CN106119159A - The comamonas of one high-efficiency degradation pyridine carboxylic acid and application thereof - Google Patents
The comamonas of one high-efficiency degradation pyridine carboxylic acid and application thereof Download PDFInfo
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- CN106119159A CN106119159A CN201610497030.7A CN201610497030A CN106119159A CN 106119159 A CN106119159 A CN 106119159A CN 201610497030 A CN201610497030 A CN 201610497030A CN 106119159 A CN106119159 A CN 106119159A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
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Abstract
The invention provides comamonas and the application thereof of a high-efficiency degradation pyridine carboxylic acid, belong to technical field of bioengineering.The present invention separates a strain pyridine carboxylic acid degradation bacteria, comamonas, and its Classification And Nomenclature is Comamonas sp., is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC NO.10700.It is an advantage of the invention that this comamonas can be at the pH value of relative broad range, pyridine carboxylic acid of effectively degrading in the case of particularly basicity is higher；This comamonas can be administered as the aerobe of biological reinforced formulation application pyridine carboxylic acid waste water in weak base to weakly acidic condition, has broad application prospects.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a high-efficiency degradation pyridine carboxylic acid comamonas and
Its application.
Background technology
Pyridine carboxylic acid and derivant thereof are widely used in the production of medicine, pesticide and household chemicals, pyridine carboxylic acid
Extensively application causes it to pollute environment.Pyridine carboxylic acid waste water can be processed by aerobe method.At present, aerobic degradation pyridine
The microbial resources of formic acid mainly have arthrobacterium, mycobacteria and streptomycete etc..The bacterial strain report planted is belonged to the most few about other
See.Additionally, the pyridine carboxylic acid degradation bacteria that document is reported all grows, substantially 6.0 to 7.0 in the range of comparatively gentle pH
In the range of, and actual pyridine carboxylic acid industrial wastewater pH excursion is wider, the pyridine carboxylic acid degradation bacteria that therefore document is reported is real
Border application is restricted.
Summary of the invention
It is an object of the invention to provide comamonas and the application thereof of a high-efficiency degradation pyridine carboxylic acid, it is possible to relatively
Pyridine carboxylic acid is efficiently removed in wide ph range.
For reaching above-mentioned purpose, the technical solution used in the present invention is:
The comamonas of one high-efficiency degradation pyridine carboxylic acid, its Classification And Nomenclature is Comamonas sp., is preserved in China
Microbiological Culture Collection administration committee common micro-organisms center, preserving number is CGMCC NO.10700.
The application in terms of degraded pyridine carboxylic acid of the comamonas of described efficient degradation pyridine carboxylic acid.
The comamonas of described efficient degradation pyridine carboxylic acid is in terms of preparing biological bacteria preparation or biological reinforced preparation
Application.
The comamonas of described efficient degradation pyridine carboxylic acid as biological bacteria preparation or biological reinforced preparation at pyridine
Application in formic acid wastewater process.
Pyridine carboxylic acid be sole carbon source, nitrogen source and the energy, pH be 7, under conditions of temperature is 30 DEG C, comamonas energy
The enough waste water to pyridine carboxylic acid concentration≤800mg/L is degraded.
Pyridine carboxylic acid be sole carbon source, nitrogen source and the energy, temperature be 30 DEG C, under conditions of pH is 5～9, Comamonas
The waste water of pyridine carboxylic acid concentration≤200mg/L can be degraded by bacterium.
Relative to prior art, the invention have the benefit that
The present invention is directed to the problem that the pyridine carboxylic acid degradation bacteria of previous literature report exists, tamed by scientific method, sieve
Selecting the pyridine carboxylic acid degradation bacteria comamonas that a strain is new, its Classification And Nomenclature is Comamonas sp., is preserved in China micro-
Biological inoculum preservation administration committee's common micro-organisms center, preserving number is CGMCC NO.10700.This comamonas can be
Pyridine carboxylic acid is efficiently removed in the range of wider pH value, thus degradation treatment pyridine first be applicable to actual pyridine carboxylic acid industrial wastewater
Acid.The comamonas that the present invention provides can be added in existing pyridine carboxylic acid Waste Water Treatment as biological bacteria preparation,
Strengthen the disposal ability of former processing system；The comamonas that the present invention provides can be as biological reinforced formulation application in weak base
To weakly acidic condition, the aerobe of pyridine carboxylic acid waste water is administered, and has wide application to dive in the process of pyridine carboxylic acid waste water
Power.
