CN110257291B - Achromobacter capable of tolerating nickel ion toxicity and application thereof - Google Patents

Achromobacter capable of tolerating nickel ion toxicity and application thereof Download PDF

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CN110257291B
CN110257291B CN201910552276.3A CN201910552276A CN110257291B CN 110257291 B CN110257291 B CN 110257291B CN 201910552276 A CN201910552276 A CN 201910552276A CN 110257291 B CN110257291 B CN 110257291B
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achromobacter
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陈倩
孙卫玲
王婷
倪晋仁
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    • C—CHEMISTRY; METALLURGY
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Abstract

The invention relates to an Achromobacter sp resistant to nickel ion toxicity and application thereof. The invention provides an Achromobacter sp with nickel ion tolerance, and the preservation number is CGMCC No. 2964. When the concentration of nickel ions in the sewage is within the range of 0-10 mg/L, the bacterial strain can use nitrate nitrogen as a unique nitrogen source, under the aerobic condition, the removal rate of the nitrate nitrogen can reach 100%, and the removal rate of total nitrogen also reaches 100%. The method has the advantages of convenient operation, high denitrification efficiency and the like, and has important significance for optimizing the sewage biological treatment system and weakening the influence of nickel ions on the sewage denitrification system.

Description

Achromobacter capable of tolerating nickel ion toxicity and application thereof
Technical Field
The invention belongs to the field of biological sewage treatment, and particularly relates to a colorless bacillus capable of tolerating nickel ion toxicity under aerobic conditions and synchronously removing nitrate nitrogen in a water body and application thereof in sewage treatment.
Background
With the acceleration of urbanization and industrialization in China and the continuous development of industrial production, a large amount of industrial wastewater is generated, and great threats are generated to human life and health. As the industrial wastewater treatment capacity of China cannot keep pace with the scale of industrial water, part of industrial wastewater is directly discharged into a town sewage pipe network without treatment. Nickel ions, an important industrial material, are widely used in chemical, metallurgical, electroplating, tanning and coating industries, which produce a large amount of nickel-containing wastewater, and the wastewater inevitably enters municipal sewage treatment plants. Studies have shown that 0.5mg/L Ni (II) negatively affects microbial growth, extracellular polymer formation and other catabolic processes.
In recent years, the discovery of aerobic denitrification (i.e., the process of reducing nitrate nitrogen to nitrogen-containing gas under aerobic conditions) has provided a theoretical basis for achieving simultaneous nitrification and denitrification in a single aerobic reactor. Since the first aerobic denitrifying bacteria T.pantotropia were successfully isolated, more efficient aerobic denitrifying bacteria were isolated in recent years and applied to practical sewage treatment systems. As a novel biological denitrification technology, aerobic denitrification is rapidly developed in recent years due to the advantages of simple process, good denitrification effect, no need of adding acid and alkali and the like.
Nickel (Ni) is a common heavy metal, and is one of the important constituents of earth crust, accounting for about 2% of the total weight. Nickel is present in water bodies generally as ni (ii), occasionally as ni (iii), and ni (iii) can be converted intracellularly (anaerobically) to ni (ii). Ni (II) is an ion necessary for the growth and metabolism of certain microbial cells and is present in an activator of hydrolase (e.g., coenzyme F)430) Among them. Trace amount of Ni (II) can promote the growth of microbes, but Ni (II) also has strong acute biotoxicity, and can produce toxic effect on microbes when the concentration of Ni (II) exceeds a certain concentration.
Aiming at the problem that nickel-containing wastewater enters domestic sewage and Ni (II) and nitrate nitrogen coexist, how to effectively avoid the influence of nickel ions on the removal of the nitrate nitrogen becomes the focus of attention of people, therefore, the development of aerobic denitrifying bacteria with nickel ion toxicity resistance and the application of the aerobic denitrifying bacteria in sewage treatment can effectively cut down the potential risk of the nickel ions on the biological denitrification process of the sewage, and have important significance for ensuring the normal operation of a town sewage treatment system.
Disclosure of Invention
