CN114456974B - Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment - Google Patents
Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment Download PDFInfo
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Classifications
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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
- 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
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/34—Organic compounds containing oxygen
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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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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Abstract
The invention relates to the technical field of DMF degradation, and provides a Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment. The Paracoccus thiooxidans is named 175A1-1, and has been preserved in China general microbiological culture Collection center (CGMCC) No.23659 at 10 and 25 of 2021, and the microorganism classification is named Paracoccus thiooxidansParacoccus sulfuroxidans. The Paracoccus thiooxidans 175A1-1 has higher DMF tolerance, and can convert DMF into NH by taking DMF as the sole carbon source and nitrogen source 3 -N, achieving efficient ammoniation and mineralization of DMF; moreover, the strain can be applied to site engineering efficiently and stably.
Description
Technical Field
The invention relates to the technical field of DMF degradation, in particular to a Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment.
Background
N, N-Dimethylformamide (DMF) is an organic solvent with stable chemical property, high boiling point and excellent performance. Besides halogenated hydrocarbon, the water-soluble organic solvent can be mutually dissolved with water and a plurality of organic solvents in any proportion, and the water-soluble organic solvent is called as a universal solvent due to the good dissolving capacity of the water-soluble organic solvent, and is also widely applied to industries such as synthetic textile, leather, electronics, pharmacy, pesticide and the like. DMF is therefore also one of the common high concentration contaminants in industrial wastewater.
The DMF wastewater treatment method includes physical method (such as adsorption method and membrane method), chemical method (such as photocatalytic oxidation, plasma oxidation, catalytic wet oxidation and chemical extraction), and biological method. Biological processes are considered to be a better treatment method due to economy and environmental protection. However, due to poor biochemical property of DMF and biological toxicity of degradation products DMA, the common activated sludge has low tolerance degree to DMF, and even if the maximum DMF concentration of the domesticated activated sludge tolerance is only 200mg/L, the conventional activated sludge can only treat DMF wastewater with low concentration. The DMF content in industrial wastewater is generally 2000mg/L, so that the excavation of microorganism resources capable of rapidly decomposing and mineralizing DMF is of great significance for the efficient treatment of industrial wastewater with high DMF content.
Paracoccus thiooxidans (Paracoccus sulfuroxidans) is a novel species discovered and named in 2006 by Liu et al (Liu X Y, wang B J, jiang C Y, et al Paracoccus sulfaroxidans sp.nov.a sulfur oxidizer from activated sludge [ J ]. Int J Syst Evol Microbiol,2006,56 (11): 2693-2695.), and few studies have been made on this species so far, and no report has been made on the ability of a strain among Paracoccus thiooxidans peroxydans to degrade DMF.
Disclosure of Invention
In order to solve the technical problems, the invention provides a Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment. The Paracoccus thiooxidans has higher DMF tolerance, and can take DMF as the only carbon source and nitrogen source, thereby realizing efficient ammoniation and mineralization of DMF.
The specific technical scheme of the invention is as follows:
in a first aspect, the invention provides a paracoccus thiooxidans capable of efficiently degrading DMF, wherein the paracoccus thiooxidans is named 175A1-1 and is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) on the 10 th month of 2021, the preserving unit address is the North Chen West Lu No. 1 of the Korean region of Beijing city, the preserving number is CGMCC No.23659, and the microorganism classification is named as paracoccus thiooxidans Paracoccus sulfuroxidans.
The strain 175A1-1 is used for extracting fresh sludge from a pharmaceutical factory, and the 16S rDNA sequence is shown as SEQ ID NO. 1, and belongs to Paracoccus thiooxidans (Paracoccus sulfuroxidans) after identification.
The Paracoccus thiooxidans 175A1-1 has higher DMF tolerance and better DMF degradation capability, and can take DMF as the only carbon source and nitrogen source to realize efficient ammoniation and mineralization of DMF. Through experiments, the concentration of the DMF of the strain can be tolerated to 10000mg/L; and when the DMF concentration is 10000mg/L, the strain can convert DMF into ammonia Nitrogen (NH) 3 -N) efficiency up to 99.0%.
The difficulty in biological treatment of DMF wastewater is that organic nitrogen is difficult to be converted into NH 3 -N. The strain of the invention can degrade DMF into NH with high efficiency 3 and-N, and then combining with a conventional nitrification-denitrification process to realize denitrification of the DMF wastewater.
