CN110283739A - The denitrifying bacteria of one plant of salt tolerant and its application - Google Patents
The denitrifying bacteria of one plant of salt tolerant and its application Download PDFInfo
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Abstract
The denitrifying bacteria of one plant of salt tolerant and its application, the present invention relates to technical field of environmental microorganism.The technical issues of denitrogenating ability the invention solves existing common denitrifying bacteria to be inhibited by high salinity, saliferous breeding wastewater Nitrite Nitrogen can not being removed.The bacterial strain is other style Shewanella Alishewanella sp.F2, it is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the deposit date is on March 25th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, and deposit number is CGMCC No:17433.The bacterial strain can denitrogenation under the conditions of saliferous, carry out denitrification.Bacterial strain of the present invention is for removing Nitrite nitrogen pollutant.
Description
Technical field
The present invention relates to technical field of environmental microorganism.
Background technique
With the rapid development of sea-farming, it is most by enter that half is intensive or intensive culture in the form of, breeding density mistake
High and feeding residual bait excessively waits factors to cause breeding water body Nitrite Nitrogen and increases year by year.Nitrite nitrogen is not only
The intermediate product of one of water eutrophication factor and nitrification and denitrification, therefore it is widely present in Natural Water
In body, high risks easily are caused to aquatic animal and human health.Nitrite toxicity is big, and the mankind drink for a long time
With high concentration nitrous acid salt water, feeblemindedness will lead to, or even dead；Aquatic animal is due to it accumulates nitrite to nitrous acid
Toxicity is more sensitive, causes a variety of physiology disorder such as aquatic animal tissue oxygen content is insufficient, be short of breath, palpitating speed existing
As.Therefore, it reduces or removal Nitrite is particularly significant safely to water body.
The method of removal Nitrite nitrogen includes physical method, chemical method and biological method, biological method because
It can effectively remove a variety of nitrogenous compounds and secondary pollution is not brought to obtain extensive research, be most effective and green ring
One of method of guarantor.Nitrite nitrogen in microbial process removal water body is mainly the nitrification and denitrification work for passing through microorganism
For what is realized, nitrification is the effect of nitrobacteria (nitrite-oxidizing bacteria) using nitrite nitrogen as electron donor
Under be oxidized process for nitrate nitrogen；Denitrification is using nitrite nitrogen as electron acceptor, in denitrifying bacteria
It is reduced to nitrogen oxide or nitrogen under effect, and realizes the process for completely removing Nitrite nitrogen.Denitrification
Effect can fundamentally solve the problems, such as that Water compound is exceeded, it is considered to be the most effective side of denitrogenating in biological denitrificaion
Formula.However, the waste water generated during sea-farming contains great amount of soluble salt, osmotic pressure is caused to increase, easily caused general
The serious dehydration of microbial bacteria body cell, protoplasm separation and cause growth metabolism to be suppressed, or even it is dead.Common denitrification is thin
Bacterium denitrogenates ability and is inhibited by high salinity, can not be removed to saliferous breeding wastewater Nitrite Nitrogen.Therefore, micro- life is utilized
Object denitrogenation processing saliferous breeding wastewater Nitrite Nitrogen becomes research hotspot and difficult point in this field.
Summary of the invention
Ability is denitrogenated the invention solves existing common denitrifying bacteria to be inhibited by high salinity, saliferous can not be cultivated useless
The technical issues of water Nitrite Nitrogen is removed, and the denitrifying bacteria and its application of one plant of salt tolerant are provided.
The denitrifying bacteria of one plant of salt tolerant, the bacterial strain are other style Shewanella Alishewanella sp.F2, are deposited in
State's Microbiological Culture Collection administration committee common micro-organisms center, preservation address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing Cities 3
Number, the deposit date is on March 25th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, compiles
Number be CGMCC No:17433.