Further, being found through experiments, the comamonas that the present invention provides is at pH=7, temperature 30 DEG C, pyridine carboxylic acid
Under conditions of initial concentration≤800mg/L, it is possible to degradable pyridine carboxylic acid；It addition, be 5～9 at pH, pyridine carboxylic acid is unique
Carbon, nitrogen, the energy, under conditions of temperature 30 DEG C, and pyridine carboxylic acid initial concentration≤200mg/L, this comamonas can be complete
Degraded pyridine carboxylic acid.
Preservation explanation
The comamonas of efficient degradation pyridine carboxylic acid of the present invention has carried out following preservation:
The preservation time: on April 9th, 2015, preservation place: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese micro-life
Thing culture presevation administration committee's common micro-organisms center, CGMCC；Preserving number is CGMCC NO.10700.
Accompanying drawing explanation
Fig. 1 be the comamonas that provides of the present invention with pyridine carboxylic acid as sole carbon, nitrogen, the energy and pyridine carboxylic acid initial
Concentration is the colony morphology characteristic in the Solid agar culture of 100mg/L: after cultivating 3 days at 30 DEG C, bacterium colony is rice white, limit
Edge is irregular, rough surface is rough, surface is not swelled but combines closely in media surface, and difficult when inoculation
To provoke.
Fig. 2 be the comamonas that provides of the present invention with pyridine carboxylic acid as sole carbon, nitrogen, the energy (initial concentration
Fluid medium 100mg/L), cultivating the scanning electron microscope (SEM) photograph after 10h at 30 DEG C, cell is rod-short, and length dimension is 1～2 μ
m。
Fig. 3 be the comamonas that provides of the present invention with pyridine carboxylic acid as sole carbon, nitrogen, the energy, pH is 7, temperature 30
DEG C, pyridine carboxylic acid mass concentration be 800mg/L fluid medium in growth with pyridine carboxylic acid degradation curve figure.
Fig. 4 be the comamonas that provides of the present invention at various ph values, temperature 30 DEG C, pyridine carboxylic acid be sole carbon, nitrogen,
When the energy and pyridine carboxylic acid initial concentration are 200mg/L, the degradation curve figure of pyridine carboxylic acid.
Detailed description of the invention
Below in conjunction with technical scheme and accompanying drawing, the present invention is further elaborated.
1, the comamonas that the present invention provides, its screening step is as follows:
From the Aeration tank of Xi'an Medicine Factory, it is collected oxygen activity mud sample, carries out taming, cultivating as bacterium source.Will
50mL activated sludge, the sodium pyrophosphate of addition 0.2% is as solving flocculant, and activity dirt is smashed in vibration 5～10min on the oscillator
After mud flco, take 5mL activated sludge, add in the shaking flask containing 95mL minimal medium, add pyridine carboxylic acid as uniquely
Carbon, nitrogen, the energy, wherein the concentration of pyridine carboxylic acid is 100mg/L, carries out shaking table cultivation, temperature 30 DEG C, rotating speed 180r/min.Nothing
Machine salt culture medium composition: Na2HPO4·12H2O, 7g/L；KH2PO4, 1g/L；CaCl2·2H2O, 10mg/L；FeCl3, 2mg/L；
MgSO4·7H2O, 20mg/L.When bacterium solution becomes muddy, and pyridine carboxylic acid can't detect, and transfers next time；Tame and docile through constantly
Change and cultivate, finally give with pyridine carboxylic acid as sole carbon, the stable bacterium solution of nitrogen, the energy.To stablize bacterium solution dilution, with pyridine first
Acid is to be coated on the solid medium of sole carbon, nitrogen, the energy.Solid culture based component: Na2HPO4·12H2O, 7g/L；
KH2PO4, 1g/L；CaCl2·2H2O, 10mg/L；FeCl3, 2mg/L；MgSO4·7H2O, 20mg/L；Pyridine carboxylic acid, 100mg/L；
Agar 2%.This solid medium is put in illumination box, 30 DEG C, cultivate 4～5 days, choose single bacterium colony；Separate one
Strain with pyridine carboxylic acid as sole carbon, the comamonas of nitrogen, the energy；This bacterial strain is identified by 16S rDNA sequence, in
It is preserved in that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", its preserving number is on April 9th, 2015
CGMCC No.10700, Classification And Nomenclature is Comamonas sp..Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postal
Compile: 100101.