In a second aspect, the present invention provides a bacterial culture comprising the Paracoccus thiooxidans.
Preferably, the bacterial culture is a bacterial liquid or a bacterial agent.
In a third aspect, the invention provides the use of said s.thiooxidans in the treatment of DMF containing wastewater.
Preferably, the application comprises the steps of:
(1) Performing expansion culture on the paracoccus thiooxidans to obtain seed liquid;
(2) Adding the seed liquid into a DMF degradation device, wherein a filler is arranged in the DMF degradation device;
(3) And (3) introducing the DMF-containing wastewater into a DMF degradation device for DMF degradation treatment.
By adopting the method, the paracoccus thiooxidans 175A1-1 is applied to the actual engineering, and the efficient degradation of DMF in the wastewater can be realized. Through experiments, when the method is adopted to treat the wastewater with the DMF content of 1.2% (w/v), the DMF removal rate in the wastewater can reach more than 99%, and the DMF is completely mineralized and the organic nitrogen is completely aminated into NH 3 -N。
Preferably, the specific process of step (1) is as follows: inoculating the Paracoccus thiooxidans into a culture medium, and culturing until OD 600 The value reaches 1.5-2.0, and the seed liquid is obtained.
Further, the medium comprises the following components in concentration: 1.8-2.3g/L peptone, 0.8-1.2g/L yeast extract, 1.8-2.3g/L NaCl; the pH of the culture medium is 6.0-7.0.
Preferably, in the step (3), the dissolved oxygen content of the wastewater containing DMF in the DMF degradation device is controlled to be more than 2mg/L, the water temperature is 30-35 ℃, and the pH is 6.0-7.0.
Preferably, in the step (2), the seed solution is added into the DMF degradation device according to the inoculation amount of 15-25%, namely, the ratio of the volume of the seed solution to the volume of the wastewater in the DMF degradation device after the seed solution is added is 15-25%.
Preferably, in the step (3), the hydraulic retention time of the DMF containing wastewater in the DMF degradation device is 3-5 days.
Preferably, in the step (2), the filler is activated carbon.
Compared with the prior art, the invention has the following advantages:
(1) The Paracoccus thiooxidans 175A1-1 has higher DMF tolerance, and can convert DMF into NH by taking DMF as the sole carbon source and nitrogen source 3 N, realizing efficient ammoniation and mineralization of DMF, but no report exists in the literature at present that the strain in the paracoccus thioperoxide has the capacity of degrading DMF;
(2) The Paracoccus thiooxidans 175A1-1 can be applied to the field engineering efficiently and stably.
Drawings
FIG. 1 is a graph showing the effect of treatment of COD and TN with strain P.sulfoxidans 175A1-1 at a DMF concentration of 5000mg/L and the effect of ammoniation. Fig. 1 (a) is a graph showing the treatment effect on COD; fig. 1 (B) is a graph showing the effect of processing TN and the effect of ammoniation.
FIG. 2 is a graph showing the effect of treatment of COD and TN with strain A.lucatiense 3-1-1 at a DMF concentration of 5000mg/L and the effect of ammoniation. Wherein, fig. 2 (a) is a graph of treatment effect on COD; fig. 2 (B) is a graph showing the effect of processing TN and the effect of ammoniation.
FIG. 3 is a graph showing the effect of treating COD and TN and the effect of ammoniation by the strain G.flavus 2-CZ-5 at a DMF concentration of 5000mg/L. Wherein, fig. 3 (a) is a graph of treatment effect on COD; fig. 3 (B) is a graph showing the effect of processing TN and the effect of ammoniation.
FIG. 4 is a graph showing the effect of treating COD and TN with strain S.Paucimobilis 3-3-10 at a DMF concentration of 5000mg/L and the effect of ammoniation. Fig. 4 (a) is a graph showing the treatment effect on COD; fig. 4 (B) is a graph showing the effect of processing and ammoniation of TN.
FIG. 5 is a schematic diagram of a continuous water inlet and outlet apparatus for testing the DMF treating effect of the strain in example 3.
FIG. 6 is a graph showing the effect of treatment and ammoniation of COD and TN in a blank group (without adding strain, HRT 3 d) at a DMF concentration of 10000 mg/L. Fig. 6 (a) is a graph showing the effect of COD treatment; fig. 6 (B) is a graph showing the effect of processing and ammoniation of TN.
