High-salinity-tolerance bacterium and application thereof
Technical Field
The invention relates to a high-salinity-tolerance bacterium and application thereof in removing COD (chemical oxygen demand) from high-salinity wastewater, belonging to the technical field of microorganisms and wastewater treatment.
Background
High-salinity wastewater, especially high-sulfate organic wastewater, is easily generated in the industries of chemical industry, pharmacy, papermaking, food processing, mining, fermentation and the like, the concentration of sulfate in some wastewater can reach 30000-50000 mg/L, and COD (chemical oxygen demand) exceeds 10000 mg/L. Due to the large amount of sulfate ions, the high-salt-content wastewater is not easy to be biochemically generated, and due to the high organic matters, the physicochemical process routes such as membrane separation, concentration and crystallization are not economical and are not advisable (easy to be blocked).
For such waste water, the existing methods and research directions are as follows:
(1) chemical method, i.e. adding lime to convert sulfate into calcium sulfate precipitate. For example, in patents CN106865880A and CN105439374A, the core of sulfate removal is lime neutralization and chemical flocculation is adopted, and if the process is also adopted for high COD organic wastewater, calcium sulfate precipitation is generated when lime and flocculant are added, and a large amount of organic matters and heavy metals are mixed into the precipitation, so that finally generated lime slag and sludge can only be used as hazardous waste, and have no utility value and secondary pollution.
(2) Simple biochemical method, namely adopting a first-stage anaerobic process. The anaerobic process has higher volume load than the aerobic process, so it is mostly adoptedAnaerobic treatment of organic waste water. If the waste water contains sulfate, the sulfate is reduced to S under the action of Sulfate Reducing Bacteria (SRB) under anaerobic condition2-The ions have stronger biological toxicity, have larger inhibiting effect on microbial flora, particularly methanogen, and seriously influence the removal of organic matters. Therefore in order to reduce S2-The influence of ions on methanogens generally requires that the sulfate radical concentration of the primary anaerobic reactor is less than 2000mg/L, but the method is not suitable for organic wastewater with sulfate radical concentration more than 10000 mg/L. For example, CN103771670A patent, the sulfate radical concentration is below 1000mg/L, and a single anaerobic and aerobic process is adopted, but the process is not suitable for waste water of binary acid fermentation process.
(3) Two-stage anaerobic process, i.e. to avoid the mutual competition of sulfate reducing bacteria and methanogens in the anaerobic process, at present, two-stage anaerobic process is mostly adopted at home and abroad to treat high-concentration organic wastewater. As described in patent CN105439374A, the primary anaerobic control is in hydrolysis acidification stage, and sulfate reducing bacteria reduce most of sulfate in wastewater to S2-Ion and with H+Hydrogen sulfide is generated by combination, and enters a dry desulfurizer for desulfurization after being blown off by nitrogen; and the second-stage anaerobic control is in a methane production stage, and anaerobic effluent enters an aerobic reaction tank for further treatment. The dry desulfurization used in the first-stage anaerobic section of the patent is a relatively old technology, has great defects in desulfurizer replacement, continuous operation, regeneration treatment and the like, and does not make clear the final destination of the desulfurized sulfur; the organic matter in the secondary anaerobic process finally generates methane, carbon dioxide, water and a small amount of hydrogen sulfide, namely methane, and the treatment of the methane is also ignored in the patent.
(4) Biological desulfurization technique, i.e. sulfate radical is reduced into S by sulfate reducing bacteria under anaerobic condition2-Or hydrogen sulfide, and then biologically oxidized by sulfur oxidizing bacteria to generate elemental sulfur, such as patents CN102795739A, CN103172218A, and CN 103319002A. The biological desulfurization technology has the defects of difficult process control and strict condition requirement, and the problems of poor separation effect and low sulfur purity in the liquid-phase sulfur preparation process, so the technology is developed for decades but is separated from industrial application at presentBut also a certain distance.