Other style Shewanella Alishewanella sp.F2 is by 16S rDNA sequence alignment analysis, with other style Shiva
The homology of the 16S rDNA sequence of Salmonella Alishewanella sp. is 99.81%.By combining morphological features, life
Elongate member, Physiology and biochemistry qualification result determine that bacterial strain F2 of the present invention belongs to other style Shewanella Alishewanella sp. bacterium
Belong to.
The application of the denitrifying bacteria of salt tolerant of the invention denitrogenation under the conditions of saliferous.
Further, the saliferous condition is NaCl concentration≤30g/L.
Further, the saliferous condition pH is 9~10, NaNO2Initial concentration is 0.4~0.8g/L.
Further, the inoculum concentration of the denitrifying bacteria of the salt tolerant is 5~7% (v/v).
The beneficial effects of the present invention are:
The present invention filters out one plant of other style Shewanella (Alishewanella sp.), it is found that it has efficiently removal sub-
The denitrifying capacity of nitrate nitrogen；It is further discovered that this bacterium has certain salt tolerance, is effectively played in saliferous condition
Its denitrifying capacity.It, can be compared with using the physio-biochemical characteristics and metabolic mechanism of this kind of bacterium with salt tolerant denitrifying capacity
Problem present in the common denitrifying bacteria processing saliferous breeding wastewater of good solution.
Alishewanella sp.F2 provided by the invention is one plant of denitrifying bacteria, when inoculum concentration is 5% (v/v),
Its denitrification capability is cultivating 36h clearly, and 48h is to NO2 --N、NO3 -- N removal rate is respectively 98.97% He
94.81%, 120h is to NO2 --N、NO3 -- N removal rate is up to 96% or more.It can be seen that Alishewanella sp.F2 has
Efficient denitrification capability.In addition, there is certain salt tolerant by Salt resistant test result judgement Alishewanella sp.F2
Property, but to will lead to its speed of growth slow for too high in salinity.Under 0~30g/L NaCl Variation of Salinity Condition, Alishewanella
Sp.F2 has efficient NO2 --N、NO3 -- N removal ability, removal rate is 99% or so.
Therefore strains A lishewanella sp.F2 of the present invention handles high concentration nitrite under the conditions of certain saliferous
Waste water has good practical value and application prospect.
The denitrifying bacteria of one plant of salt tolerant, the bacterial strain are other style Shewanella Alishewanella sp.F2, are deposited in
State's Microbiological Culture Collection administration committee common micro-organisms center, preservation address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing Cities 3
Number, the deposit date is on March 25th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, compiles
Number be CGMCC No:17433.
Bacterial strain of the present invention is for removing Nitrite nitrogen pollutant.
Detailed description of the invention
Fig. 1 is that the gas generation phenomenon in other style Shewanella Alishewanella sp.F2 incubation of the present invention is shone
Piece；
Fig. 2 is the pcr amplification product electrophoresis of other style Shewanella Alishewanella sp.F2 genomic DNA of the present invention
Map；F2 is strains A lishewanella sp.F2 of the present invention；
Fig. 3 is the 16S rDNA gene sequence of other style Shewanella Alishewanella sp.F2 of the present invention and similar strain
Arrange the phylogenetic tree spectrogram of building；Wherein, F2 is strains A lishewanella sp.F2 of the present invention；
Fig. 4 is one other style Shewanella Alishewanella sp.F2 of embodiment to NO2 -The removal effect of-N, whereinRepresent NO2 -- N concentration, ■ represent removal rate；
Fig. 5 is one other style Shewanella Alishewanella sp.F2 of embodiment to NO3 -The removal effect of-N, whereinRepresent NO3 -- N concentration, ■ represent removal rate；
Fig. 6 is NO in denitrification culture medium under one different salinity of embodiment2 -- N terminates concentration map, whereinRepresent NO2 --
N concentration, ■ represent removal rate；
Fig. 7 is NO in denitrification culture medium under one different salinity of embodiment3 -- N terminates concentration map, whereinIt represents
NO3 -- N concentration, ■ represent removal rate.
Specific embodiment
Technical solution of the present invention is not limited to the specific embodiment of act set forth below, further include each specific embodiment it
Between any combination.