FIG. 7 is a graph showing the effect of treatment and ammoniation of COD and TN in a control group (activated sludge addition, HRT 3 d) at a DMF concentration of 10000 mg/L. Fig. 7 (a) is a graph showing the effect of COD treatment; fig. 7 (B) is a graph showing the effect of processing and ammoniation of TN.
FIG. 8 is a graph showing the effect of treatment of COD and TN and the effect of ammoniation in the experimental group (P.sulfaroxidans 175A1-1, HRT 3 d) at a DMF concentration of 10000 mg/L. Fig. 8 (a) is a graph showing the effect of COD treatment; fig. 8 (B) is a graph showing the effect of processing and ammoniation of TN.
FIG. 9 is a graph showing the effect of treatment of COD and TN and the effect of ammoniation in the experimental group (P.sulfaroxidans 175A1-1, HRT 5 d) at a DMF concentration of 10000 mg/L. Fig. 9 (a) is a graph showing the effect of COD treatment; fig. 9 (B) is a graph showing the effect of processing and ammoniation of TN.
FIG. 10 is a graph showing the effect of strain P.sulfoxidans 175A1-1 on the treatment of high concentration DMF wastewater. Wherein, fig. 10 (a) is a graph showing the treatment effect of COD in wastewater; fig. 10 (B) is a graph showing the effect of treating TN in wastewater and the effect of ammoniating.
Detailed Description
The invention is further described below with reference to examples.
General examples
The Paracoccus thiooxidans capable of efficiently degrading DMF is named 175A1-1, and is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 10 months and 25 days in 2021, the preservation unit address is North Chen West Lu No. 1, 3 in the Guangxi area of Beijing, the preservation number is CGMCC No.23659, and the microorganism classification is named as Paracoccus thiooxidans Paracoccus sulfuroxidans.
A bacterial culture containing the paracoccus thiooxidans, wherein the bacterial culture is bacterial liquid or bacterial agent.
The method for treating the DMF-containing wastewater by utilizing the paracoccus thiooxidans comprises the following steps:
(1) The culture medium was prepared according to the following formulation: 1.8-2.3g/L peptone, 0.8-1.2g/L yeast extract, and 1.8-2.3g/L NaCl; adjusting the pH to 6.0-7.0;
(2) Inoculating the Paracoccus thiooxidans into a culture medium, and culturing until OD 600 The value reaches 1.5-2.0, and seed liquid is obtained;
(3) The seed liquid is added into a DMF degradation device according to 15-25% of inoculation amount, and the DMF degradation device is internally provided with a filler;
(4) Introducing the DMF-containing wastewater into a DMF degradation device, controlling the dissolved oxygen content of the DMF-containing wastewater in the DMF degradation device to be more than 2mg/L, controlling the water temperature to be 30-35 ℃, controlling the pH to be 6.0-7.0, controlling the hydraulic retention time to be 3-5 days, and carrying out DMF degradation treatment.
Example 1: preparing culture medium
Three media were prepared according to the following formulation:
inorganic salt medium (g/L): KH (KH) 2 PO 4 1.2、K 2 HPO 4 6.8、NaCl 0.5、MgSO 4 ·7H 2 O 0.1、MnSO 4 ·H 2 O 0.1、 CaCl 2 0.1、FeSO 4 ·7H 2 O 0.1、Na 2 MoO 4 ·2H 2 O 0.006、CuSO 4 ·5H 2 O 0.006、ZnSO 4 ·7H 2 O 0.007、 CoCl 2 ·6H 2 O 0.0001、H 3 BO 3 0.0124 and vitamin B1.00001, DMF is used as the only carbon source and nitrogen source, and pH=6 (solid culture medium is added into agar powder with mass fraction of 2%).
LB medium (g/L): peptone 10.0, yeast extract 5.0, naCl 10.0, ph=7 (solid medium added with agar powder 2% by mass).
DMF1 medium: 0.2g/L DMF and 0.2g/L dimethylamine were additionally added to the LB medium.