Growth pH wider range of this comamonas, all has preferably growth in the range of the pH of 5.0～9.0, the suitableeest
PH value is 7.0；Its morphological characteristic: with pyridine carboxylic acid as sole carbon, the fluid medium of nitrogen, the energy, at 30 DEG C cultivate 10h
After, as in figure 2 it is shown, cell is rod-short, length dimension is 1～2 μm；With pyridine carboxylic acid as sole carbon, the agar of nitrogen, the energy
On solid medium, cultivate after 3 days at 30 DEG C, as it is shown in figure 1, bacterium colony to be rice white, edge irregular, rough surface not only
Sliding, surface is not swelled but is combined closely in media surface, and is difficult to provoke when inoculation.This comamonas was both
Can grow in eutrophy culture medium LB, it is possible to be the inorganic of sole carbon source at small organic molecules such as glucose, sucrose, starch
Salt culture medium grows.
The 16S rDNA amplification of the comamonas that the present invention provides: by expanding the 16S rDNA of this bacterial strain, obtain
The 16S rDNA sequence of a length of 1428bp.PCR primer uses universal primer 27F (the 5 '-AGT TTG ATC of 16S rDNA
MTG GCT CAG-3 ') and 1492R (5 '-GGT TAC CTT GTT ACG ACT T-3 ').With PCR instrument (Applied
Biosystems, 2720thermal cycler) carry out amplified reaction.Table 1 and 2 respectively illustrates PCR reaction system and anti-
Answer condition.
Table 1PCR reaction system
Table 2 reaction condition
Take PCR primer 1 μ L and carry out 1% agarose gel electrophoresis, reclaim test kit (SK8131 glue reclaims test kit) with DNA
Reclaim purpose fragment, PCR primer is cloned on pGEM-T carrier, convert competent escherichia coli cell.Genbank enters
Row BLAST comparison.With MEGA 5.1 software building phylogenetic tree, use Neighbo-Joining method to analyze bacterial strain relationship and close
System (Bootstrap value=1000).Serial No. KP900021 logged in Genbank, with Comamonas
Sp.JC13 homology reaches 99%, infers that this bacterial strain belongs to Comamonas (Comamonas).
2, the cellular liquid culture of comamonas is prepared:
Picking comamonas list bacterium colony on the solid medium that pyridine carboxylic acid is sole carbon, nitrogen, the energy, loading is gone out
Bacterium with pyridine carboxylic acid as sole carbon, in the fluid medium of nitrogen, the energy, wherein the concentration of pyridine carboxylic acid is 100mg/L, in 30
DEG C, pH=7.0,180r/min, carry out aerobic cultivation 12～30 hours, take cultured bacterium solution 1mL and be seeded in 100mL containing pyridine
In the minimal medium of formic acid (200mg/L), 30 DEG C, pH=7.0,180r/min, carry out aerobic cultivation 20 hours, obtain clump
The cellular liquid culture of hair Zymomonas mobilis.
3, about the comamonas research to pyridine carboxylic acid aerobic degradation, its step is as follows:
[1] in 250mL conical flask, add 90mL minimal medium, add the pyridine carboxylic acid of 800mg/L as only
One carbon, nitrogen, the energy, sterilizing at 121 DEG C；
[2] the cellular liquid culture of the 10mL comamonas prepared is added in above-mentioned 90mL culture medium, 30 DEG C,
PH=7,180r/min, aerobic cultivation, result is as shown in Figure 3.
Fig. 3 be the comamonas that provides of the present invention with pyridine carboxylic acid as sole carbon, nitrogen, the energy, pH is 7, temperature 30
DEG C, pyridine carboxylic acid mass concentration be 800mg/L fluid medium in growth with pyridine carboxylic acid degradation curve figure, indicate feathering
The growth of Zymomonas mobilis and pyridine carboxylic acid are degraded situation, abscissa h express time in Fig. 3, and unit is hour, and left side vertical coordinate represents
The concentration of pyridine carboxylic acid, unit is mg/L, and right side vertical coordinate represents dry cell weight, and unit is mg/L；■ represents pyridine first
Acid concentration, represents thalli growth amount.From figure 3, it can be seen that comamonas pH be 7, temperature 30 DEG C, pyridine first
Under conditions of acid mass concentration≤800mg/L can degradable pyridine carboxylic acid, therefore, it is possible to as biological reinforced preparation to pyrrole
The pyridine carboxylic acid waste water of pyridine formic acid concn≤800mg/L processes.
