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Lactobacillus plantarum (L.plantarum) with capability of inhibiting kidney stone formation and application thereof

A technology of Lactobacillus plantarum and kidney stones, applied to Lactobacillus, bacteria used in food preparation, applications, etc., can solve the problems of questionable safety, unclear mechanism of action, and inquiries, etc., and achieves high safety and ease of use. Colonization, good growth effect

Active Publication Date: 2021-08-27
SICHUAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Recombinant strains can reduce urinary oxalate excretion and oxalate crystal deposition by increasing intestinal oxalate degradation. However, the safety of these genetically modified recombinant bacteria in clinical treatment and probiotic edible products in the future is questionable, and they are also facing relevant regulations. limits
In addition, there are very few other types of natural lactic acid bacteria that have been proved by animal experiments to have the ability to reduce stone formation. Kwak et al. found that after combined treatment of Lactobacillus caseiHY2743 and Lactobacillus caseiHY7201, urine grass Salt excretion decreased, rat kidney crystals decreased, but the physiological characteristics of these two strains have not been reported in detail, and their mechanism of action is also unclear, and their genome sequence information has not been queried in NCBI
In summary, there are few reports on the application of Lactobacillus plantarum isolated from humans and without genetic modification in reducing the formation of renal calcium oxalate stones

Method used

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  • Lactobacillus plantarum (L.plantarum) with capability of inhibiting kidney stone formation and application thereof
  • Lactobacillus plantarum (L.plantarum) with capability of inhibiting kidney stone formation and application thereof
  • Lactobacillus plantarum (L.plantarum) with capability of inhibiting kidney stone formation and application thereof

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Experimental program
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Effect test

Embodiment 1

[0041] Isolation, Screening and Molecular Biological Identification of Lactobacillus plantarum J-15

[0042] 1. Material preparation

[0043] Human feces samples were provided by a healthy person in Chengdu City, Sichuan Province;

[0044] The universal primer pair was synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and the 27F / 1492R sequence is as follows:

[0045] 27F: AGAGTTTGATCMTGGCTCAG

[0046] 1492R: TACGGYTACCTTGTTACGACTT

[0047] MRS liquid medium: glucose 20.0g, peptone 10.0g, beef extract 10.0g, yeast powder 6.0g, Tween 801.0mL, dipotassium hydrogen phosphate 2.0g, ammonium citrate 2.0g, sodium acetate 5.0g, magnesium sulfate 0.58g , manganese sulfate 0.25g, pure water 1L, pH 6.2 ± 0.2 (add 1.6%-1.8% agar for solid medium).

[0048] 2. Specific ways

[0049] Eligible healthy human stool samples. Weigh 1.0 g of the sample, and stir it quickly and fully in a centrifuge tube filled with 9 mL of sterile water to completely disperse the sample. Aspirate...

Embodiment 2

[0052] Detection of main physiological characteristics of Lactobacillus plantarum J-15

[0053] PBS buffer solution: Potassium dihydrogen phosphate 0.20g, disodium hydrogen phosphate 1.15g, sodium chloride 8.0g, potassium chloride 0.2g, add 800mL pure water and stir to dissolve, adjust concentrated hydrochloric acid to the corresponding pH value, and dilute to 1L.

[0054]1. Acid resistance test: the three-generation strain will be activated L. plantarum J-15 was inoculated into PBS buffer solution with pH 4.0, 3.0, and 2.0 with 1% inoculum amount, cultured at 37°C for 3 hours, and the number of viable lactic acid bacteria was determined by gradient dilution plate method.

[0055] 2. Bile salt resistance test: Inoculate the activated third-generation lactic acid bacteria into the MRS liquid medium containing 0.1%, 0.2%, and 0.3% bovine bile salt at an inoculation amount of 1%, respectively, culture at 37°C for 5 hours, and gradually dilute The plate method was used to determ...

Embodiment 3

[0060] In order to ascertain the oxalate degradation characteristics of J-15, the degradation ability of J-15 to sodium oxalate was tested in vitro. The other two strains of lactic acid bacteria y96 and y104 isolated from human feces were tested and compared together.

[0061] Oxalate degradation test: J-15 was inoculated in self-improved TM liquid medium containing 10mmol / L sodium oxalate concentration, cultured at 37°C for 9 days, and detected by ion chromatography (IC) on day 0 and On the 9th day, the oxalate ion concentration in the culture medium calculated the decomposition utilization rate of oxalate, and the ion chromatogram of oxalate concentration was as follows image 3 shown.

[0062] TM liquid medium: 2.0g dipotassium hydrogen phosphate, 2.0g sodium dihydrogen phosphate, 2.0g ammonium sulfate, 0.58g magnesium chloride, 0.25g manganese sulfate, 1L pure water

[0063] Degradation rate (%) = (1-A 1 / A 0 )×100%

[0064] In the formula: A 0 is the initial concent...

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Abstract

The invention relates to the technical field of microorganisms, and particularly discloses lactobacillus plantarum (L.plantarum) J-15 with the capability of inhibiting kidney stone formation and application thereof. The lactobacillus plantarum J-15 provided by the invention is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.22140, and the preservation date is April 6, 2021. The strain is separated from excrement of healthy people in Chengdu of Sichuan province, grows well on an MRS agar culture medium, has certain endurance capacity to acid and cholate, is sensitive to various antibiotics, has certain oxalate degradation capacity and relatively strong renal crystallization inhibition capacity, and is shown by whole genome sequencing to have relatively strong carbohydrate metabolism capacity, membrane transport capacity and environmental adaptability, is applied to the field of functional foods and clinical kidney stone prevention and adjuvant therapy, has actual production value and has very important significance to human health.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of lactobacillus plantarum capable of inhibiting the formation of kidney stones, and in particular to the field of research and development of functional lactic acid bacteria and its products. [0002] technical background [0003] Kidney stones are one of the common urinary system diseases worldwide, which can cause pain, urinary tract infection, chronic kidney disease, and even loss of kidney function, and its prevalence is increasing year by year worldwide. Calcium oxalate stones are the most common type of stones, accounting for about 80%. Only a minority of renal calcium oxalate stones are caused by primary hyperoxaluria, and the cause of renal calcium oxalate stones in most patients remains unclear. Conventional treatment methods include drug therapy, extracorporeal shock wave lithotripsy, and flexible ureteroscopic lithotripsy. Although these treatments can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A23L33/135C12R1/25
CPCC12N1/20C12N1/02A23L33/135A23V2002/00A23V2400/169A23V2200/30
Inventor 孙群田蕾徐小芳崔雅倩张慧
Owner SICHUAN UNIV
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