(5) The special halotolerant bacteria biotechnology is based on the rapid development of genetic engineering technology, and through a scientific method, dominant flora suitable for high-salinity wastewater is domesticated, and the thalli can grow in an environment with higher salinity by virtue of the unique cell structure and material composition of the thalli, such as patents CN201610547861, CN201510626828, CN201610720403 and CN 201510737150. At present, NaCl is mostly used as a main component of salt in strain screening and culture in the patent in the direction, and the mass ratio of sulfate is low because S is generated due to local anaerobic reaction when the concentration of sulfate in a culture medium/liquid is high2-Or hydrogen sulfide, to bring some biological toxicity, inhibit or kill the bacterial species. Moreover, the halotolerant bacteria related to the patent only simply states that the halotolerant bacteria can be used in high-salinity wastewater, the stability and long-period adaptability of the strains in a special environment are not investigated, specific implementation details are also lacked, and the salinity wastewater is NaCl-containing wastewater.
Disclosure of Invention
Aiming at the defects, the invention provides a strain of high-salinity-tolerance bacteria and application thereof in removing COD in high-salinity wastewater, and has the characteristics of high-salinity resistance, especially high-sulfate resistance.
The technical purpose of the invention is realized by the following technical scheme:
the technical purpose of the first aspect of the invention is to provide a strain of high-salt-tolerance bacteria (Halomonas nigrificans) GXNYJ-DL-1 is preserved in China general microbiological culture Collection center (CGMCC) at 13.7.2020, with the preservation number of CGMCC 20350.
The high salt-tolerant bacteria GXNYJ-DL-1 provided by the invention has the following forms: the bacterial colony is light yellow and round, the surface is wet and opaque, and the edge is neat; microscopic morphology under microscope was: the bacteria are rod-shaped, 0.5-0.7 μm multiplied by 1.2-3.6 μm, arranged singly or in pairs and gram-negative.
The sequencing analysis result of the 16S rDNA gene of the high-salinity-tolerant bacteria GXNYJ-DL-1 provided by the invention is shown in a sequence table.
The technical purpose of the second aspect of the invention is to provide a culture method of the high-salinity-tolerant bacterium GXNYJ-DL-1, which comprises three stages of strain activation, seed solution culture and intermittent aeration culture, and comprises the following specific steps:
(1) strain activation: inoculating the strain GXNYJ-DL-1 into a broth peptone solid medium with the salt content of 1-5% by adopting a plate streaking method, and culturing in an incubator at 28-35 ℃ for 48-72 h.
(2) Seed liquid culture: after the activation of the strains is finished, selecting the activated strains in the flat plate, inoculating the strains into a conical flask containing a broth peptone liquid culture medium, wherein the liquid culture medium contains 1-5% of salt and more than 50% of sulfate, performing shake culture on a shaking table at the temperature of 30-35 ℃, the rotation speed of 100-200 rpm, and the culture time of 24-96 h.
(3) Intermittent aeration culture: adding the prepared salt-containing wastewater or the salt-containing wastewater to be treated, wherein the salt content is 1-5%, the sulfate accounts for more than 50%, inoculating liquid seed liquid according to the volume ratio of 3-20%, controlling the pH value to be 6-9, carrying out indirect aeration according to the relationship between the aeration duration and the aeration stop duration 2: 1-1: 2, and the total culture period is 72-144 h; s production by sulfate reduction reaction upon indirect aeration2-Improving the S resistance of the strain GXNYJ-DL-12-And (4) capability of preventing other mixed bacteria from breeding.
The high-salt-tolerance GXNYJ-DL-1 bacterial liquid obtained by the culture method provided by the invention can be kept still in a refrigerator at 4 ℃ for 3-5 months without inactivation, and has strong vitality and high stability.
The technical purpose of the third aspect of the invention is to provide the application of the high salt-tolerant bacteria GXNYJ-DL-1 in removing COD in high salt organic wastewater.
