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A highly efficient purine-degrading Lactobacillus rhamnosus yzulr026 and its application

A technology of Lactobacillus rhamnosus and purine, applied to Lactobacillus rhamnosus strains and the application field of degrading purines, can solve the problem of rare strains of purine substances, achieve rapid and efficient degrading ability of guanine, reduce guanine Absorption, good acid resistance

Active Publication Date: 2022-07-08
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many strains of Lactobacillus rhamnosus have been used to prepare microecological preparations such as food starters and intestinal balance regulators. However, strains suitable for rapid degradation of purine substances in vivo and in vitro are still very rare

Method used

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  • A highly efficient purine-degrading Lactobacillus rhamnosus yzulr026 and its application
  • A highly efficient purine-degrading Lactobacillus rhamnosus yzulr026 and its application
  • A highly efficient purine-degrading Lactobacillus rhamnosus yzulr026 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Screening of efficient purine-degrading strains

[0036] 50 strains were isolated from traditional Tibetan fermented foods (yogurt, pickles, sweet fermented grains), after 3 times of activation, inoculated with MRS medium, anaerobic culture at 37 °C for 24 h, taking 1 mL of 1 × 10 7 CFU / mL bacterial culture solution was centrifuged at 8000 r / min at 4°C for 10 min, and the bacterial cells were collected. Resuspend and wash the cells 2-3 times with 1 mL of sterile ultrapure water. Add 750 μL of 20 μg / mL reaction solution of xanthine, hypoxanthine and guanine to the bacterial cells, add ultrapure water to the control sample, mix by vortex, incubate at 37°C for 2 hours at 120 r / min, and determine by liquid chromatography Purine content in the sample.

[0037] The specific measurement process is as follows: sampling after culture, centrifuging at 8000 r / min for 10 min at 4°C to take the supernatant; filtering the supernatant through a 0.22 μm microporous membrane, and meas...

Embodiment 2

[0040] Guanine degradation characteristics of strain YZULr026

[0041] 100 μL of strain YZULr026 preservation solution (log phase) was inoculated into 5 mL of MRS liquid medium, cultured anaerobic at 37 °C for 24 h, and successively passaged for 3 times. The number of viable bacteria in the culture solution (CFU / mL) was determined by plate dilution coating method. The activated strain culture solution was inoculated into fresh MRS liquid medium at a volume ratio of 2%, and cultured anaerobic to a stationary phase. Adjust the concentration of YZULr026 bacterial solution to 10 7 Take 1 mL of CFU / mL into a centrifuge tube, centrifuge at 8000 r / min at 4°C for 10 min, collect the bacteria, and wash with ultrapure water 2-3 times. (1) Add 750 μL of 20 μg / mL purine reaction solution to the bacterial cells, mix by vortexing, incubate at 37°C with shaking at 120 r / min for 5 hours, and sample and analyze at 0, 1, 2, 3, 4, and 5 hours respectively; (2) Add purine reaction solution with...

Embodiment 3

[0044] Physiological and biochemical identification of strain YZULr026

[0045] The obtained strain YZULr026 was streaked and inoculated into MRS solid medium, anaerobic culture at 37°C for 24 hours, single colony smears on the plate were picked, fixed, dripped with crystal violet staining solution, stained for 1 min, washed with water; dripped with iodine solution, Act for 1 min, wash with water; add 95% ethanol dropwise for 15-20 s, wash with water; counterstain with safranin for 1 min, wash with water; microscopically examine and record the cell morphology of the strain. At the same time, single colony was picked for catalase test, indole, hydrogen sulfide and ammonia, nitrate reduction, arginine hydrolysis, and sugar fermentation test.

[0046] The results showed that the strain was Gram-positive bacilli ( image 3 ), catalase negative, does not produce indole, hydrogen sulfide and ammonia, does not reduce nitrate, does not hydrolyze arginine, can ferment glucose, rhamnos...

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Abstract

The present invention discloses a strain of Lactobacillus rhamnosus YZULr026 that efficiently degrades purines and an application thereof. The strain is deposited in CCTCC, China Type Culture Collection Center, preservation number: CCTCC NO: M 2020359, preservation date: July 27, 2020 . The Lactobacillus rhamnosus YZULr026 of the invention has good acid resistance and bile salt resistance; it has the ability to efficiently degrade purine, and the guanine degradation rate is as high as 87.85% when the strain and guanine are incubated in a 37° C. pure water system for 2 hours; YZULr026 has a unique Genome structure, in which the highest homology between the gene sequence Gene264 encoding α-galactosidase and the published sequence is 93.38%, and there is a 28-base difference; the strain YZULr026 is used to prepare microbial preparations for in vitro and in vivo reduction in food Purine content and prevention of hyperuricemia have broad application prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a strain of Lactobacillus rhamnosus with the ability to efficiently degrade purines and its application in degrading purines. Background technique [0002] Purine is an important base that constitutes nucleic acid. It participates in human metabolism in the form of purine nucleotides, and has important functions such as energy supply, metabolic regulation, and composition of coenzymes. Due to the lack of uric acid oxidase in the human body, purine-containing substances (guanylic acid, guanosine, guanine, etc.) in the body are eventually catabolized into uric acid. Hyperuricemia (Hyperuricemia, HUA). People with hyperuricemia should strictly control their diet and reduce exogenous purine intake. Studies have shown that purine-rich foods include meat, poultry, fish, seafood, eggs, soybeans and other high-protein foods. Strictly controlling the intake of such foods will not only cause n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20A61K35/747A61P19/06C12R1/225
CPCC12N1/20A23L5/28A61K35/747A61P19/06A23V2400/175
Inventor 杨振泉刘慧敏郑香峰高璐饶胜其
Owner YANGZHOU UNIV
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