Example 2: domestication, separation and identification of strains
(1) And (3) strain domestication:
10mL of fresh sludge from each pharmaceutical factory is inoculated into a triangular flask filled with 100mL of inorganic salt culture medium (DMF concentration is 1000 mg/L), and the flask is placed in a shaking incubator at 30 ℃ and 130r/min for culturing for 1 week, and TN and NH in a detection system are sampled every other day during the culture 3 -N value. After 1 week of culture, NH in the system 3 The N is obviously raised to about 160mg/L, and the TN ammoniation rate reaches more than 90 percent. Transferring into fresh inorganic salt culture medium (DMF concentration is 2000 mg/L) according to 10% inoculum size, and monitoring TN and NH in the system 3 N value, further culture for 1 week. The transfer is carried out for 5 times continuously, and the concentration of DMF is gradually increased to 5000mg/L.
Note that: ammoniation refers to the conversion of organic nitrogen to NH in TN 3 -N。
(2) Separating strains:
and 1mL of bacterial liquid in a culture system with good bacterial growth condition is taken for high-throughput sequencing. At the same time press 10 -2 To 10 -8 The bacterial liquid is diluted and coated on an inorganic salt solid culture medium containing 1000mg/L DMF, and the bacterial liquid is subjected to static culture for 5d at 30 ℃. Single colonies were picked up in DMF1 medium, shake-cultured at 30 ℃ for 3d, streak-purified, and isolated into 4 strains.
(3) And (3) strain identification:
the isolated 4 strains were subjected to 16S rDNA sequencing, and the alignment information of the obtained 16S rDNA sequences in the EzBioCloud database is shown in Table 1. Wherein the 16S rDNA sequence of the strain 175A1-1 is shown as SEQ ID NO. 1.
Table 14 16S rDNA sequence alignment information of strains
| Strain numbering | 16S rDNA sequence alignment of most similar strains | Similarity degree |
| 175A1-1 | Paracoccus sulfuroxidans CGMCC 1.5364 T | 99.04％ |
| 3-1-1 | Aquamicrobium lusatiense DSM 11099 T | 100％ |
| 2-CZ-5 | Gelidibacter flavus JCM 31135 T | 100％ |
| 3-3-10 | Sphingomonas paucimobilis NBRC 13935 T | 100％ |
According to the comparison result of the 16S rDNA sequences, the similarity of the strain 175A1-1 and Paracoccus sulfuroxidans CGMCC 1.5364T reaches 99%, a phylogenetic tree is constructed by MEGA software, and the strain 175A1-1 and Paracoccus sulfuroxidans CGMCC 1.5364 are also obtained T Compact phase clusters, therefore identified as Paracoccus sulfuroxidans species.
The characteristics of strain P.sulfoxidans 175A1-1 are:
gram negative bacteria, the length of the cells is 1.0-1.2 μm, the width is 0.5 μm, the short rods are cultivated on LB culture medium for 3d, and the bacterial colony is beige, smooth in edge, round and 0.4-2.0mm in diameter. The temperature growth range is 25-36 ℃ (most suitable for 30-35 ℃), the pH growth range is 5.0-10.0 (most suitable for 6.0-7.0), and the NaCl salinity growth range is 0-5% (most suitable for 0.5%), and glucose, acetic acid, ethanol, citric acid and maltose can be used.
Example 3: ability of single bacteria to degrade DMF
(1) Seed liquid preparation:
the 4 strains P.sulfoxidans 175A1-1, aquamicrobium lusatiense 3-1-1, gelidibacter flavus 2-CZ-5, sphingomonas paucimobilis 3-3-10 isolated in example 2 were grown in LB medium to OD by expansion, respectively 600 All reach about 1.5, centrifuging for 3min at 6000r/min, discarding supernatant, and collecting bottom thallus. Resuspension is carried out by using sterile distilled water to prepare seed liquid.
(2) The experimental device comprises:
the experimental device is a continuous water inlet and outlet device so as to simulate the on-site treatment system to the greatest extent to test the treatment effect of the strain on DMF. The whole set of device is shown in figure 5, and comprises a water inlet tank, a strain treatment tank and a water outlet tank which are connected in sequence. The effective volume of each tank body is 3L, the water inlet tank is used for adding nutrient substances to adjust water quality, the water inlet tank is accommodated, and the bottom of the water inlet tank is provided with a stirring system; the strain treatment tank is a strain biochemical treatment tank, and an aeration system is arranged at the bottom of the strain treatment tank; the wastewater in the water inlet tank is lifted to a strain treatment tank by a peristaltic pump; the water outlet pool is used for collecting the treated wastewater.
