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

Active Publication Date: 2021-01-08
MINJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned problems, the present invention aims to provide a marine nitrifying bacterium and its preparation method and application, by providing a highly tolerant to salt content and capable of removing hexavalent chromium and ammonia nitrogen in polluted water sources under aerobic conditions. Sulfitobacter dubius PT04 treats leather wastewater, which solves the problem that the existing nitrifying bacteria are inhibited by toxic substances in leather wastewater, and the ability to remove ammonia nitrogen is reduced. Biological safety, using enzymatic reaction to convert Cr(VI) and ammonia nitrogen into less toxic products, showing good results in the treatment of ammonia nitrogen and hexavalent chromium in leather wastewater, with good reduction effect, high nitrification efficiency, green The characteristics of environmental protection have good application prospects in the field of bioremediation

Method used

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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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Abstract

The invention discloses a marine nitrobacterium, a preparation method therefor and application of the marine nitrobacterium. Through treating leather wastewater with sulfitobacter dubius PT04, which has high saline matter tolerance and can remove hexavalent chrome and ammonia nitrogen from a contaminated water source under aerobic conditions, the problem that the hexavalent chrome and ammonia nitrogen removing capability of the existing nitrobacteria is lowered due to inhibition of toxic substances in the leather wastewater is solved; and through experiments, it is proven that the sulfitobacter dubius PT04 has relatively high biological safety, the Cr(VI) and the ammonia nitrogen are converted into products with lower toxicity by using an enzymatic reaction, and the marine nitrobacterium shows a good effect in treatment on the ammonia nitrogen and hexavalent chrome in the leather wastewater, has the advantages of good reduction effect, high nitrifying efficiency and environment-friendliness and has a good application prospect in the field of bioremediation.

Description

technical field [0001] The invention relates to the technical field of leather wastewater treatment, in particular to a marine nitrifying bacterium and its preparation method and application. Background technique [0002] Leather is an important pillar industry of my country's light industry, but the pollution it brings has also caused damage to the environment. Leather wastewater has the characteristics of high salinity, high ammonia nitrogen, low nutrition, and high heavy metal chromium content. Among them, the chromium-containing wastewater mainly comes from the chrome tanning process. , since the toxicity of hexavalent chromium is 100 times that of trivalent chromium, the usual method to deal with hexavalent chromium pollution is to convert hexavalent chromium into trivalent chromium. , low operating costs, no secondary pollution and other advantages have attracted more and more attention; [0003] The input of high-ammonia-nitrogen wastewater causes lake eutrophication a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/01C02F101/22C02F103/24
CPCC12N1/20C12N1/02C02F3/34C02F2101/22C02F2103/24C12R2001/01C12N1/205
Inventor 王蔚林镇跃梁晨潘长坤赖庆娜陈建明
Owner MINJIANG UNIV
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