Specific embodiment 1: the denitrifying bacteria of one plant of salt tolerant of present embodiment, which is other style Shewanella
Alishewanella sp.F2 is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address
It is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the deposit date is on March 25th, 2019 in Chinese microorganism strain preservation pipe
The common micro-organisms center preservation of the reason committee, number are CGMCC No:17433.
Present embodiment other style Shewanella Alishewanella sp.F2 bacterium colony is white, round, diameter be 1~
2mm, neat in edge, surface is smooth, sticky, glossy.Microscopically observation thallus be it is rod-shaped, length is 1~2 μm, and width is
0.3~0.5 μm.
The physiological and biochemical property of present embodiment other style Shewanella Alishewanella sp.F2, as shown in table 1:
The physiological and biochemical property of 1 Alishewanella sp.F2 of table
+: growth is positive；: it does not grow or negative.
Specific embodiment 2: the screening technique of present embodiment other style Shewanella Alishewanella sp.F2 is pressed
Following steps are realized:
Step 1: it draws 2mL and picks up from Zhuanghe City, Daliang City, Liaoning Province (39 ° 38 ' 31 of north latitude " 122 ° 58 ' 19 of east longitude ") sea wall
Muddy water sample be put into 250mL conical flask, the bacterial strain denitrification culture medium of 200mL is added；Using sealed membrane by bottle sealing
Processing manufactures anaerobic environment；Enrichment culture liquid is obtained after constant temperature stationary culture 5~7 days under the conditions of temperature is 30 DEG C；
Step 2: step 1 enrichment culture liquid is carried out by gradient dilution using coubling dilution, draws each dilution respectively
Suspension 0.5mL will be enriched with into bacterial strain screening plating medium (having stood overnight, no varied bacteria growing) with glass triangle spreading rod
After culture solution coating uniformly, the screening flat board culture medium that the sterilized temperature of the second layer is not higher than 40 DEG C is poured into, manufactures anoxic ring
Border is for cultivating denitrifying bacteria；It is using sealed membrane that plate is sealed around；Plate is inverted after the solidification of second layer culture medium,
Being put into temperature is in 30 DEG C of constant incubator, and culture is to growing obvious bacterium colony；The bacterial strain that picking is separated, in the double-deck bacterial strain
Repeatedly scribing line purifying, until microscopically observation shows no miscellaneous bacteria, is then seeded to 200mL's on screening flat board culture medium
In denitrification culture medium, in the case where temperature is 30 DEG C of anoxia condition, constant temperature, which is stood, expands culture；
Step 3: taking step 2 to expand the obtained bacterium solution of culture, by inoculum concentration to be that 5% (v/v) is inoculated into 200mL fresh
In bacterial strain denitrification culture medium, 5d is cultivated in the case where temperature is 30 DEG C of anoxia condition, culture medium cloudiness is observed and is training
Whether support has gas generation in the process；As shown in Figure 1, the culture after culture 5d becomes cloudy, and generate different degrees of bubble；
With NO in bacterial strain denitrification culture medium2 --N、NO3 -The removal rate of-N is screened as index, chooses NO2 --N、NO3 -- N removal
Efficiency reaches 96% or more bacterium solution, through separation, after purification, obtains one plant of bacterial strain F2 with efficient denitrification function, i.e., complete
At the screening of denitrifying bacteria.
Wherein, the bacterial strain denitrification culture medium (/L) is by the CH of 5g3The K of COONa, 1g2HPO4, 0.8g NaNO2、
The CaCl of 0.03g2, 1g Na2CO3, 0.06g FeSO4·7H2O, the MgSO of 0.2g4·7H2O and 1000mL deionized water group
At pH=10, temperature is made of sterilizing under 121 DEG C of condition of high voltage 30min；
Bacterial strain screening plating medium (/L) is every liter of addition 20g agar on the basis of the bacterial strain denitrification culture medium
Composition, pH=10, temperature are made of sterilizing under 121 DEG C of condition of high voltage 30min.