Furthermore, the high-salinity-tolerant bacteria GXNYJ-DL-1 can tolerate salt up to 25wt% and S2-The concentration reaches 300mg/L, and the method is more suitable for treating high-salinity wastewater, particularly the saline wastewater with sulfate as a main body compared with other salt-tolerant bacteria.
The technical purpose of the fourth aspect of the invention is to provide a process for treating high-sulfate organic wastewater by using the high-salt-tolerant bacteria GXNYJ-DL-1, which comprises a front-end treatment section, an advanced treatment section, a sludge treatment section and a tail gas treatment section.
The front-end treatment section sequentially comprises pH adjustment, primary aerobic treatment, anaerobic treatment, stripping treatment and secondary aerobic treatment; the pH adjustment is to add a pH regulator into the wastewater to adjust the pH of the wastewater to 6.5-7.5, then to perform primary aerobic treatment, and to treat the high-sulfate organic wastewater by using high-salt-tolerant bacteria GXNYJ-DL-1 to reduce the COD of the high-sulfate organic wastewater to below 1000 mg/L; the primary aerobic effluent enters an anaerobic section, sulfate is reduced into hydrogen sulfide under the action of sulfate reducing bacteria, and simultaneously, methane is used as stripping gas for stripping, so that the generated hydrogen sulfide is taken out of a wastewater system; and the anaerobic effluent enters a secondary aerobic treatment, the COD is further removed by adopting conventional sludge strains, and the secondary aerobic effluent enters an advanced treatment section.
The advanced treatment section sequentially comprises advanced oxidation, BAF and postposition dephosphorization; the advanced oxidation is to remove refractory organic matters in the wastewater, improve the biodegradability of the wastewater, further remove COD in the wastewater through BAF and play a role in filtering; the post-dephosphorization is to carry out two-stage chemical dephosphorization by adding medicament, and finally obtain the wastewater meeting the discharge standard.
And in the sludge treatment section, redundant sludge generated by the primary aerobic unit, the secondary aerobic unit and the BAF unit is conveyed to the sludge anaerobic unit by a pump, and most of activated sludge is converted into methane gas through sludge anaerobic oxidation and is recycled to the methane storage tank.
The tail gas treatment section comprises solvent absorption, solvent regeneration and a WSA wet method for preparing sulfuric acid; the solvent absorption is to absorb hydrogen sulfide gas in the gas by an absorbent, purify the gas to obtain methane and store the methane in a methane storage tank; and the solvent regeneration is to heat the gas absorbed by the solvent to escape, the solvent is recycled, and the escaped gas is used for preparing sulfuric acid by a WSA wet method to obtain a sulfuric acid product.
Further, the pH adjusting agent used for pH adjustment is an alkali which does not precipitate with sulfate radicals or slightly dissolve compounds after being added to the wastewater, more specifically, sodium hydroxide or potassium hydroxide, and the pH adjustment is performed in an adjusting tank.
Further, the high-salinity-tolerance bacterium GXNYJ-DL-1 is adopted in the primary aerobic process and is preferably selected from the processes with high volume loads such as a biological contact oxidation process, an MBBR process and the likeOf 2kg (BOD) in volume5)/m3D or more, controlling the dissolved oxygen to be more than 2mg/L, and keeping the waste water for 24-120 h.
Further, dissolved oxygen in the anaerobic treatment wastewater is controlled to be below 0.2mg/L, methane gas is used for stripping, the retention time of the wastewater is 12-144 h, the temperature is 25-35 ℃, and the volume ratio of the methane stripping gas to the wastewater is not less than 5: 1.
