A methanotroph with denitrification function and hypoxia resistance and its application
A methane-oxidizing bacteria and stress-resistant technology, applied in the field of microorganisms, can solve the problems that aerobic methane-oxidizing bacteria cannot survive for a long time, and achieve the effects of contributing to sustainable development, reducing accidents, and stabilizing performance
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Embodiment 1
[0041] Methanotrophs Methylobacter Isolation and identification of sp. YRD-M2.
[0042] 1. Enrichment culture of methanotrophs:
[0043] Pre-incubation culture was carried out on the wetland soil of the Yellow River Delta. Take 10 g of fresh soil in a 125 mL vial and seal it with butyl rubber and an aluminum cap. Add about 5% methane (6 mL) as a substrate and culture statically in a dark room at 30 °C. During this period, 0.2 mL of gas in the bottle was taken with a gas sampling needle, and the methane concentration was detected by high-performance gas chromatography (Agilent 7820A, USA). The pre-incubated soil was used as the inoculum for methane oxidation enrichment culture. Inoculate 2.5 g of pre-incubated soil into 25 mL of nitrate mineral salt (NMS) medium. The preparation process of NMS medium is as follows: Weigh 0.2 g MgSO with an electronic balance 4 ·7H 2 O, 0.14 g CaCl 2 ·6H 2 O and 1.0 g KNO 3 , and use a syringe to take 50 mL of phosphate buffer (5.44 ...
Embodiment 2
[0055] Methanotrophs Methylobacter The effect of sp. YRD-M2 utilizing methane and nitrate under different methane concentration, air volume and nitrate concentration.
[0056] 1) Preparation of NMS medium:
[0057] Weigh 0.2 g MgSO with an electronic balance 4 ·7H 2 O, 0.14 g CaCl 2 ·6H 2 O and 1.0 g KNO 3 , and use a syringe to take 50 mL of phosphate buffer (5.44 g / L KH 2 PO 4 , 5.68 g / L Na 2 HPO 4 ) and 2 mL trace element solution (1.0 g Na per liter 2 -EDTA, 2.0 g FeSO 4 ·7H 2 O, 0.8 g ZnSO 4 ·7H 2 O, 0.03 g MnCl 2 4H 2 O, 0.03g H 3 BO 3 , 0.2 g CoCl 2 ·6H 2 O, 0.6 g CuCl 2 2H 2 O, 0.02 g NiCl 2 ·6H 2 O and 0.05 g Na 2 MoO 4 2H 2 O), add it to deionized water, mix well and adjust the volume to 1 L, add sodium hydroxide to adjust the pH value to 7.0, take 25 ml of the medium and put it into a 125 mL vial, seal it with a butyl rubber stopper, and then use aluminum Cover and seal, and sterilize at 121°C for 20 min to obtain culture medium.
[005...
Embodiment 3
[0065] Methanotrophs Methylobacter Methane and nitrate utilization effects of sp. YRD-M2 after prolonged hypoxic culture.
[0066] 1) Preparation of NMS medium under hypoxic conditions:
[0067] Weigh 0.2 g MgSO with an electronic balance 4 ·7H 2 O, 0.14 g CaCl 2 ·6H 2 O and 0.23 g KNO 3 , and use a syringe to take 50 mL of phosphate buffer (5.44 g / L KH 2 PO 4 , 5.68 g / L Na 2 HPO 4 ) and 2 mL trace element solution (1.0 g Na per liter 2 -EDTA, 2.0 g FeSO 4 ·7H 2 O, 0.8 g ZnSO 4 ·7H 2 O, 0.03 g MnCl 2 4H 2 O, 0.03 g H 3 BO 3 , 0.2 g CoCl 2 ·6H 2 O, 0.6 g CuCl 2 2H 2 O, 0.02 g NiCl 2 ·6H 2 O and 0.05 gNa 2 MoO 4 2H 2 O), add it to deionized water, mix well and adjust the volume to 1 L, add sodium hydroxide to adjust the pH value to 7.0, take 25ml of the medium and put it into a 125mL vial, seal it with a butyl rubber stopper, and then cover it with an aluminum cap seal. Use an intelligent anaerobic preparation instrument (Beijing Asp Technology Co.,...
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