(3) And (3) water inlet:
the water inlet in the water inlet tank is an inorganic salt culture medium containing 5000mg/L DMF, the pH is controlled at 6.5+/-0.5, and the wastewater is artificially prepared.
(4) Adding strains:
adding inorganic salt culture medium into the strain treatment tank, controlling pH at 6.5+ -0.5, inoculating each seed solution into the strain treatment tank according to 20% of inoculation amount, and adding granular activated carbon as strain carrier to enrich and entrap strain.
(5) And (3) process control:
controlling HRT (hydraulic retention time) of inlet water in a strain treatment tank to be 3d and DO>2.0mg/L, water temperature is 30-35 ℃. During the strain treatment, the pH of the strain treatment tank can rise to 9.0, H needs to be added 2 SO 4 The pH of the strain treatment pool is always controlled to be=6.0-7.0.
(6) And (3) data monitoring:
basic water quality data (including COD and NH) in the system is monitored every day 3 N and TN), the results are shown in fig. 1-4. The average value of 7-14d was calculated and the results are shown in Table 2.
Table 24 the results of the strain on the treatment of various indexes of wastewater are summarized
1 The calculation formula of the ammoniation rate is as follows: ammoniation rate= (NH in seed treatment tank 3 -N concentration)/(TN concentration in the seed treatment cell).
When 4 strains are tested, the strain has good growth condition, large bacterial load and strong activity. The pH in the strain treatment tank is obviously raised to above 8.0, and a certain amount of acid is required to be added every day, so that the pH is regulated to about 6.
As can be seen from fig. 1-4 and table 2:
DMF degradation effect of P.sulfoxidans 175A1-1 is superior to that of other 3 single bacteria. In the experimental system of P.sulfoxidans 175A1-1, NH was measured in the first 6d time period 3 N and TN both show a slow rising trend, NH 3 The proportion of N to TN is substantially above 95%; NH in the treatment System after 7d 3 The data of-N and TN are basically stable at 780mg/L or so, NH 3 The proportion of N to TN is maintained above 98%, and the DMF degradation effect is obvious.
In the lusatiense 3-1-1 experiment system, NH in the treatment system after 7d 3 The N data is basically stabilized at about 575mg/L, the TN data is stabilized at about 780mg/L, and the NH is stabilized at about 3 The proportion of N to TN is maintained at about 74%.
In the G.flavus 2-CZ-5 experiment system, NH in the treatment system after 7d 3 The N data is basically stabilized at about 473mg/L, the TN data is stabilized at about 780mg/L, and the NH is stabilized at about 3 The proportion of N to TN is maintained at about 60%.
S. Paucimobilis 3-3-10 experiment System, NH in the treatment System after 7d 3 The N data is basically stable at about 572mg/L, the TN data is about 780mg/L, and the NH is basically stable at about 3 The proportion of N to TN is maintained at about 73%.
Since the treatment effect of P.sulfoxidans 175A1-1 is obvious, the subsequent study is focused on.
Example 4: the ability of the strain Paracoccus sulfuroxidans A1-1 to treat DMF at high concentration was further confirmed by the strain P.sulfoxidans 175A1-1 to treat DMF at high concentration, and the concentration of DMF in the influent water was increased to 10000mg/L, and the procedure was the same as in example 3. The system without adding strain was set as a blank group, the system with adding strain P.sulfoxidans 175A1-1 seed solution (prepared in example 3) was set as an experimental group, the system with adding activated sludge (MLVSS is 6000 mg/L) was set as a control group, and the treatment effects of HRT of 3d and 5d in the strain treatment tank were explored. The aim was to look at the resistance of strain P.sulfoxidans 175A1-1 to DMF and compare the effect of this strain with the DMF treatment of activated sludge.
The basic water quality data in the system were monitored daily and the results are shown in figures 6-9. The average value of 7-14d was calculated and the results are shown in Table 3.
Table 3 P.sulfuroxidans 175A1-1 summary of the effects of high concentration DMF
1 The calculation formula of the ammoniation rate is as follows: ammoniation rate= (NH in seed treatment tank 3 -N concentration)/(TN concentration in the seed treatment cell).
