Lactobacillus gasseri lg08 capable of degrading uric acid and its application
A kind of Lactobacillus gasseri, the technology of action, applied in the field of microorganisms, can solve the problems of whether unreported strains can degrade uric acid, affect the mitigation effect, degrade inosinic acid reports and the like
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Embodiment 1
[0025] Example 1 Screening of strains with degrading uric acid
[0026] 1) strain screening
[0027] Extract 5 mL of diluent from the feces of healthy elderly people in Bama Town, Hechi City, Guangxi Zhuang Autonomous Region, China (the population has not taken antibiotics before collection, no history of taking probiotics, and no history of gastrointestinal disease), and serially diluted 10 times to 10 -6 , 200 μL of each dilution concentration was spread on selective MRS medium, anaerobic culture at 37°C for 48-72h, and different single colonies on the plate were obtained by separation. Using the sterilized inoculation loop, pick the above-mentioned different single colonies on MRS solid medium for streak purification and anaerobic culture at 37°C for 48h to obtain a total of 9 strains of pure lactic acid bacteria, named L1-L9 respectively.
[0028] 2) Analysis of the tolerance of isolated strains to simulated gastric juice
[0029] After collecting and culturing, the cell...
Embodiment 2
[0046] Example 2 Identification of strains
[0047] According to the bacterial strain L8 obtained in Example 1, the identification method is as follows:
[0048] The obtained strain L8 was identified by 16s rDNA sequencing and physiological and biochemical identification. The 16s rDNA universal primer sequence 27F is: 5'-AGAGTTTGATCCTGGCTCAG-3' (Seq ID No: 1), 1492R is: 5'-GGTTACCTTGTTACGACTT-3' (Seq ID No: 2).
[0049] The 16S rDNA gene sequence of strain L8 was amplified and sequenced, and the PCR amplification product was sent to Shanghai Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing. The nucleotide sequence of the 16S rDNA of strain L8 is shown in Seq ID No: 3 . After 16s rDNA gene comparison, the similarity rate with the Lactobacillus gasseri strain in Genebank reached 100%. Combined with the results of physiological and biochemical identification, strain L8 was identified as a Lactobacillus gasseri strain and named Lactobacillus gasseri LG08. Its physiologi...
Embodiment 3
[0054] Example 3 Analysis of Lactobacillus gasseri LG08 degraded nucleosides and nucleotide products
[0055] This embodiment provides a method for detecting the metabolites after Lactobacillus gasseri LG08 degrades adenosine, guanosine, inosine, adenylic acid, guanylic acid, inosinic acid and disodium 5'-guanylate , which includes the following steps:
[0056] Add 5 mmol / L adenosine, guanosine, inosine, adenylic acid, guanylic acid, inosinic acid and disodium 5'-guanylate to the ammonium acetate buffer solution, and the concentration is 0.4g according to the dry weight of the bacteria. After washing the cells with PBS buffer / L, the cells were mixed and cultured at 37°C for 2 hours and then disrupted by ultrasonic wave. The crushing conditions were as follows: put the suspension into an ice-water bath and adopt a strategy of over 3s and 5s (250w), then 4°C, 12000rpm After centrifugation for 15 min, the supernatant was collected and the concentrations of metabolites (hypoxanth...
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