As will be understood by those skilled in the art, the primary aerobic section can decompose most organic matters in the wastewater by the high salt-tolerant bacteria GXNYJ-DL-1, partially convert the organic matters into inorganic carbon (carbon dioxide), partially transfer the inorganic carbon into activated sludge in the form of organic carbon, and remove the organic carbon by sludge discharge; the high-efficiency halotolerant strain GXNYJ-DL-1 solves the problem that common strains cannot survive under the condition of high salt content, and also solves the problem that common halotolerant strains can not survive due to nonuniform aeration or local anaerobic oxidation of flora under the condition of a large amount of sulfate2-High concentration and even no survival; the anaerobic section is a sulfate reduction section, most of sulfate is reduced into hydrogen sulfide under the action of sulfate reducing bacteria, redundant hydrogen sulfide gas is taken out of the system through continuous methane stripping, and because most of organic matters are removed in the primary aerobic section, a small amount of residual organic matters are not enough to enable methanogenic bacteria to propagate in a large amount, the sulfate reducing bacteria are taken as a main body at the moment, and a small amount of organic matters are consumed as a carbon source of the sulfate reducing bacteria.
Further, the secondary aerobic treatment adopts a conventional activated sludge method and uses conventional biological flora, and the retention time of the wastewater is 12-48 h; the secondary aerobic treatment mainly treats the primary aerobic residual organic matters and the partial acidification hydrolysis organic matters in the anaerobic section, and a large amount of organic matters and sulfate are removed in the unit, so that the unit has moderate salt concentration and volume load, the treatment effect can be achieved by conventional biological flora in a short time, and the treatment pressure and cost of a subsequent advanced oxidation unit are reduced.
Further, the advanced oxidation is selected from one of ozone oxidation, electrocatalytic oxidation and fenton oxidation. To decompose organic substances which are difficult to degrade in the wastewater and improve the biodegradability of the wastewater; the BAF can further remove COD from the wastewater and act as a filter, and the BAF unit can also be replaced with a similarly acting MBR.
Furthermore, two-stage chemical phosphorus removal is adopted for the post-phosphorus removal, and the chemical agent is a ferric chloride and calcium hydroxide composite agent and mainly aims at the problem that the total phosphorus content in the dibasic acid fermentation wastewater is relatively high; the ferric chloride is acidic, the calcium hydroxide is alkaline, the ferric chloride and the calcium hydroxide are compounded according to a certain proportion to ensure that the pH of the liquid is neutral, and the subsequent effluent does not need to be subjected to pH adjustment; the dosage of the first-grade phosphorus removal ferric chloride is 34 mg/L-680 mg/L, and the dosage of the calcium hydroxide is 10 mg/L-300 mg/L; the dosage of the secondary phosphorus removal ferric chloride is 10 mg/L-200 mg/L, and the dosage of the calcium hydroxide is 4 mg/L-80 mg/L; the iron phosphate and calcium phosphate generated by the postpositive dephosphorization are filtered and recovered and can be used as phosphate fertilizer.
Furthermore, the sludge treatment section mainly recovers sludge generated in the aerobic section, biological sludge is converted into methane through anaerobic oxidation of the sludge, and the primary aerobic section can generate a large amount of biological sludge in the process of removing organic matters, so that the sludge treatment section can effectively recover the resources of the organic matters.
Further, the absorbent of the tail gas treatment section is selected from one of monoethanolamine, diethanolamine, diisopropanolamine and N-methyldiethanolamine, and is preferably N-methyldiethanolamine. The absorbent absorbs primarily hydrogen sulfide gas, including small amounts of carbon dioxide.
Further, the solvent regeneration is carried out in a heated distillation column.
Furthermore, the purity of the methane in the methane storage tank can reach more than 90% after the methane is absorbed and separated by the preorder solvent, wherein part of the methane is recycled outside the stripping gas, and the redundant part of the methane can be used as a product for resource utilization.
Further, the WSA wet process sulfuric acid is to prepare sulfuric acid through incineration, conversion and condensation processes: the gas separated from the solvent regeneration is incinerated, wherein H is2Combustion of S to SO2,SO2Conversion to SO over a catalyst3,SO3And the water vapor enters a condenser to be condensed to generate sulfuric acid.