As can be seen from fig. 6-9 and table 3:
when the DMF concentration of the inlet water reaches 10000mg/L, the control group (adding activated sludge) can be poisoned, the later period of ammoniation is reduced, and the final ammoniation rate is as low as 14.6%. The experimental group (P.sulfoxidans 175A1-1 strain is added) has obvious growth of thalli in a treatment system, better activity and higher COD removal capacity and ammoniation capacity, and when the HRT is 3d, the COD removal rate is 91.0 percent and the ammoniation rate is 80.0 percent; when the HRT is prolonged to 5d, the COD removal rate is continuously increased to 95.9%, and the ammoniation rate can reach 99.0%.
Namely P.sulfoxidans 175A1-1 tolerating DMF up to 10000mgL and when the HRT is controlled at 5d, the organic nitrogen in DMF is converted to NH 3 The efficiency of N can reach 99.0%.
Example 5: ability of strain Paracoccus sulfuroxidans 175A1-1 to treat DMF wastewater
(1) Waste water source:
the high concentration DMF wastewater from some pharmaceutical industry in Zhejiang river has DMF content of about 1.2% (w/v), water amount of 60t/d, and wastewater quality as shown in Table 4.
TABLE 4 basic Water quality of high concentration DMF wastewater
(2) Strain preparation:
in order to reduce the introduction of nitrogenous substances into wastewater of a strain culture medium, and no centrifugal equipment is used in practical engineering, the strain is fermented by adopting an LB culture medium with 1/5 concentration (namely, the formula is 2.0g/L peptone, 1.0g/L yeast extract, 2.0g/L NaCl and pH of 7.0) until reaching OD 600 When the value reaches about 1.5, seed liquid is obtained.
(3) And (3) regulating nutrition of the wastewater:
KH is supplemented in the waste water 2 PO 4 0.5g/L, and H is used 2 SO 4 Wastewater ph=6.0 was adjusted.
(3) Continuous water inlet and outlet biological treatment:
the continuous water inlet and outlet pool is utilized, a filler layer is arranged at the position 1/3 of the pool from the bottom, the filler is granular activated carbon, and the adding amount is 10% (the filler occupies the volume ratio of the pool).
The aerobic process is adopted to control the HRT to about 5d and DO>2mg/L, water temperature of 30-33 ℃, and P.sulfoxidans 175A1-1 seed solution according to the inoculation amount of 20 percent. During the strain treatment, the pH of the strain treatment tank can rise to 9.0, H needs to be added 2 SO 4 The pH of the strain treatment pool is always controlled to be=6.0-7.0.
The basic water quality data in the system were monitored daily and the results are shown in figure 10. The average value of 6-30d was calculated and the results are shown in Table 5.
Table 5 P.sulfuroxidans 175A1-1 summary of the effects of high concentration DMF wastewater treatment
1 The calculation formula of the ammoniation rate is as follows: ammoniation rate= (NH in seed treatment tank 3 -N concentration)/(TN concentration in the seed treatment cell).
As can be seen from fig. 10 and table 5:
p.sulfoxidans 175A1-1 strain has good wastewater treatment effect on DMF content reaching 1.2% (w/v) and TN of about 2000mg/L, and the COD removal rate is about 95%, thus mineralization can be basically realized; the DMF removal rate reaches 100%. The residual COD is presumed to be an intermediate product in the DMF degradation process by combining with the COD data; substantially all TN conversion to NH 3 The ammoniation rate reaches 99.89%, the treated wastewater is basically free of organic nitrogen, and the denitrification can be realized by carrying out the traditional nitrification and denitrification process.
The raw materials and equipment used in the invention are common raw materials and equipment in the field unless specified otherwise; the methods used in the present invention are conventional in the art unless otherwise specified.
The foregoing description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and any simple modification, variation and equivalent transformation of the above embodiment according to the technical substance of the present invention still fall within the scope of the technical solution of the present invention.