Other style Shewanella Alishewanella sp.F2 is identified:
16S rDNA sequencing and chadogram production:
Bacterial strain F2 genome DNA is extracted: according to a conventional method, through strain culturing, microorganism collection, cellular lysate, DNA mentioned
It takes, precipitates, washing takes out (drying in the air) and does 7 steps, finally obtains genome DNA, 5 μ L is taken to be examined with 1% agarose gel electrophoresis
It surveys.
The PCR amplification of bacterial strain F2 genomic DNA: using total DNA as template, amplimer is a pair of of universal primer.
Forward primer is 27F:5 '-AGAGTTTGATCCTGGCTCAG-3 '；
Reverse primer is 1492R:5 '-GGTTACCTTGTTA CGACTT-3 '.
PCR amplification is carried out by template of genomic DNA.PCR reaction system is as follows:
PCR reaction carries out in 25 μ L systems.The composition of reaction system are as follows: 0.5 μ L of template DNA, 1 μ L of dNTP (mix),
Taq Buffer(with MgCl2) 2.5 μ L, Taq enzyme 0.2 μ L, 0.5 μ L of primers F, 0.5 μ L of primer R plus distilled water be to 25 μ
L。
PCR amplification needs 94 DEG C of initial denaturation 4min, 94 DEG C of denaturation 45s, 55 DEG C of annealing 45s, 72 DEG C of extension 1min, and totally 30
Circulation, 72 DEG C of reparations extend 10min, and 4 DEG C of termination reactions will obtain segment and be used to be sequenced.
The 16S rDNA of other style Shewanella Alishewanella sp.F2 is as shown in SEQ ID NO:1 in sequence table.
Pcr amplification product takes 5 μ L to be detected with 1% agarose gel electrophoresis.
Pcr amplification product electrophorogram is as shown in Figure 2.
Adhesive tape is cut after verifying, is produced with DNA gel QIAquick Gel Extraction Kit (Shanghai Sheng Gong bioengineering Co., Ltd) purifying PCR
Object.
The evolutionary degree of bacterium phylogenetic tree in similar strain is analyzed by N-J algorithm, as a result as shown in figure 3,
The homology of the 16S rDNA sequence of bacterial strain F2 and other style Shewanella Alishewanella sp. is as can be seen from Figure 3
99.81%.
Referring to " common bacteria system identification handbook " and " primary Jie Shi systematic bacteriology handbook ", in conjunction with physiological and biochemical property with
16S rDNA sequencing, identify bacterial strain F2 be other style Shewanella (Alishewanella sp.), 16S rDNA sequence it is homologous
Property is 99.81%.Name bacterial strain F2 are as follows: Alishewanella sp.F2, the deposit date is on March 25th, 2019, preservation address
It is China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No:17433.
Specific embodiment 3: the Salt resistant test of present embodiment other style Shewanella Alishewanella sp.F2,
It is specific as follows:
The screening isolation medium of different salinity gradient processed, i.e. NaCl concentration are respectively 0g/L, 30g/L, 50g/L, 70g/
L,100g/L；By on the culture medium flat plate of bacterial strain F2 streak inoculation to each salinity, it is put into 30 DEG C of constant incubators and cultivates 5d, with
Whether bacterial strain grows the foundation as the whether resistance to corresponding salinity of the bacterial strain；Not to be inoculated with the culture medium of denitrifying bacteria as blank
Control, every group parallel 3.
After 1d is cultivated, each salinity medium does not occur bacterium colony；After 2d is cultivated, the NaCl's containing 0g/L and 30g/L
There is obvious bacterium colony to grow on culture medium, there is micro bacterium colony to grow on the culture medium containing 50g/L, 70g/L and 100g/L NaCl；Through
After 5d culture, there is obvious bacterium colony to grow on the culture medium of the NaCl Han 0~70g/L, on the culture medium of the NaCl containing 100g/L
Micro bacterium colony growth can be observed.Confirm that bacterial strain F2 has certain salt tolerance, but too high in salinity will lead to its speed of growth and delay
Slowly.