Compared with the prior art, the invention has the following advantages:
(1) the high-salt-tolerance strain GXNYJ-DL-1 is obtained by screening and culturing, and the culture method is provided, so that the GXNYJ-DL-1 bacterial liquid obtained by the culture method can be stored at 0-8 ℃ for 3-5 months without inactivation, and has very strong stability. The high-salt-tolerance strain GXNYJ-DL-1 has excellent salt tolerance, can particularly tolerate high sulfate, and is more suitable for removing COD (chemical oxygen demand) of high-salt wastewater, particularly high-sulfate wastewater compared with the high-salt-tolerance strain in the prior art.
(2) Aiming at the characteristic that the sulfate radical source of the wastewater is sulfuric acid added in the acidification fermentation process, the invention also provides a process route which takes a high salt-tolerant strain GXNYJ-DL-1 as a core and aims at recycling resources, thereby not only solving the problem of overhigh sulfate radical and COD in the wastewater, but also providing a process raw material (sulfuric acid) for the source and simultaneously realizing the resource utilization of organic matters-biological sludge-methane.
(3) The method for treating the binary acid fermentation sewage realizes the maximization of the resource recycling of the sulfur element, and is specifically embodied in that: sodium hydroxide, potassium hydroxide and the like are added into the pH adjusting tank instead of calcium hydroxide, so that sulfur does not enter solid hazardous waste mainly containing calcium sulfate and flocculating agent; maximizing sulfate reduction with independent anaerobism; the recovery of hydrogen sulfide is enhanced by methane stripping; the recycling of sulfuric acid and the sulfur recovery are realized by combining solvent absorption, regeneration and WSA wet-process acid preparation, and finally, the high-efficiency recovery of hydrogen sulfide and the cyclic utilization of sulfur element are realized.
(4) The dibasic acid fermentation sewage treatment process provided by the invention develops process routes of sludge recovery, sludge anaerobic oxidation, methane circulation stripping, methane separation and purification and methane recovery and utilization aiming at the characteristic of high sludge yield of an aerobic process, realizes resource utilization of organic matters, and simultaneously prepares high-purity methane.
(5) The dibasic acid fermentation sewage treatment process finally realizes the standard discharge of the waste water, the used postposition two-stage phosphorus removal can realize the recycling of phosphate fertilizer while removing the total phosphorus in the sewage, and simultaneously, the hydrogen sulfide and the methane are recycled, so that the whole process greatly reduces the secondary pollution.
Drawings
FIG. 1 is a microscopic morphological diagram of a high salt-tolerant strain GXNYJ-DL-1 under a microscope;
FIG. 2 is a colony morphology picture of high salt-tolerant strain GXNYJ-DL-1 on solid culture medium;
FIG. 3 growth curves of the strains in example 3 at different salt concentrations;
FIG. 4. removal rate of COD by the strain in example 3 at different salt concentrations;
FIG. 5. bacterial strain in example 4 at S2-Growth curve at concentration;
FIG. 6 is a flow chart of the treatment of sewage by using long-chain dicarboxylic acid fermentation according to the present invention.
Biological material preservation instructions
The high salt-tolerant strain (A) provided by the inventionHalomonas nigrificans) GXNYJ-DL-1, deposited in the China general microbiological culture Collection center; address: the institute of microbiology, national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, Beijing; the preservation number is: CGMCC number 20350; the preservation date is as follows: year 2020, 7, 13.
Detailed Description
The present invention will be described in further detail with reference to specific examples. The embodiments are carried out on the premise of the technical scheme of the invention, and detailed embodiments and specific operation processes are given, but the scope of the invention is not limited to the following embodiments.
Example 1
The screening process of the high-salt-tolerant strain GXNYJ-DL-1 provided by the invention is as follows:
(1) sampling: activated sludge and sewage samples of a Qingjiang petrochemical binary acid sewage treatment plant in Huaian city, Jiangsu province are taken and stored in a refrigerator at 4 ℃ for later use.
