Marine nitrobacterium, preparation method therefor and application of marine nitrobacterium
A technology of nitrifying bacteria and oceans, applied in biochemical equipment and methods, chemical instruments and methods, bacteria, etc., can solve the problem of reduced ammonia nitrogen removal capacity and other issues
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Embodiment 1
[0049] Embodiment 1: the preparation method of above-mentioned suspicious sulfite bacteria (Sulfitobacter dubius) PT04 comprises:
[0050] S1. Isolation and purification of suspected sulfite bacteria (Sulfitobacter dubius) PT04 strain:
[0051] S101. Take 1 g of the deep-sea sediment sample, insert it into 100 mL of heterotrophic nitrification liquid medium supplemented with 100 mg / L Cr(VI), and enrich and culture it on a shaking table at 30°C and 170 rpm for 5 days;
[0052] S102. Then transfer to a new enrichment medium at a ratio of 1%, and continue to enrich for 5 days;
[0053] S103. After enrichment, use the dilution coating plate method to dilute in a 10-fold gradient, draw 0.2 mL each, and spread it on a heterotrophic nitrification solid medium containing the same concentration of Cr(VI) and ammonia nitrogen, and culture it upside down at 30°C for 24 hours;
[0054] S104. Observe and record the characteristics of different single bacterial colonies, pick out single ba...
Embodiment 2
[0067] Embodiment 2: For the research of suspicious sulfite bacteria (Sulfitobacter dubius) PT04 applied to the reduction of hexavalent chromium in leather wastewater, specifically to explore pH, temperature and salinity on suspicious sulfite bacteria PT04 ammonia nitrogen degradation and Cr( VI) Effect of reduction:
[0068] S1. Exploring the effect of pH value on ammonia nitrogen degradation and Cr(VI) reduction:
[0069] Different microorganisms have their own optimum pH for growth. If the pH is too high or too low, it will affect their growth and metabolism, which in turn will affect the ability of microorganisms to treat biological substances;
[0070] The initial pH value of the medium was adjusted to 4.0, 5.0, 6.0, 7.0, 8.0 with NaOH and HCl; the concentration of Cr(VI) added in the heterotrophic nitrification liquid medium was 100 mg / L, and the concentration of NaCl was 5%; inoculated with PT04 After culturing at 170rpm and 30°C for 3 days, the effects of different pH...
Embodiment 3
[0077] Example 3: Exploring the reduction principle, biosafety, bioremediation and acute toxic effects on Artemia of suspected sulfite bacteria (Sulfitobacter dubius) PT04:
[0078] S1. Exploring the localization of Cr(VI) reductase and ammonia nitrogen degrading enzyme
[0079]After PT04 was cultured in the heterotrophic nitrification liquid medium for 24 hours, it was centrifuged at 6000g for 10 minutes; the supernatant of the bacterial liquid was stored at 4°C before measuring the enzyme activity. The bacterial pellet was washed twice with refrigerated PBS buffer and resuspended to a final concentration of 0.1 g / mL. The bacterial suspension was sonicated in an ice bath environment. 750W 40% power, 10 seconds of ultrasound, 10 seconds interval of ultrasound for 20min. The obtained cell lysate was centrifuged at 175000g for 30min at 4°C. The cell lysate supernatant was filtered through a 0.22 μm filter. The obtained cell-free filtrate is the source of soluble enzymes, whi...
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