Specific embodiment 4: present embodiment other style Shewanella Alishewanella sp.F2 denitrification capability
It is that 5% (v/v) is forwarded in sterilized different salinity denitrification culture medium with inoculum concentration, bottleneck is sealed, in 30
DEG C constant temperature stationary culture 5d, measures NO in each salinity culture solution2 --N、NO3 -- N concentration detects the denitrification capability of the bacterial strain.
After 5d is cultivated, bacterial strain F2 has certain NO under 0~70g/L NaCl Variation of Salinity Condition2 --N、NO3 -- N removal
Ability, but too high in salinity inhibits bacterial strain F2 growth, to reduce it to NO2 --N、NO3 -- N removal ability.In 0~30g/L
Under NaCl Variation of Salinity Condition, bacterial strain F2 has efficient NO2 --N、NO3 -- N removal ability, removal rate is 99% or so.30~
Under 100g/L NaCl Variation of Salinity Condition, too high in salinity inhibits bacterial strain F2 growth, causes bacterial strain F2 to NO2 --N、NO3 -- N removal ability
It significantly reduces.
Beneficial effects of the present invention are verified using following embodiment:
Embodiment one
Bacterial strain isolates and purifies:
(1) culture medium used in
A, bacterial strain denitrification culture medium (/L): by the CH of 5g3The K of COONa, 1g2HPO4, 0.8g NaNO2, 0.03g
CaCl2, 1g Na2CO3, 0.06g FeSO4·7H2O, the MgSO of 0.2g4·7H2O and 1000mL deionized water composition, pH=
10；
B, bacterial strain screening plating medium (/L): every liter of addition 20g fine jade on the basis of the bacterial strain denitrification culture medium
Rouge, pH=10；
C, bacterial strain preservation inclined tube culture medium (/L): identical as above-mentioned bacterial strains screening flat board culture medium.
Above-mentioned culture medium is before use, the 30min that sterilizes in the case where temperature is 121 DEG C of condition of high voltage.
(2) bacterial strain F2 is isolated and purified
Step 1: it draws 2mL and picks up from Zhuanghe City, Daliang City, Liaoning Province (39 ° 38 ' 31 of north latitude " 122 ° 58 ' 19 of east longitude ") sea wall
Muddy water sample be put into 250mL conical flask, the bacterial strain denitrification culture medium of 200mL is added, using sealed membrane by bottle sealing
Processing manufactures anaerobic environment；Enrichment culture liquid is obtained after constant temperature stationary culture 5~7 days under the conditions of temperature is 30 DEG C；
Step 2: gradient dilution is carried out using the enrichment culture liquid that coubling dilution obtains step 1, is drawn respectively each
Dilution suspension 0.5mL, will be rich with glass spreading rod into bacterial strain screening plating medium (having stood overnight, no varied bacteria growing)
After collecting culture solution coating uniformly, the screening flat board culture medium that the sterilized temperature of the second layer is not higher than 40 DEG C is poured into, manufactures anoxic
Environment is finally sealed around by plate using sealed membrane for cultivating denitrifying bacteria.It will be put down after the solidification of second layer culture medium
Plate is inverted, and being put in temperature is 30 DEG C of constant incubator, and culture is to growing obvious bacterium colony；
Step 3: the bacterial strain that picking step 2 is separated repeatedly is crossed pure on the double-deck bacterial strain screening plating medium
Change, until microscopically observation shows no miscellaneous bacteria, at this time it is believed that bacterial strain F2 of the invention has been purified and finished；Picking purifying
Afterwards come out bacterium colony, cross on bacterial strain preservation inclined tube culture medium, wait grow obvious bacterium colony be placed in 4 DEG C of refrigerators save it is standby
With.