The invention aims to provide a high-efficiency aerobic denitrification strain capable of resisting nickel ion toxicity, which can be used for enhancing the biological denitrification effect of a sewage treatment system so as to reduce the influence of the existence of high-concentration nickel ions on the biological denitrification process of sewage treatment.
The Achromobacter sp is an aerobic denitrifying bacterium capable of removing nitrate nitrogen in a single aerobic environment.
The invention is realized by the following technical scheme:
the Achromobacter sp with nickel ion tolerance provided by the invention has the preservation number of CGMCC No. 2964.
Achromobacter sp as described above, characterized in that: under the condition of different nickel ion concentration levels, the Achromobacter sp can still keep an intact cell structure, and the cell membrane is not damaged.
Achromobacter sp as described above, characterized in that: the strain can perform aerobic denitrification by taking nitrate nitrogen as a nitrogen source and organic matters as a carbon source under the condition of high-concentration nickel ions, so that the nitrate nitrogen is removed.
Use of Achromobacter sp as defined above in the treatment of wastewater, characterized in that: achromobacter sp is added into the nickel-containing sewage, and a proper amount of carbon source is added for aeration, so that the nitrate nitrogen in the sewage can be removed.
The method described above, characterized in that: controlling the temperature in the nickel-containing sewage to be 30 ℃, the pH value to be 7.5, the dissolved oxygen to be 6mg/L and the initial C/N ratio to be 4.
The method described above, characterized in that: when the concentration range of nickel ions in the sewage is 0-10 mg/L, the removal rate of nitrate nitrogen can reach 100%, and the removal rate of total nitrogen can also reach 100%.
The invention has the beneficial effects that:
(1) the Achromobacter sp 1 can perform aerobic denitrification by taking nitrate nitrogen as a nitrogen source and organic matters as a carbon source under the condition of high-concentration nickel ions, so that the nitrate nitrogen is efficiently removed; the problem that anoxic denitrification and aerobic nitrification are required to be adopted for segmented treatment in the conventional wastewater treatment for biological denitrification is solved; in addition, the process flow is simplified, and the cost of equipment and investment is saved, so that the method has great economic benefit and environmental protection benefit;
(2) the strain is inoculated to the biological treatment of the nickel-containing sewage, under the condition of 0-10 mg/L Ni (II), preferably 0-5 mg/L Ni (II), the removal rate of nitrate nitrogen within 20h can reach 100%, and the strain is proved to have stronger nickel ion toxicity resistance and aerobic denitrification capability. The practicability of the strain in urban sewage treatment systems is greatly enhanced due to the characteristics.
Drawings
FIG. 1 shows aerobic denitrification characteristics of Achromobacter in the presence of 0mg/L Ni (II).
FIG. 2 shows aerobic denitrification characteristics of Achromobacter in the presence of 2mg/L Ni (II).
FIG. 3 shows aerobic denitrification characteristics of Achromobacter in the presence of 5mg/L Ni (II).
FIG. 4 shows aerobic denitrification characteristics of Achromobacter in the presence of 10mg/L Ni (II).
Detailed Description
The present invention will be further described with reference to the following examples, but the present invention is not limited to the following examples.
In the following examples, unless otherwise specified, all methods are conventional.
In the following examples, the percentages are by mass unless otherwise specified.
EXAMPLE 1 aerobic Denitrification of Achromobacter in the Presence of 0mg/L Ni (II)
Denitrification Performance testThe formula of the culture medium (BM) is as follows: 8.45g CH3COONa,0.63g NH4Cl,0.61g NaNO3,1.76g K2HPO4·3H2O,0.20g MgSO4·7H2O,0.02g CaCl2,0.005g FeSO4·7H2O,0.1mL of trace element solution. The pH of the medium was adjusted to 7.5 and sterilized at 121 ℃ for 30 min.
Inoculating Achromobacter sp to BM culture medium, performing shake culture at 30 deg.C and 150rpm, and sampling 100 μ L headspace gas with valve needle every 5h for measuring N2O, 2mL of gas was drawn with a sterile syringe and injected into a 2L pure helium bag for NO determination. Meanwhile, 2mL of bacterial suspension is extracted by using a sterile syringe, the bacterial liquid is centrifuged for 5min at 8000rpm at 4 ℃, and the supernatant is taken for analyzing the concentrations of ammonia nitrogen, nitrate nitrogen and nitrite nitrogen.
