Oxalate-degrading strain NJODE1 and application thereof
A technology for degrading bacteria and oxalic acid, which is applied in the biological field to achieve the effects of strong oxalic acid degradation ability, excellent acid resistance and bile salt resistance, and no antibiotic resistance.
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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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