(2) Sludge activation: and (3) taking the standby sewage and activated sludge in the refrigerator, putting the standby sewage and the activated sludge into a reactor provided with an aeration device, and carrying out aeration for 48-72 hours while controlling the dissolved oxygen to be more than or equal to 2 mg/L.
(3) Primary domestication: after the aeration, 10mL of mixed bacterial liquid is taken and put into a conical flask filled with 200mL of liquid culture medium, then the conical flask is put into an oscillator for culture, the temperature is 30-35 ℃, the rotating speed is 100-200 rpm, the culture period is 48-72 h, after the culture is finished, 10mL of bacterial liquid is taken and put into the liquid culture medium of the second batch, and so on; the salt content of the first liquid culture medium is 1% (mass percentage concentration), the second is 2%, the fifth is 5%, and 5 batches are total; the liquid culture medium is a broth peptone culture medium, and comprises the following components: 3g of beef extract, 10g of peptone, 1000ml of distilled water, 5g of NaCl, pH 7 and Na2SO45-45 g, and preparing liquid culture media with different mass concentrations by using different sodium sulfate proportions.
(4) And (3) domesticating again: after the primary acclimatization is finished, 10ml of the bacterial liquid of the 5 th batch is taken and added into a conical flask filled with 200ml of liquid culture medium, the salt content of the liquid culture medium is 5 percent, and then sodium sulfide is added to prepare S2-The concentration is 100mg/L, then the mixture is put into an oscillator for culture, the culture period is 48-72 h, 10ml of bacterial liquid is taken out after the culture is finished and is put into a liquid culture medium of a second batch, and so on; second liquid Medium S2-The concentration was 150mg/L, the fifth 300%, for a total of 5 batches.
(5) Screening high-efficiency salt-tolerant strains: taking out the salt content of 5 percent and S2-Diluting 1mL of bacterial liquid in a liquid culture medium with the initial concentration of 300mg/L and after shaking reaction for 48-72 h by 1000 times with sterile distilled water, then inoculating the bacterial liquid into a solid culture medium by adopting a flat plate streaking separation method, and then culturing the bacterial liquid for 48-72 h at 30 ℃; after the culture is finished, selecting strains with different colony forms, continuously inoculating the strains into a new solid culture medium by adopting a plate streaking separation method, repeating for 3-8 times until strains with consistent colony forms are screened out, and finally storing in a refrigerator at 4 ℃; the solid culture medium is broth peptone culture medium added with 20g of agar, wherein Na2SO4Was 45 g.
The form of the strain under a microscope is that the strain is rod-shaped, 0.5-0.7 mu m multiplied by 1.2-3.6 mu m, arranged singly or in pairs and gram-negative. The solid culture colonies were pale yellow, round, wet in surface, opaque, and with clean edges, as shown in FIGS. 1 and 2.
Example 2
Identification of the strain:
through physiological and biochemical identification and 16S rDNA gene sequencing analysis, the physiological and biochemical identification results are shown in a table 1, and the gene sequence determination results are shown in a sequence table.
TABLE 1
Note: "+" indicates a positive reaction or availability; "-" indicates a negative reaction or no utilization.
Example 3
Determination of salt tolerance of high salt-tolerant bacteria GXNYJ-DL-1
Preparing simulated wastewater (g/L): phenol 0.4, NaCl 3, FeSO4 0.02,CaCl2 0.03,MgSO 4 1,Na2SO43,KH2PO4 0.034 ,NH4Cl 0.3, yeast extract 0.1, tryptone 0.05g, pH 7, salt content about 1% (mass percent). Additionally adding Na on the basis of 1 percent of salt content simulation wastewater2SO4Preparing the wastewater with salt contents of 5%, 9%, 13%, 17%, 21% and 25% respectively.