4, the research to pyridine carboxylic acid aerobic degradation at various ph values about comamonas, its step is as follows:
[1] secure ph is the minimal medium of 3,4,5,6,7,9,10,12 respectively, adds the pyridine first of 200mg/L
Acid is as sole carbon, nitrogen, the energy, sterilizing at 121 DEG C；
[2] the cellular liquid culture of the 10mL comamonas prepared is separately added into the 90mL of above-mentioned different pH value
In culture medium, 30 DEG C, 180r/min, aerobic cultivation, result is as shown in Figure 4.
Fig. 4 be the comamonas that provides of the present invention at various ph values, temperature 30 DEG C, pyridine carboxylic acid be sole carbon, nitrogen,
When the energy and pyridine carboxylic acid initial concentration are 200mg/L, the degradation curve figure of pyridine carboxylic acid, indicate comamonas in difference
Growth under pH value and pyridine carboxylic acid are degraded situation, abscissa h express time in Fig. 4, and unit is hour, and vertical coordinate represents pyridine
The degradation rate of formic acid, unit is %.As can be seen from Figure 4: comamonas is 5～9 at pH value, temperature 30 DEG C, pyridine first
Under conditions of acid concentration≤200mg/L can degradable pyridine carboxylic acid, therefore, it is possible to as biological reinforced preparation to wider model
The pyridine carboxylic acid waste water that the pH value enclosed, particularly basicity are higher carries out degradation treatment.
The above, be only presently preferred embodiments of the present invention, not impose any restrictions the present invention, every according to the present invention
Any simple modification, change and the equivalent structure transformation that above example is made by technical spirit, all still falls within skill of the present invention
In the protection domain of art scheme.
Claims (6)
1. the comamonas of a high-efficiency degradation pyridine carboxylic acid, it is characterised in that: its Classification And Nomenclature is Comamonas sp.,
Being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC NO.10700.
2. the comamonas of the efficient degradation pyridine carboxylic acid described in claim 1 application in terms of degraded pyridine carboxylic acid.
3. the comamonas of the efficient degradation pyridine carboxylic acid described in claim 1 is preparing biological bacteria preparation or biological reinforced system
Application in terms of agent.
4. the comamonas of the efficient degradation pyridine carboxylic acid described in claim 1 is as biological bacteria preparation or biological reinforced preparation
Application in pyridine carboxylic acid waste water processes.
Apply the most as claimed in claim 4, it is characterised in that: pyridine carboxylic acid be sole carbon source, nitrogen source and the energy, pH be 7,
Under conditions of temperature is 30 DEG C, the waste water of pyridine carboxylic acid concentration≤800mg/L can be degraded by comamonas.
Apply the most as claimed in claim 4, it is characterised in that: it is that sole carbon source, nitrogen source and the energy, temperature are at pyridine carboxylic acid
30 DEG C, under conditions of pH is 5～9, the waste water of pyridine carboxylic acid concentration≤200mg/L can be degraded by comamonas.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112680384A (en) * | 2021-01-29 | 2021-04-20 | 微米环创生物科技(北京)有限公司 | Phenol degrading bacterium and application thereof |
| CN116354520A (en) * | 2021-05-07 | 2023-06-30 | 茅台学院 | Application of monascus comosus in treatment of ultra-high concentration white spirit wastewater |
| CN116354520B (en) * | 2021-05-07 | 2024-10-29 | 茅台学院 | Application of monascus comosus in treatment of ultra-high concentration white spirit wastewater |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112680384A (en) * | 2021-01-29 | 2021-04-20 | 微米环创生物科技(北京)有限公司 | Phenol degrading bacterium and application thereof |
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| CN116354520A (en) * | 2021-05-07 | 2023-06-30 | 茅台学院 | Application of monascus comosus in treatment of ultra-high concentration white spirit wastewater |
| CN116354520B (en) * | 2021-05-07 | 2024-10-29 | 茅台学院 | Application of monascus comosus in treatment of ultra-high concentration white spirit wastewater |
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