CN106047746B - One plant of pyridine degradable bites dyestuff bacterium and its application in wastewater treatment containing pyridine - Google Patents
One plant of pyridine degradable bites dyestuff bacterium and its application in wastewater treatment containing pyridine Download PDFInfo
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- CN106047746B CN106047746B CN201610319899.2A CN201610319899A CN106047746B CN 106047746 B CN106047746 B CN 106047746B CN 201610319899 A CN201610319899 A CN 201610319899A CN 106047746 B CN106047746 B CN 106047746B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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- C12N1/205—Bacterial isolates
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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
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- C02F2101/34—Organic compounds containing oxygen
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Abstract
The invention discloses one plant of pyridine degradables to bite dyestuff bacterium and its application in wastewater treatment containing pyridine.The mature aerobic particle mud that the present invention takes out from the sbr reactor device for removing pyridine directly screens, and it is separated by the screening and culturing medium of sole carbon source, nitrogen source of pyridine, pyridine degradable specific strain is obtained, it is to bite dyestuff bacterium Pigmentiphaga sp. through molecular biology identification, it is named as Pigmentiphaga sp.NJUST35, deposit number is CCTCC NO:M2016013.Pyridine degradable of the invention bites dyestuff bacterium, can be grown by sole carbon source and nitrogen source of pyridine.Pigmentiphaga sp.NJUST35 is added in pyridine industrial wastewater to be handled, pyridine degradable rate, COD removal rate and mineralized nitrogen rate are respectively 100%, 76.3% and 35.61%.The bacterial strain has the ability that well adapts to and tolerance performance with efficient pyridine degradable ability, high mineralization ability and to the toxicity of pyridine, has a good application prospect in the processing of high-purity pyridine waste water.
Description
Technical field
The invention belongs to environmental organic pollutant biologic treating technique fields, and in particular to one plant of pyridine degradable bites dyestuff bacterium
And its application in Industrial Wastewater Treatment containing pyridine.
Background technique
Pyridine can produce synthesis multiple compounds, including Insecticides (tech) & Herbicides (tech), medicine as a kind of important industrial chemicals
Product, disinfectant, dyestuff etc. are widely used in the industries such as medicine, pesticide, chemical industry.Pyridine is water-soluble strong, easily in soil and groundwater
Between shift；Stable structure has bio-toxicity, teratogenesis and carcinogenic nature.Therefore, it repairs and is polluted by pyridine heterocyclic compounds
Soil and water environment, it has also become one of the important topic of field of environmental improvement at present.
Currently, the physical chemistry processing technique such as processing technique of pyridine waste water predominantly absorption, burning and oxidation.However,
Materializing strategy technical costs is generally higher, is only applicable to the processing of high-purity pyridine waste water, and pyridine in treated waste water
Still possible abundant residues, it is difficult to realize qualified discharge.Compared to physico-chemical process, biological degradation method have it is economical, efficiently and
The advantages that without secondary pollution is most widely used wastewater processing technology, it can be achieved that full innoxious governance.But due to pyridine
Bio-toxicity, the biochemical treatment of pyridine have very big technical difficulty, need under given conditions, can using specified microorganisms
It realizes.The pyridine degradable special efficacy Pseudomonas reported includes Arthrobacter, Rhodococcus opacus at present,
Pseudomonas, Shinella, Bacillus, Paracoccus, Streptomyces, Shewanella, Nocardiodes
Deng, but most of pyridine degradable bacteria is low to the tolerable concentration of pyridine (being less than 1000mg/L), and degradation property is restricted, it is difficult to full
The requirement of sufficient practical engineering application.
Therefore, screening degradation efficiency is high, tolerable concentration is high, adapts to the pyridine degradable bacteria of true environment, for solving pyridine
Waste water pollution problem has a very important significance.
Summary of the invention
The purpose of the present invention is to provide one plant of pyridine degradables to bite dyestuff bacterium Pigmentiphaga sp.NJUST35, can
Overcome existing pyridine degradable bacteria strain limited amount, and low to pyridine tolerable concentration, treatment effeciency is low, and bacterial strain type is more single
One deficiency.
The maturation that inventor takes out from the sbr reactor device for removing pyridine for having run 2 years in laboratory is aerobic
Granule sludge directly screens, and is separated by the screening and culturing medium of sole carbon source, nitrogen source of pyridine, and pyridine degradable spy has been obtained
Bacterial strain NJUST35 is imitated, is to bite dyestuff bacterium Pigmentiphaga sp. through molecular biology identification, is named as Pigmentiphaga
Sp.NJUST35, GenBank accession number are KU051387.The bacterial strain is on January 6th, 2016 in China typical culture collection
Center (CCTCC) preservation, deposit number are CCTCC NO:M 2016013.