Adding GXNYJ-DL-1 bacterial liquid into a conical flask according to the volume ratio of the GXNYJ-DL-1 bacterial liquid to the simulated wastewater of 1:20, adopting a shaking table oscillation method, controlling the temperature at 35 ℃, rotating at 150rpm, sampling at fixed time, and measuring the bacterial density (OD) by using a spectrophotometer600) Drawing a strain growth curve, wherein the strain growth curve is shown in figure 3 under different salt concentrations; and the COD value of the final reaction solution was measured to determine the removal rate of COD by the strain, and FIG. 4 shows the removal rate of COD by the strain after 76h at different salt concentrations.
As can be seen from the graphs in FIGS. 3 and 4, the growth of the strain is relatively slowed down with the increase of the salt concentration, but the strain can rapidly grow after a certain adaptation period, the strain grows faster under the salt concentration of 1% -13%, and the COD removal rate (initial phenol COD is about 1247 mg/L) is up to more than 65%; at 25% salt concentrationThe adaptation period of the strain is relatively long, about 50h, and then the strain begins to enter the growth period, OD600The value is obviously increased, and the removal rate of COD can still reach 53 percent.
According to the embodiment, the strain GXNYJ-DL-1 has stronger salt tolerance, and the COD removal rate can still reach 53% under the condition that the salt concentration is 25%.
Example 4
S-resistant of high-salt-tolerant bacterium GXNYJ-DL-12-Toxicity assay
Preparing simulated wastewater (g/L): phenol 0.4, NaCl 3, FeSO4 0.02,CaCl2 0.03,MgSO 4 1,Na2SO443,KH2PO4 0.034 ,NH4Cl 0.3, yeast extract 0.1, tryptone 0.05g, pH 7, salt content about 5% (mass percent). Additionally adding Na on the basis of simulated wastewater2S is prepared into S2-Wastewater with mass concentrations of 0mg/L, 50mg/L, 100mg/L, 150mg/L, 200mg/L, 250mg/L and 300 mg/L.
Adding GXNYJ-DL-1 bacterial liquid into a conical flask according to the volume ratio of the GXNYJ-DL-1 bacterial liquid to the simulated wastewater of 1:20, standing for 24h, shaking by using a shaking table, controlling the temperature at 35 ℃, rotating at 150rpm, sampling at regular time, and measuring the bacterial density (OD) by using a spectrophotometer600) The strain growth curves were plotted, as shown in FIG. 5.
As can be seen from FIG. 5, the growth of the strain was very slow during the standing, limited on the one hand by dissolved oxygen and on the other hand by S2-Toxicity inhibition, namely shaking table oscillation reaction is started after 24h standing period, at the moment, the concentration of the strain starts to be obviously increased, but the strain grows relatively slowly compared with example 3; after two days of growth, the overall OD600The value increased from 0.25 to 0.45, indicating that the strain was not due to the previous S2-The activity of the vaccine is lost, and the activity of the vaccine is gradually restored after a relatively long adaptation period, especially the vaccine contains 300mg/L of S2-The strain concentration of the sample is still steadily increasing.
As can be seen from the example, the strain GXNYJ-DL-1 has strong S resistance2-Toxic capacity, which has now been shown to be able to tolerate S2-The concentration reaches 300 mg/L.
