liquid chromatography HPLC method detects bacterial liquid GABA
[0060] Accurately weigh GABA, prepare GABA standard solutions with mass concentrations of 0.04, 0.08, 0.12, 0.16 and 0.20 g / L, take 100 μL of GABA standard solutions of each concentration, and add 100 μL of OPA (phthalaldehyde) derivative Mix well and incubate at room temperature for at least 90s. After the reaction, 20 μL of each solution was injected, and the peak absorption was measured at 338 nm. The standard curve of GABA concentration detection was established by the peak area. Then, add OPA (phthalaldehyde) derivative to the diluted BXM0 supernatant, incubate and mix well, inject the sample, and measure the absorption peak at 338 nm.
[0061] Bifidobacterium bifidum BXM0 was cultured in YCFA liquid medium (without additional L-glutamic acid) for 24 hours, and the amount of GABA that could be produced by HPLC was 0.273g / L. It is worth mentioning that various concent...
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