The present invention also provides the above-mentioned dyestuff bacterium Pigmentiphaga sp.NJUST35 that bites in wastewater treatment containing pyridine
Using what strain can be used for pyridine wastewater treatment headed by the bacterial strain bites dyestuff bacterium Pigmentiphaga sp..
Above-mentioned pyridine degradable bites application of the dyestuff bacterium Pigmentiphaga sp.NJUST35 in wastewater treatment containing pyridine,
Method particularly includes: pyridine degradable is bitten into dyestuff bacterium Pigmentiphaga sp.NJUST35 seed liquor and is inoculated into waste water containing pyridine
Culture, the temperature of culture are 25 DEG C~35 DEG C, and the pH of the waste water containing pyridine is neutrality.
Above-mentioned pyridine degradable bites dyestuff bacterium Pigmentiphaga sp.NJUST35 answering in wastewater treatment containing pyridine
In, the temperature of culture is preferably 30 DEG C.
Pigmentiphaga sp.NJUST35 of the invention, can be grown by sole carbon source and nitrogen source of pyridine.
Pigmentiphaga sp.NJUST35 is added in pyridine industrial wastewater to be handled, pyridine degradable rate, COD removal rate and ammonia
Conversion rate of nitrogen is respectively 100%, 76.3% and 35.61%.The bacterial strain have efficient pyridine degradable ability, high mineralization ability with
And there is the ability that well adapts to and tolerance performance to the toxicity of pyridine, have in the processing of high-purity pyridine waste water good
Application prospect.
Detailed description of the invention
Fig. 1 is the transmission electron microscope picture of bacterial strain Pigmentiphaga sp.NJUST35 of the present invention.
Fig. 2 is bacterial strain Pigmentiphaga sp.NJUST35 of the present invention in the MSM that pyridine initial concentration is 1000mg/L
In to the release conditions result figure of ammonia nitrogen in the degradation and degradation process of pyridine.
Specific embodiment
The following examples can make those skilled in the art that the present invention be more fully understood, but not limit in any way
The present invention.
Embodiment 1
The screening of Pigmentiphaga sp.NJUST35 separates and identification.
(1) screening and separation of bacterial strain
Mature aerobic particle mud is taken out from the existing sbr reactor device for handling pyridine waste water, it is abundant with mortar
It is diluted to 10 respectively using gradient dilution method with sterile water after grinding4-1010Times.LB solid medium is prepared, after sterilizing while hot
It pours into the culture dish of same sterilized 90 × 90cm, 20 μ L previously diluted sample is respectively coated after solidification to be cooled, is placed in
3d is cultivated under conditions of 30 DEG C in biochemical cultivation case.The bacterial strain that colony characteristics on culture dish have notable difference is selected, is used
Plate streaking separation method purified, continuous purification three times after, obtain single bacterial strain, and carry out inclined-plane preservation.Picking point
From obtained single colonie, it is respectively connected to be to be put into full Wen Zhen in sole carbon source and the minimal medium MSM of nitrogen source containing pyridine
It swings in incubator at 30 DEG C, is cultivated under the conditions of 180r/min, monitor the concentration value of pyridine.It chooses so that pyridine is sent out in culture medium
The bacterial strain of raw obvious degradation, is named as NJUST35, is carried out inclined-plane and saves the cryo-conservation with -80 DEG C to further grind
Study carefully.
The composition of minimal medium MSM is as follows: Na2HPO4·12H2O (1.529g/L), KH2PO4(0.372g/L),
MgSO4·7H2O (0.1g/L), CaCl2(0.05g/L), trace element solution SL-4 (10mL/L).Microelement SL-4 composition:
EDTA(0.5g/L),FeSO4·7H2O (0.2g/L), microelement SL-6 (100mL/L).Microelement SL-6 composition:
ZnSO4·7H2O (0.01g/L), MnCl2·4H2O (0.03g/L), H3BO4(0.3g/L), CoCl2·6H2O (0.2g/L),
CuCl2·2H2O(0.01g/L),NiCl2·6H2O (0.02g/L), Na2MoO4·2H2O(0.03g/L)。
(2) identification of bacterial strain
Morphology, Physiology and biochemistry test are carried out to bacterium.The 16S rDNA sequence for measuring bacterial strain, by the 16S rDNA of bacterial strain
Sequence in gene order and world GenBank database carries out online tetraploid rice, and finally determining from molecular level should
The kind of bacterium.
1. morphological feature: NJUST35 bacterium colony surface glossy clear, neat in edge are mixed in diffusivity in liquid medium
It is turbid.The strain cell is in the shape of a rod, and surface is smooth.Having a size of 1.11-1.20 μ m 561-616nm.Fig. 1 is the transmission electron microscope of bacterium
Photo.