Sequence listing
<110> Xiuchuang science and technology Co., ltd
<120> Paracoccus thiooxidans capable of efficiently degrading DMF and application thereof in DMF-containing wastewater treatment
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<213> Paracoccus thiooxidans 175A1-1 (Paracoccus sulfuroxidans 175A 1-1)
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aacgcgtggg aatatgccct tctctacgga atagtcctgg gaaactgggg gtaataccgt 120
atacgcccta cgggggaaag atttatcgga gaaggattag cccgcgttgg attagctagt 180
tggtgaggta acggctcacc aaggcgacga tccatagctg gtttgagagg atgatcagcc 240
acactgggac tgagacacgg cccagactcc tacgggaggc agcagtgggg aatcttagac 300
aatgggggaa accctgatct agccatgccg cgtgagtgat gaaggcctta gggttgtaaa 360
gctctttcag ctgggaagat aatgacggta ccagcagaag aagccccggc taactccgtg 420
ccagcagccg cggtaatacg gagggggcta gcgttgttcg gaattactgg gcgtaaagcg 480
cacgtaggcg gactggaaag ttggaggtga aatcctgggg ctcaacccca gaactgcctt 540
caaaactatc agtctggagt tcgagagagg tgagtggaat tccgagtgta gaggtgaaat 600
tcgtagatat tcggaggaac accagtggcg aaggcggctc actggctcga tactgacgct 660
gaggtgcgaa agcgtgggga gcaaacagga ttagataccc tggtagtcca cgccgtaaac 720
gatgaatgcc agtcgtcggg tagcatgcta ttcggtgaca cacctaacgg attaagcatt 780
ccgcctgggg agtacggtcg caagattaaa actcaaagga attgacgggg gcccgcacaa 840
gcggtggagc atgtggttta attcgaagca acgcgcagaa ccttaccaac ccttgacatc 900
tagtgctaca tccagagatg gatggttccc ttcggggacg ctaagacagg tgctgcatgg 960
ctgtcgtcag ctcgtgtcgt gagatgttcg gttaagtccg gcaacgagcg caacccacac 1020
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aggtgtggat gacgtcaagt cctcatggcc cttacgggtt gggctacaca cgtgctacaa 1140
tggtggtgac agtgggttaa tccccaaaag ccatctcagt tcggattggg gtctgcaact 1200
cgaccccatg aagttggaat cgctagtaat cgcggaacag catgccgcgg tgaatacgtt 1260
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cgctaacctt tggaggcagc gaccacgtag atacgc 1356
Claims (9)
1. A strain of paracoccus thiooxidans capable of efficiently degrading DMF is characterized in that the paracoccus thiooxidans is named 175A1-1 and is preserved in China general microbiological culture collection center (CGMCC) No.23659 in the 10 th month 25 th year 2021, and the microorganism classification is named paracoccus thiooxidansParacoccus sulfuroxidans。
2. A bacterial culture comprising the paracoccus thiooxidans of claim 1, wherein the bacterial culture is a bacterial liquid or a bacterial agent.
3. Use of paracoccus thiooxidans according to claim 1 in the treatment of DMF containing wastewater.
4. The use according to claim 3, comprising the steps of:
(1) Performing expansion culture on the paracoccus thiooxidans to obtain seed liquid;
(2) Adding the seed liquid into a DMF degradation device, wherein a filler is arranged in the DMF degradation device;
(3) And (3) introducing the DMF-containing wastewater into a DMF degradation device for DMF degradation treatment.
5. The use of claim 4, wherein the procedure of step (1) is as follows: inoculating the Paracoccus thiooxidans into a culture medium, and culturing until OD 600 The value reaches 1.5-2.0, and the seed liquid is obtained.
6. The use according to claim 5, wherein the medium comprises the following concentrations of components: 1.8-2.3g/L peptone, 0.8-1.2g/L yeast extract, 1.8-2.3g/L NaCl; the pH of the culture medium is 6.0-7.0.
7. The use according to claim 4, wherein in step (3), the DMF-containing wastewater in the DMF degradation device is controlled to have a dissolved oxygen content of more than 2mg/L, a water temperature of 30-35 ℃ and a pH of 6.0-7.0.
8. The use according to claim 4, wherein in step (2) the seed liquid is fed to the DMF degradation apparatus in an inoculum size of 15-25%.
9. The use according to claim 4, wherein in step (3), the hydraulic retention time of the DMF containing wastewater in the DMF degradation apparatus is 3-5 days.
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| CN110656057A (en) * | 2018-06-29 | 2020-01-07 | 龙岩学院 | Heterotrophic nitrification-aerobic denitrification paracoccus strain, seed liquid, preparation method and application thereof |
| CN110656071A (en) * | 2019-11-11 | 2020-01-07 | 黄河三角洲京博化工研究院有限公司 | Paracoccus huilkii for efficiently degrading DMF (dimethyl formamide) and application thereof |
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