Oxalic acid degrading bacterium NJODL1 and application thereof

A technology for degrading bacteria and oxalic acid, applied in the biological field to achieve the effects of reducing the concentration of oxalic acid in urine, excellent acid resistance and bile salt resistance, and good colonization ability

Active Publication Date: 2013-09-11
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, isolating probiotic lactic acid bacteria capable of degrading oxalic acid from dogs with a low incidence of calcium oxalate stones can be used to degrade oxalic acid and regulate oxalate metabolism in animals, which is a good biological control method for calcium oxalate stones in dogs

Method used

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  • Oxalic acid degrading bacterium NJODL1 and application thereof
  • Oxalic acid degrading bacterium NJODL1 and application thereof
  • Oxalic acid degrading bacterium NJODL1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Isolation and identification of Lactococcus lactis lactic acid subspecies

[0041] 1. Preparation of culture medium

[0042] The formula of MRS liquid medium is: 1L distilled water contains: peptone 10g, yeast extract 5g, glucose 20g, beef extract 10g, sodium acetate 5g, triammonium citrate 2g, Tween 801mL, MgSO 4 ·7H 2 O0.58g, MnSO 4 4H 2 O0.05g, K 2 HPO 4 2g, adjusted to pH = 6.2-6.4; sterilized by high-pressure steam at 121°C for 15 minutes, and set aside.

[0043] 2. Sample source

[0044] Three breeds of dogs that are not prone to canine urolithiasis were selected in a certain dog farm. There were eight dogs of each breed, half male and half female. All dogs were fed with commercial dog food (FOSS dog food, Nanjing Police Dog Research Institute Science and Technology Development Center Animal Food Factory). None of the selected dogs had a history of changing dog food, had not used antibiotics within three months, and had not experienced diarrhea....

Embodiment 2

[0059] Embodiment 2: the expanded culture of the isolated bacterial strain NJODL1 of embodiment 1

[0060] The formula of the seed medium is, 1L of distilled water contains: peptone 10g, yeast extract 5g, glucose 20g, beef extract 10g, sodium acetate 5g, triammonium citrate 2g, Tween 801mL, MgSO 4 ·7H 2 O0.58g, MnSO 4 4H 2 O0.05g, K 2 HPO 4 2g, adjusted to pH = 6.2-6.4; sterilized by high-pressure steam at 121°C for 15 minutes, and set aside.

[0061] The bacterial strain NJODL1 obtained in Example 1 was inoculated in a 250mL Erlenmeyer flask equipped with 50mL seed medium, and cultivated for 24h at a temperature of 37°C to obtain a liquid culture of bacterial strain NJODL1, and the number of cells was 3.2×10 9 CFU / mL.

Embodiment 3

[0062] Embodiment 3: Utilize bacterial strain NJODL1 to ferment and degrade oxalic acid in the MRS fermentation medium containing 20mmol / L oxalic acid

[0063] The formula of oxalic acid MRS fermentation medium is: 1L distilled water contains: peptone 10g, yeast extract 5g, glucose 20g, beef extract 10g, sodium acetate 5g, triammonium citrate 2g, Tween 801mL, MgSO 4 ·7H 2 O0.58g, MnSO 4 4H 2 O0.05g, K 2 HPO 4 2g, 2.68g sodium oxalate, adjust pH=6.2-6.4; sterilize by high-pressure steam at 121°C for 15min.

[0064] The NJODL1 liquid culture obtained in Example 2 was centrifuged at 12,000 r / min for 15 minutes, the supernatant was discarded, the bacterial precipitate was collected, and washed 3 times with sterile physiological saline. Then it was inoculated into a 250mL Erlenmeyer flask equipped with 100mL oxalic acid fermentation medium at a volume ratio of 2%, and used as a blank control (with oxalic acid but without bacteria). Bacteria were done in triplicate. Cultivate...

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Abstract

The invention discloses an oxalate-degrading bacterium NJODL1, which is classified as Lactococcus lactis subsp. lactis and has a preservation number of CGMCC NO.4582. The strain has the functions of degrading oxalic acid, regulating oxalic acid metabolism and preventing pet calcium oxalate stones. The oxalate-degrading bacteria NJODL1 has good tolerance to acid and bile salts, no antibiotic resistance to most antibiotics, safe for mice, and can colonize in dogs. NJODL1 can reduce the absorption of oxalic acid in food by the gastrointestinal tract, thereby reducing the content of oxalic acid in urine, thereby reducing the incidence of calcium oxalate stones in dogs. No matter the finished product is food or medicine or yogurt and fermented milk, the strain has good acid resistance and bile salt ability, and has safe enterococcus, which can significantly reduce urine oxalate and prevent pet calcium oxalate stones, and has a wide range of applications prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an oxalic acid degrading bacterium NJODL1 and its application, in particular to the bacterial strain NJODL1 capable of degrading oxalic acid, regulating oxalic acid metabolism and preventing calcium oxalate calculus and its application in the production of pet food, food additives or medicines . Background technique [0002] The incidence of urolithiasis in the global population ranges from 6% to 10%. And about 70% to 80% of urinary stones are calcium oxalate stones. And so far, due to the low solubility of calcium oxalate, the method for treating calcium oxalate stones is still mainly surgical removal, which brings great pain to patients. Some studies have reported that in the formation of calcium oxalate stones, the abnormal absorption and metabolism of oxalic acid play a greater role than calcium, and its slight changes can lead to the generation of calcium oxalate crystals. The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/16A23L1/30A23C9/12A61K35/74A61P13/04C12R1/46A23K10/18A23L33/135A61K35/744
Inventor 黄克和任志华
Owner NANJING AGRICULTURAL UNIVERSITY
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