2. physiological and biochemical property: Gram-negative, urease negative, amino acid decarboxylases is negative, arginine dihydrolase yin
Property, enzyme positive is contacted, oxidase positive can utilize citric acid, can utilize malic acid, can utilize adipic acid, aesculin can be hydrolyzed,
It is unable to gelatin liquefaction, nitrate cannot be restored, be unable to hydrolyzed casein, starch cannot be hydrolyzed, tyrosine cannot be hydrolyzed, it is aerobic.
3. molecular biology identification: using the core DNA of NJUST35 bacterium as template, with the logical of the PCR amplification of 16S rRNA gene
It is primer with primer, carries out PCR amplification, measure its complete sequence, the 16S rDNA gene order of the bacterial strain is shown in sequence table.By bacterial strain
16S rDNA gene order be committed to GenBank database (the GenBank number of logging in be KU051387), with GenBank data
Sequence in library carries out online tetraploid rice, the results showed that, NJUST35 and Pigmentiphaga sp.Zn-d-2 and
The sequence similarity of Pigmentiphaga sp.NDS-1 is up to 99% or more.
It is accredited as according to the morphology of NJUST35, Physiology and biochemistry test and molecular biological analysis, NJUST35
Pigmentiphaga sp. is named as Pigmentiphaga sp.NJUST35.
Embodiment 2
Pigmentiphaga sp.NJUST35 bacterial strain studies the degradation property of pyridine.
Pyridine degradable bacterial strain NJUST35 is seeded to the LB culture medium of addition 1000mg/L pyridine, under the conditions of 30 DEG C with
The revolving speed shaking table culture of 180r/min, carries out the enrichment culture of NJUST35, enters logarithmic growth phase later period (about 3d) to thallus,
The gained thallus revolving speed of 8000rpm is centrifuged 15 minutes, supernatant is skimmed, is hanged thallus again using the method for the concussion that is vortexed
Float on sterile liquid MSM, is centrifuged.Repeated washing process three times after, by thallus be resuspended in sterile liquid MSM (adjust be added
MSM amount, control bacterium suspension OD600About 1.0), seed liquor is obtained.The LB culture medium that the present invention uses is the art
Conventional use of culture medium.
The liquid MSM for containing 1000mg/L pyridine is prepared as simulated wastewater, simulation pyridine is added in above-mentioned seed liquor and is given up
In water, inoculum concentration 5% observes in degradation process pyridine in waste water with the revolving speed shaking table culture of 180r/min under the conditions of 30 DEG C
TOC concentration when concentration, ammonia nitrogen concentration and degradation terminal.Set up the blank control for not being inoculated with NJUST35.Experimental result is as schemed
Shown in 2.As shown in Figure 2,1000mg/L pyridine can be in degradable in 148h；Nitrogen during pyridine degradable, in pyridine structure
It is released in the form of ammonia nitrogen, the conversion ratio of ammonia nitrogen is about 36.68% after pyridine degradable；TOC is significantly reduced, removal rate
Up to 85.70%；And in the control sample for not being inoculated with NJUST35, pyridine does not obtain obvious degradation.
This example demonstrates that it is sole carbon source that isolated Pigmentiphaga sp.NJUST35, which can use pyridine,
Growth and breeding is carried out with nitrogen source, and can realize the mineralising of pyridine.
Embodiment 3
Processing of the Pigmentiphaga sp.NJUST35 to the industrial wastewater containing pyridine.
Pigmentiphaga sp.NJUST35 seed liquor is accessed into the actual industrial waste water containing pyridine with 5% inoculum concentration
(989mg/L containing pyridine, COD1328mg/L, total phosphorus 55mg/L), with the revolving speed shaking table culture of 180r/min under the conditions of 30 DEG C.
Monitor wastewater treatment front and back COD value, pyridine and ammonia nitrogen concentration.
As shown in Table 1, industrial wastewater containing pyridine is realized after bacterial strain Pigmentiphaga sp.NJUST35 handles 108h
Pyridine it is degradable, COD value is reduced to 314.67mg/L, and ammonia nitrogen burst size is 63.11mg/L.Pyridine degradable rate, COD removal
Rate and mineralized nitrogen rate are respectively 100%, 76.3% and 35.61%.
This example demonstrates that isolated Pigmentiphaga sp.NJUST35 can be successfully applied to containing pyridine industry
The efficient removal of pyridine and COD in industrial wastewater containing pyridine is realized in the biochemical treatment of waste water.
