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Oxalate-degrading strain NJODE1 and application thereof

A technology for degrading bacteria and oxalic acid, applied in the biological field, to achieve the effects of no antibiotic resistance, excellent acid resistance and bile salt resistance, and strong ability to degrade oxalic acid

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

[0009] Therefore, isolating probiotic enterococci that can degrade 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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  • Oxalate-degrading strain NJODE1 and application thereof
  • Oxalate-degrading strain NJODE1 and application thereof
  • Oxalate-degrading strain NJODE1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Isolation and identification of Enterococcus faecium

[0042] 1. Preparation of culture medium

[0043] The formula of Pfizer Enterococcus selective solid medium is: 1L distilled water contains: 20.0g peptone, 5.0g yeast extract, 10.0g bovine bile, 5.0g sodium chloride, 1.0g sodium citrate, 0.5g ferric ammonium citrate , sodium azide 0.25g, escin 1.0g, agar 15.0g, adjust pH=7.0-7.3; 121°C, autoclave for 15min, set aside.

[0044] 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 80 1mL, MgSO 4 ·7H 2 O 0.58g, MnSO 4 4H 2 O 0.05g, 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.

[0045] 2. Sample source

[0046] 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...

Embodiment 2

[0061] Embodiment 2: the bacterial strain NJODE1 screening of embodiment 1 expands culture

[0062] 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 80 1mL, MgSO 4 ·7H 2 O 0.58g, MnSO 4 4H 2 O 0.05g, 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.

[0063] The bacterial strain NJODE1 obtained in Example 1 was inoculated in a 250mL Erlenmeyer flask containing 50mL of seed medium, and cultivated for 24h at a temperature of 37°C to obtain a NJODE1 liquid culture with a cell count of 3.5×10 9 CFU.mL -1 .

Embodiment 3

[0064] Embodiment 3: Use bacterial strain NJODE1 to ferment and degrade oxalic acid in the MRS fermentation medium containing 20mmol / L oxalic acid

[0065] 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 80 1mL, MgSO 4 ·7H 2 O 0.58g, MnSO 4 4H 2 O 0.05g, 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.

[0066]The NJODE1 liquid culture obtained in Example 2 was centrifuged at 12000r / min for 15 minutes, the supernatant was discarded, and 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 5%, and used as a blank control (with oxalic acid but without bacteria). Bacteria were made in triplicate. Cultivate...

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Abstract

The invention discloses an oxalate-degrading strain NJODE1 and application thereof. The oxalate-degrading strain NJODE1 has a classification name of Enterococcus faecium with the preservation number of CGMCC NO. 4581. The oxalate-degrading strain NJODE1 has good tolerance to acids and bile salts, has no antibiotic resistance to most non-antibiotics, is safe for mice and can be colonized in dogs. The NJODE1 can reduce gastrointestinal absorption of oxalate in foods, thereby reducing the content of oxalate in urine, and thus reducing the incidence of canine calcium oxalate calculus. Whether thefinished product of the strain is food or pharmaceutical products or yogurt and fermented milk, the strain has good resistance to acids and bile salts, contains safe enterococci, and has significant effects on lowering the urinary oxalate content and preventing and treating calcium oxalate calculus of pets, thus having broad application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an oxalic acid degrading bacterium NJODE1 and its application, in particular to a bacterial strain NJODE1 capable of degrading oxalic acid, regulating oxalic acid metabolism and preventing calcium oxalate stones, and its application in pet food, food additives or medicines application in production. 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. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/16A23L1/30A23C9/12A61K35/74A61P13/04C12R1/46A23K10/18A23L33/135A61K35/741
Inventor 黄克和任志华蒋鲁岩陈兴祥
Owner NANJING AGRICULTURAL UNIVERSITY
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