The denitrification capability of bacterial strain measures
Enrichment culture is carried out to bacterial strain F2 of the present invention, seals bottleneck, in 30 DEG C of constant temperature stationary cultures, is occurred to culture medium muddy
It is turbid and generate after gas as enrichment seed bacterium solution；It is inoculated in 5% (v/v) inoculum concentration equipped with the sterilized denitrification of 200mL
Culture medium is fitted into 250mL conical flask, is placed in 30 DEG C of constant temperature sealing and standing culture 5d；Timing takes quantitative culture solution, measurement training
NO in nutrient solution2 -- N and NO3 -- N detects the denitrification capability of the bacterial strain.Not to be inoculated with the culture medium of denitrifying bacteria as blank
Control, every group parallel 3.
Bacterial strain F2 is measured to NO2 -The removal effect of-N is as shown in figure 4, whereinRepresent NO2 -- N concentration, ■ represent removal
Rate；Bacterial strain F2 is to NO3 -The removal effect of-N is as shown in figure 5, whereinRepresent NO3 -- N concentration, ■ represent removal rate；From figure
In as can be seen that NO after cultivating 36h, in denitrification culture medium3 -- N and NO3 -- N concentration is decreased obviously, and F2 pairs of bacterial strain
NO2 --N、NO3 -The removal rate of-N is respectively 74.31% and 74.56%；After cultivating 48h, bacterial strain F2 is to NO2 -The removal rate of-N
Reach maximum value, is 98.97% NO at this time3 -The removal rate of-N is 94.81%；After cultivating 96h, bacterial strain F2 is to NO3 -- N's
Removal rate reaches maximum value, is 99.05%, NO at this time2 -The removal rate of-N is 98.44%.
The Salt resistant test of bacterial strain
By changing the salinity of screening isolation medium, the growing state of bacterial strain is observed.
The screening isolation medium of different salinity gradient processed, i.e. NaCl concentration are respectively 0g/L, 30g/L, 50g/L, 70g/
L,100g/L；By on the culture medium flat plate of bacterial strain F2 streak inoculation to each salinity, it is placed in 30 DEG C of constant incubators and cultivates 5d, with
Whether bacterial strain grows the foundation as the whether resistance to corresponding salinity of the bacterial strain.Not to be inoculated with the culture medium of denitrifying bacteria as blank
Control, every group parallel 3.
The results are shown in Table 2, after 2d is cultivated, has obvious bacterium colony raw on the culture medium of the NaCl containing 0g/L and 30g/L
It is long, there is micro bacterium colony to grow on the culture medium containing 50g/L, 70g/L and 100g/L NaCl；After 5d is cultivated, containing 0g/L,
There is obvious bacterium colony to grow on the culture medium of 30g/L, 50g/L and 70g/L NaCl, it is considerable on the culture medium of the NaCl containing 100g/L
Micro bacterium colony growth is observed, illustrates that bacterial strain F2 of the present invention has certain salt tolerance, but too high in salinity will lead to its speed of growth
Slowly.
2 bacterial strain F2 of table growing state in different salinity gradient media
++: there is obvious bacterium colony in media surface, illustrates bacterial strain F2 normal growth；
+: there is micro bacterium colony in media surface, illustrates that bacterial strain F2 has growth phenomenon, but the speed of growth is slow；
: media surface does not occur bacterium colony, illustrates that bacterial strain F2 is not grown.
Influence of the salinity to the denitrification capability of bacterial strain of the present invention
The denitrification culture medium of different salinity gradient processed, i.e. NaCl concentration be respectively 0g/L, 30g/L, 50g/L, 70g/L and
100g/L.Bacterial strain F2 is forwarded in sterilized each salinity medium with 5% (v/v) inoculum concentration, bottleneck is sealed, in 30 DEG C of constant temperature
5~7d of stationary culture.Measure NO in culture solution2 --N、NO3 -- N concentration detects the denitrification capability of bacterial strain F2.It is anti-not to be inoculated with
The culture medium of nitrobacteria is as blank control, and every group parallel 3.