The situation of change of indices before and after 1 Industrial Wastewater Treatment containing pyridine of table
SEQUENCE LISTING
<110>Institutes Of Technology Of Nanjing
<120>one plants of pyridine degradables bite dyestuff bacterium and its application in wastewater treatment containing pyridine
<130> 2016
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1397
<212> DNA
<213> Pigmentiphaga
<400> 1
tgcaagtcga acggcagcgc ggacttcggt ctggcggcga gtggcgaacg ggtgagtaat 60
gcatcggaac gtgcccagta gcgggggata actacgcgaa agcgtagcta ataccgcata 120
cgccctacgg gggaaagggg gggatcgcaa gacctctcac tattggagcg gccgatgtcg 180
gattagctag ttggtggggt aaaggcctac caaggcgacg atccgtagct ggtttgagag 240
gacgaccagc cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtggg 300
gagattttgg acaatggggg caacctgatc cagccatgcc gcgtgtgcga agaaggcttc 360
gggtgtaaag cactttcggc aggaaagaaa cggcgccggg ctaatatctg gcgtaactga 420
cggtacctgc agaataagca ctggctaact acgtgccagc agccgcggta atacgtaggg 480
tgcaagcgtt aatcggaatt actgggcgta aagcgtgcgc aggcggttcg gaaagaaaga 540
tgtgaaatcc cagggctcaa ccttggaact gcatttttaa ctcccgaact agagtgtgtc 600
agaggggggg tggaattcca cgtgtagcag tgaaatgcgt agatatgtgg aggaacaccg 660
atggcgaagg cagccccctg ggataacact gacgctcatg cacgaaagcg tggggagcaa 720
acaggattag ataccctggt agtccacgcc ctaaacgatg tcaactagct gttgggttct 780
tcggagcttg gtagcgcagc taacgcgtga agttgaccgc ctggggagta cggtcgcaag 840
attaaaactc aaaggaattg acggggaccc gcacaagcgg tggatgatgt ggattaattc 900
gatgcaacgc gaaaaacctt acctaccctt gacatgtctg gaatcccgaa gagatttggg 960
agtgctcgaa agagaaccgg aacacaggtg ctgcatggcc gtcgtcagct cgtgtcgtga 1020
gatgttgggt taagtcccgc aacgagcgca acccttgtca ttagttgcta cgaaagggca 1080
ctctaatgag actgccggtg acaaaccgga ggaaggtggg gatgacgtca ggtcctcatg 1140
gcccttatgg gtagggcttc acacgtcata caatggtcgg gacagagggc agccaacccg 1200
cgagggggag ccaatcccag aaacccgatc gtagtccgga ttgcagtctg caactcgact 1260
gcatgaagtc ggaatcgcta gtaatcgcgg atcagcatgt cgcggtgaat acgttcccgg 1320
gtcttgtaca caccgcccgt cacaccatgg gagtgggttc taccagaagt aggtagccta 1380
accgcaagga gggcgct 1397
Claims (4)
1. one plant of pyridine degradable bites dyestuff bacterium, to bite dyestuff bacterium Pigmentiphaga sp.NJUST35, deposit number CCTCC
NO:M 2016013.
2. pyridine degradable as described in claim 1 bites application of the dyestuff bacterium in wastewater treatment containing pyridine.
3. pyridine degradable as claimed in claim 2 bites application of the dyestuff bacterium in wastewater treatment containing pyridine, which is characterized in that tool
Body method are as follows: pyridine degradable is bitten into dyestuff bacterium Pigmentiphaga sp.NJUST35 seed liquor be inoculated into waste water containing pyridine and train
It supports, the temperature of culture is 25 DEG C~35 DEG C, and the pH of the waste water containing pyridine is neutrality.
4. pyridine degradable as claimed in claim 3 bites application of the dyestuff bacterium in wastewater treatment containing pyridine, which is characterized in that institute
The cultivation temperature stated is 30 DEG C.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103540544A (en) * | 2013-08-23 | 2014-01-29 | 南京理工大学 | Bacillus radicicola capable of degrading pyridine as well as breeding method and application thereof |
| CN105802894A (en) * | 2016-05-13 | 2016-07-27 | 南京理工大学 | Pyridine degrading bacteria and application thereof in pyridine-containing wastewater treatment |
| CN107935309A (en) * | 2017-12-01 | 2018-04-20 | 南京理工大学 | Priming produces waste water physicochemical pre-treat biological intensive treatment integrated technique |
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- 2016-05-13 CN CN201610319899.2A patent/CN106047746B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103540544A (en) * | 2013-08-23 | 2014-01-29 | 南京理工大学 | Bacillus radicicola capable of degrading pyridine as well as breeding method and application thereof |
| CN105802894A (en) * | 2016-05-13 | 2016-07-27 | 南京理工大学 | Pyridine degrading bacteria and application thereof in pyridine-containing wastewater treatment |
| CN107935309A (en) * | 2017-12-01 | 2018-04-20 | 南京理工大学 | Priming produces waste water physicochemical pre-treat biological intensive treatment integrated technique |
Non-Patent Citations (2)
| Title |
|---|
| Genbank.Pigmentiphaga sp.NJUST35 16S ribosomal RNA gene,partial sequence * |
| Genbank:KU051387.1.《NCBI》.2016, * |
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