As shown in FIG. 2, 100mg/L nitrate nitrogen was used by the strain immediately after inoculation and was completely consumed within 10h, with an average removal rate of nitrate nitrogen of 10.0 mg/L/h. Nitrite nitrogen accumulated rapidly with reduction of nitrate nitrogen and reached a maximum of 30.2mg/L over 5h and then was reduced within 10 h. N is a radical of2O also gradually accumulated to a maximum of 0.84mg/L over 10h and then was completely reduced over 15 h. The highest value of the accumulation amount of the intermediate product NO in the denitrification process is 7.1 mu g/L, and the accumulation amount is less, which indicates that the strain has better aerobic denitrification capability.
EXAMPLE 2 aerobic denitrification Properties of Achromobacter in the Presence of 2mg/L Ni (II)
The denitrification performance test medium (BM) formulation is as follows: 8.45g CH3COONa,0.63g NH4Cl,0.61g NaNO3,1.76g K2HPO4·3H2O,0.20g MgSO4·7H2O,0.02g CaCl2,0.005g FeSO4·7H2O,0.1mL of trace element solution. The pH of the medium was adjusted to 7.5 and sterilized at 121 ℃ for 30 min.
Inoculating Achromobacter sp to BM medium containing 2mg/L Ni (II), culturing at 30 deg.C and 150rpm with shaking, and feeding with valve every 5hThe sample needle extracts 100. mu.L headspace gas for measuring N2O, 2mL of gas was drawn with a sterile syringe and injected into a 2L pure helium bag for NO determination. Meanwhile, 2mL of bacterial suspension is extracted by using a sterile syringe, the bacterial liquid is centrifuged for 5min at 8000rpm at 4 ℃, and the supernatant is taken for analyzing the concentrations of ammonia nitrogen, nitrate nitrogen and nitrite nitrogen.
As shown in FIG. 2, 100mg/L nitrate nitrogen was used by the strain immediately after inoculation and was completely consumed within 20h, with an average removal rate of nitrate nitrogen of 10.0 mg/L/h. Nitrite nitrogen accumulated rapidly with the reduction of nitrate nitrogen and reached a maximum of 32.1mg/L over 10h and then was reduced over 20 h. N is a radical of2O also gradually accumulated to a maximum of 1.15mg/L over 15h and then was completely reduced over 20 h. The highest value of the accumulation amount of the intermediate product NO in the denitrification process is 7.5 mu g/L, and the accumulation amount is less, which shows that the strain can still better exert the aerobic denitrification capability in the presence of 2mg/L Ni (II).
EXAMPLE 3 aerobic denitrification Properties of Achromobacter in the Presence of 5mg/L Ni (II)
Achromobacter sp was inoculated into BM medium containing 5mg/L of Ni (II) to test aerobic denitrification performance. As shown in FIG. 3, 100mg/L nitrate nitrogen was used by the strain immediately after inoculation and was completely consumed within 20h, with an average removal rate of 5.0 mg/L/h. Nitrite nitrogen gradually accumulated with the reduction of nitrate nitrogen and reached a maximum of 32.2mg/L over 15h and then was reduced over 30 h. N is a radical of2O also gradually accumulated to a maximum of 2.63mg/L over 20h and then was completely reduced over 30 h. It must be noted that the maximum value of the NO accumulation amount of the intermediate product in the denitrification process at this time is 4.2. mu.g/L, and only accounts for 0.004% of the nitrate nitrogen removal amount. Therefore, the strain still has high-efficiency aerobic denitrification capability under the condition of 5mg/L Ni (II).
EXAMPLE 4 aerobic denitrification Properties of Achromobacter in the Presence of 10mg/L Ni (II)
Achromobacter sp was inoculated into BM medium containing 10mg/L of Ni (II) to test aerobic denitrification performance. The results are shown in FIG. 4, 100mg/L nitrate nitrogenImmediately after inoculation, it was used by the strain and completely consumed within 25h, with an average nitrate nitrogen removal rate of 4.0 mg/L/h. Nitrite nitrogen gradually accumulated with the reduction of nitrate nitrogen and reached a maximum of 38.3mg/L over 20h and then was reduced over 35 h. At the same time, N2O also gradually accumulated to a maximum of 3.42mg/L over 25h and was then completely reduced over 40 h. It should be noted that the maximum value of the accumulated NO amount of the intermediate product in the denitrification process is 6.4. mu.g/L, and only accounts for 0.006% of the nitrate nitrogen removal amount. Therefore, the strain still has high-efficiency aerobic denitrification capability under the condition of 10mg/L Ni (II).
EXAMPLE 5 aerobic Denitrification of Achromobacter in the Presence of 50mg/L Ni (II)
Achromobacter sp was inoculated into BM medium containing 50mg/L of Ni (II) to test aerobic denitrification performance. The growth of the strain was significantly inhibited and nitrate nitrogen was hardly removed, and it can be seen that 50mg/L Ni (II) is the upper concentration limit of the strain, and the strain does not have aerobic denitrification capability under this condition. However, it can be seen from the above examples 1-4 that when the strain is inoculated into the biological treatment of nickel-containing sewage, the removal rate of nitrate nitrogen can reach 100% in the presence of 0-10 mg/L Ni (II), which indicates that the strain has strong nickel ion toxicity resistance and aerobic denitrification capability (see Table 1).