Bacterial strain F2 is measured to NO2 -The removal effect of-N is as shown in fig. 6, whereinRepresent NO2 -- N concentration, ■ represent removal
Rate；Bacterial strain F2 is to NO3 -The removal effect of-N is as shown in fig. 7, whereinRepresent NO3 -- N concentration, ■ represent removal rate, from figure
In as can be seen that bacterial strain F2 to NO2 --N、NO3 -The removal ability of-N is influenced significant obvious by salinity.After 5d is cultivated, contain 0g/L
In the denitrification culture solution of 30g/L NaCl, bacterial strain F2 is to NO2 --N、NO3 -The removal ability of-N is significant, NO2 --N、NO3 -- N is gone
Except rate is 99% or so, illustrate bacterial strain F2 normal growth, NO under the conditions of NaCl containing 30g/L2 --N、NO3 -- N removal ability
It is not influenced by salinity.In the culture solution of the NaCl containing 50g/L and 70g/L, bacterial strain F2 is to NO2 --N、NO3 -The removal ability of-N compared with
Difference, removal rate in 24~31% ranges, illustrate bacterial strain F2 slow growth, NO under the conditions of containing 50~70g/L NaCl2 --N、
NO3 -- N removal ability is influenced to be substantially reduced by salinity.In the culture solution of the NaCl containing 100g/L, bacterial strain F2 is to NO2 -- N removal
Rate is maintained at 26% or so, to NO3 -- N illustrates that bacterial strain F2 is grown under the conditions of NaCl containing 100g/L and is pressed down without removal ability
System, NO2 --N、NO3 -- N removal ability is seriously affected.
The best salt tolerant denitrogenation condition of bacterial strain
The sterilized denitrification of NaCl containing the 30g/L culture medium of 200mL is fitted into 250mL conical flask, 30 DEG C of sealing and standings
Bacterial strain F2 of the present invention is cultivated, investigates different pH (3.0,5.0,7.0,9.0,10.0,11.0) NaNO respectively2Initial concentration (0.4g/
L, 0.8g/L, 1.6g/L, 2.4g/L, 3.2g/L) and inoculum concentration (1% (v/v), 3% (v/v), 5% (v/v), 7% (v/v),
10% (v/v)) under the conditions of bacterial strain F2 to NO2 --N、NO3 -- N removal ability.
The result shows that bacterial strain F2 adapts to slight alkali environment under 30g/L NaCl Variation of Salinity Condition, it is 9~10 models in pH
In enclosing, bacterial strain F2 is to NO2 --N、NO3 -The removal rate of-N respectively reaches 99% and 96% or more；The optimum N aNO of bacterial strain F22Initially
Concentration is 0.4~0.8g/L, and bacterial strain F2 is to NO2 --N、NO3 -The removal rate of-N respectively reaches 98% and 94% or more；Bacterial strain F2's
Most suitable inoculum concentration is 5~7% (v/v), and bacterial strain F2 is to NO2 --N、NO3 -The removal rate of-N respectively reaches 99% and 96% or more.
Sequence table
<110>Northeast Institute of Geography and Agricultural Ecology, Chinese Academy of Sc
The denitrifying bacteria of<120>one plants of salt tolerants and its application
<160> 3
<210> 1
<211> 1033
<212> DNA
<213>other style Shewanella (Alishewanella sp.)
