A method for separating non-peroxidase enzyme components capable of degrading zearalenone toxin
A zearalenone, non-peroxidase technology, applied in the field of microbial application, can solve the problem of non-peroxidase purification and the like
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Embodiment 1
[0072] (1) Culture: Inoculate with 1% (v / v) Acinetobacter sp.SM04 suspension (OD 600 =0.6) culture (M2 medium) was cultured in an air-bath shaker at 28°C (150r / min) for 24h, the liquid culture was centrifuged for 10min (8000×g, 4°C), and the supernatant after centrifugation was used 0.22μm membrane filtration, and the filtrate was concentrated 8 times into a crude enzyme solution at 45°C using a vacuum rotary evaporator;
[0073] The formulation of the medium used is as follows:
[0074] M2 medium: 12.5g sodium acetate, 2.5g NH 4 NO 3 , 1.2gK 2 HPO 4 ·3H 2 O, 1gKCl, 0.4gMgSO 4 ·7H 2 O, add 10mL trace element stock solution, add distilled water to make up to 1000mL, adjust the pH to 7.0 after mixing;
[0075] The formula of described trace element stock solution is as follows: 2g / LFeSO 4 ·7H 2 O, 0.5g / LMnSO 4 4H 2 O, 0.4g / LCuSO 4 ·5H 2 O, 0.5g / LCoCl 6 ·6H 2 O and 0.4g / LZnCl 2 .
[0076] (2) Add the crude enzyme solution in step (1) to an anionic Sephadex colum...
Embodiment 2
[0084] (1) Culture: Inoculate with 2% (v / v) Acinetobacter sp.SM04 suspension (OD 600 =0.8) culture (M2 medium) was cultured in an air-bath shaker at 30°C (180r / min) for 36h to the end of logarithmic growth, and the liquid culture was centrifuged for 12min (9000×g, 5°C). The supernatant was filtered with a 0.22 μm filter membrane, and the filtrate was concentrated 9 times at 50°C using a vacuum rotary evaporator to become a crude enzyme solution;
[0085] The formulation of the medium used is as follows:
[0086] M2 medium: 15g sodium acetate, 2.75g NH 4 NO 3 , 1.35gK 2 HPO 4 ·3H 2 O, 1.25gKCl, 0.5gMgSO 4 ·7H 2 O, add 10mL trace element stock solution, add distilled water to make up to 1000mL, adjust the pH to 7.3 after mixing;
[0087] The formula of the trace element stock solution is the same as that in Example 1-(1).
[0088] (2) Add the crude enzyme solution in step (1) to an anionic Sephadex column DEAESephadexA-50 column (2.0cm×15cm), and use sodium phosphate sa...
Embodiment 3
[0094] (1) Culture: Inoculate with 3% (v / v) Acinetobactersp.SM04 bacterial suspension (OD 600 =1.0) culture (M2 medium) was cultured in an air bath shaker at 28°C (200r / min) for 48h, the liquid culture was centrifuged for 15min (10000×g, 6°C), and the supernatant after centrifugation was used 0.22μm membrane filtration, and the filtrate was concentrated 10 times by a vacuum rotary evaporator at 55°C to form a crude enzyme solution;
[0095] The formulation of the medium used is as follows:
[0096] M2 medium: 17.5g sodium acetate, 3g NH 4 NO 3 , 1.5gK 2 HPO 4 ·3H 2 O, 1.5gKCl, 0.6gMgSO 4 ·7H 2 O, add 10mL trace element stock solution, add distilled water to make up to 1000mL, adjust the pH to 7.5 after mixing;
[0097] The formula of the trace element stock solution is the same as that in Example 1-(1).
[0098] (2) Add the crude enzyme solution in step (1) to an anionic Sephadex column DEAESephadexA-50 column (2.0cm×15cm), and use sodium phosphate salt with pH 7.0, 6...
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