TABLE 1 Denitrification Properties of Achromobacter in the Presence of different concentrations of Ni (II)
Ni(II)(mg/L) 25h nitrate nitrogen removal (%) 35h Total Nitrogen removal (%)
2 100％ 100％
5 100％ 100％
10 100％ 100％
Example 6 aerobic Denitrification Properties of Achromobacter and Pseudomonas stutzeri in the Presence of 10mg/L Ni (II)
Achromobacter (Achromobacter sp.) and Pseudomonas stutzeri (P.stutzeri) were inoculated into BM medium containing 10mg/L of Ni (II) to test aerobic denitrification performance. As a result, as shown in Table 2, 100mg/L of nitrate nitrogen was available to Achromobacter immediately after inoculation, the maximum removal rate of nitrate nitrogen was 6.3. + -. 0.3mg/L/h, and the final product was N2Indicating that the denitrification process is not inhibited. And under the condition that the Pseudomonas stutzeri exists in 10mg/L of Ni (II), the maximum removal rate of nitrate nitrogen is only 2.0 +/-0.3 mg/L/h, and the final denitrification product is nitrite nitrogen, which indicates that the denitrification process is not completely carried out. As is known, Pseudomonas stutzeri is a typical denitrifying strain, and by comparison, the Achromobacter provided by the invention can have high tolerance to Ni (II) while performing efficient aerobic denitrification (see Table 2).
TABLE 2 comparison of aerobic denitrification Properties of Achromobacter and Pseudomonas stutzeri in the Presence of 10mg/L Ni (II)
Figure BDA0002105823170000051
Example 7 application of Achromobacter in Nickel-containing wastewater
Adding achromobacter into an activated sludge system to treat nickel-containing wastewater, continuously aerating to keep the dissolved oxygen of the system at 6mg/L, and treating the wastewater with the following water quality: pH 7.5, nitrate nitrogen content 100mg/L, Ni (II)10 mg/L. The results are shown in table 2, the total nitrogen removal rate under aerobic conditions is lower in the common activated sludge system; the aerobic denitrification performance was affected by the addition of 10mg/L Ni (II). However, the nitrate nitrogen removal at 36h was 96.4% and the total nitrogen removal was 90.5% for the achromobacter enhanced system (see table 3).
TABLE 3 biological denitrification results of Achromobacter in activated sludge systems
Figure BDA0002105823170000061
Therefore, aiming at the problem that the nickel-containing wastewater enters the domestic sewage and Ni (II) and nitrate nitrogen coexist, the strain can effectively avoid the influence of nickel ions on the removal of the nitrate nitrogen, can be widely applied to sewage treatment, can effectively reduce the potential risk of the nickel ions on the biological denitrification process of the sewage, and has important significance for ensuring the normal operation of the urban sewage treatment system.
It should be noted that the above-mentioned embodiments are only for describing the invention in further detail, and are not intended to limit the invention, and those skilled in the art can make various modifications or changes within the scope not departing from the spirit and spirit of the invention, and still fall into the protection scope of the appended claims.

Claims (2)

1. Achromobacter (A) capable of resisting nickel ion toxicityAchromobacter sp.) The application of (2), which is characterized in that: the application is that aerobic denitrifying bacteria Achromobacter (Achromobacter) is usedAchromobacter sp.) The biological denitrification effect of the sewage treatment system is enhanced, and the influence of the existence of excessive nickel ions on the biological denitrification process of the sewage treatment is reduced; the preservation number of the achromobacter is CGMCC No.2964, and the achromobacter has better tolerance capability to nickel ions with different concentration levels; the application specifically comprises the following steps: adding the colorless rod into the nickel-containing sewageBacteria (A), (B)Achromobacter sp.) And adding a proper amount of carbon source for aeration, so that the nitrate nitrogen in the sewage can be removed; the temperature of the nickel-containing sewage is 30 ℃; the pH value of the nickel-containing sewage is 7.5; the dissolved oxygen in the nickel-containing sewage is 6 mg/L; the initial C/N ratio in the nickel-containing wastewater is 4.
2. Use according to claim 1, characterized in that: the concentration range of nickel ions contained in the sewage is 0-10 mg/L, the removal rate of nitrate nitrogen can reach 100% under an aerobic environment, and the removal rate of total nitrogen can also reach 100%.
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