<220>
<223>other style Shewanella (Alishewanella sp.) F2
<400> 1
tagcggcgga cgggtgagta atgcgtagga agctgcccga tagaggggga taccagttgg 60
aaacgactgt taataccgca taatgtctac ggaccaaagt gtgggacctt cgggccacat 120
gctatcggat gcgcctacgt gggattagct agttggtggg gtaatggctc accaaggcga 180
cgatccctag ctggtttgag aggatgatca gccacactgg aactgagaca cggtccagac 240
tcctacggga ggcagcagtg gggaatattg gacaatgggc gcaagcctga tccagccatg 300
ccgcgtgtgt gaagaaggcc ttcgggttgt aaagcacttt cagtggggag gaagggtgtt 360
gtgttaatag tacagcactt tgacgttacc cacagaagaa gcaccggcta actccgtgcc 420
agcagccgcg gtaatacgga gggtgcaagc gttaatcgga attactgggc gtaaagcgca 480
cgcaggcggc tttttaagtc ggatgtgaaa gccccgggct caacctggga attgcatctg 540
atactgggaa gctagagtat gtgagagggg ggtagaattc caagtgtagc ggtgaaatgc 600
gtagagattt ggaggaatac cagtggcgaa ggcggccccc tggcacaata ctgacgctca 660
ggtgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg ccgtaaacga 720
tgtctactag ctgttcgcgg ccttgtgttg tgagtagcgc agctaacgca ttaagtagac 780
cgcctgggga gtacggtcgc aagattaaaa ctcaaatgaa ttgacggggg cccgcacaag 840
cggtggagca tgtggtttaa ttcgacgcaa cgcgaagaac cttacctact cttgacatct 900
acagaagaac gcagagatgt gtttgtgcct tcgggaactg taagacaggt gctgcatggc 960
tgtcgtcagc tcgtgttgtg aaatgttggg ttaagtcccg caacgagcgc aacccttatc 1020
cttagttgcc agc 1033
<210>2
<211> 20
<212> DNA
<213>artificial sequence
<220>
<223>forward primer 27F
<400>2
agagtttgatcctggctcag 20
<210>3
<211> 19
<212> DNA
<213>artificial sequence
<220>
<223>reverse primer 1492R
<400>3
ggttaccttgttacgactt 19
Claims (5)
1. the denitrifying bacteria of one plant of salt tolerant, it is characterised in that: the bacterial strain is other style Shewanella Alishewanella
Sp.F2, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation address is Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, the deposit date is on March 25th, 2019 is commonly micro- in China Committee for Culture Collection of Microorganisms
Bio-Centers preservation, deposit number are CGMCC No:17433.
2. the application of the denitrifying bacteria of salt tolerant described in claim 1 denitrogenation under the conditions of saliferous.
3. application according to claim 2, it is characterised in that the saliferous condition is NaCl concentration≤30g/L.
4. application according to claim 2, it is characterised in that the saliferous condition pH is 9~10, NaNO2Initial concentration is
0.4~0.8g/L.
5. application according to claim 2, it is characterised in that the inoculum concentration of the denitrifying bacteria of the salt tolerant is 5~7%
(v/v)。
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| CN111793573A (en) * | 2020-03-26 | 2020-10-20 | 大连理工大学 | Shewanella alga strain with functions of heterotrophic nitrate heterotrophic reduction and autotrophic nitrate heterotrophic reduction to ammonium, culture method and application thereof |
| CN112175934A (en) * | 2020-10-10 | 2021-01-05 | 中国科学院东北地理与农业生态研究所 | Microbial material with salt-tolerant denitrification capability and preparation method and application thereof |
| CN113151068A (en) * | 2021-04-01 | 2021-07-23 | 广东博沃特生物科技有限公司 | Shewanella denitrificans for degrading organic pollutants and application thereof |
| CN114686391A (en) * | 2020-12-31 | 2022-07-01 | 中国石油化工股份有限公司 | High-salinity-tolerance bacterium and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111793573A (en) * | 2020-03-26 | 2020-10-20 | 大连理工大学 | Shewanella alga strain with functions of heterotrophic nitrate heterotrophic reduction and autotrophic nitrate heterotrophic reduction to ammonium, culture method and application thereof |
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| CN112175934A (en) * | 2020-10-10 | 2021-01-05 | 中国科学院东北地理与农业生态研究所 | Microbial material with salt-tolerant denitrification capability and preparation method and application thereof |
| CN114686391A (en) * | 2020-12-31 | 2022-07-01 | 中国石油化工股份有限公司 | High-salinity-tolerance bacterium and application thereof |
| CN114686391B (en) * | 2020-12-31 | 2023-07-04 | 中国石油化工股份有限公司 | High-salt-tolerance bacterium and application thereof |
| CN113151068A (en) * | 2021-04-01 | 2021-07-23 | 广东博沃特生物科技有限公司 | Shewanella denitrificans for degrading organic pollutants and application thereof |
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