Example 5
Long-chain dicarboxylic acid fermentation sewage treatment by utilizing high-salt-tolerant bacteria GXNYJ-DL-1
The process flow chart of the treatment of the dibasic acid fermentation wastewater is shown in figure 6: the water quality of the wastewater produced by the binary acid fermentation process is as follows: COD 10100mg/L, sulfate 24000mg/L, total salt content 37000mg/L, pH 3.5 and wastewater flow 10 t/h. Adding NaOH into a regulating tank until the pH value is 6.8, flowing into a first-stage aerobic unit, wherein the unit adopts a biological contact oxidation tank process, the strain is high-efficiency halotolerant bacteria GXNYJ-DL-1, the dissolved oxygen is controlled to be more than 2mg/L, the retention time of the wastewater is 72h, the COD of the final effluent is 950mg/L, and flowing into an anaerobic unit. Controlling the dissolved oxygen of the anaerobic unit to be below 0.2mg/L, keeping the wastewater for 90h, keeping the temperature at 30 ℃, and carrying out stripping by adopting methane gas, wherein the volume ratio of the methane stripping gas to the wastewater is 6:1, so that hydrogen sulfide generated by anaerobic reaction is separated from the water phase, enters a gas phase along with methane, and enters a tail gas treatment section. The effluent of the anaerobic unit enters a secondary aerobic unit, the COD of the influent water is 558mg/L, the sulfate content is 2500mg/L, the unit adopts a conventional activated sludge method, conventional biological flora is used, the retention time of the wastewater is 24 hours, the COD of the final effluent is 150mg/L, and the effluent enters a subsequent advanced treatment section.
Advanced oxidation in the advanced treatment section adopts ozone catalytic oxidation, effluent is further treated by BAF, COD of the final effluent is less than or equal to 60mg/L, and total phosphorus is about 60 mg/L. Two-stage chemical phosphorus removal is adopted for the post-phosphorus removal, the chemical agents are ferric chloride and calcium hydroxide conforming agents, the dosage of the first-stage phosphorus removal ferric chloride is 300mg/L, the dosage of the calcium hydroxide is 120mg/L, the dosage of the second-stage phosphorus removal ferric chloride is 100mg/L, the dosage of the calcium hydroxide is 40mg/L, the generated phosphate precipitate is filtered and recovered, COD (chemical oxygen demand) of post-phosphorus removal effluent is less than or equal to 60mg/L, sulfate is about 1700mg/L, the total salt amount is 4200 mg/L, and the total phosphorus is less than or equal to 0.5mg/L, so that the standard discharge is realized.
And the excess sludge generated by the primary aerobic unit, the secondary aerobic unit and the BAF unit is conveyed to a sludge anaerobic oxidation unit by a pump except for normal backflow, and is converted into methane gas by sludge anaerobic oxidation to be recycled to a methane storage tank.
And absorbing tail gas generated by anaerobic treatment by using a lean amine liquid N-methyldiethanolamine solution to obtain methane with the purity of 97%, storing the methane in a methane tank, recycling part of the methane, and using part of the methane as a product. After the solvent absorption section is saturated, the rich amine solution enters a solvent regeneration tower for regeneration, the regenerated lean amine solution returns to the solvent absorption section, the escaped gas is hydrogen sulfide with the purity of about 93 percent, the hydrogen sulfide enters a WSA acid making process, and finally, sulfuric acid with the concentration of about 30 percent is prepared and is recycled as a raw material of a dibasic acid process.
The embodiment shows that the method can effectively treat the long-chain dicarboxylic acid fermentation sewage, and realizes resource recycling while the sewage is discharged up to the standard.
Example 6
High-salt-tolerant bacteria GXNYJ-DL-1 is utilized to treat high-sulfate high-organic wastewater
A certain strand of high sulfate and high organic wastewater is treated by adopting high salt-tolerant bacteria GXNYJ-DL-1, and the sewage treatment flow chart is shown in figure 6.
The water quality of the high-sulfate high-organic wastewater is as follows: 12000mg/L of COD, 35000mg/L of sulfate, 54000mg/L of total salt, 6 of pH and 10t/h of wastewater flow. Adding NaOH into a regulating tank to pH 7, flowing into a first-stage aerobic unit, wherein the unit adopts a biological contact oxidation tank process, the strain is high-efficiency halotolerant bacteria GXNYJ-DL-1, the dissolved oxygen is controlled to be more than 2mg/L, the retention time of the wastewater is 80h, the COD of the final effluent is 980mg/L, and the final effluent flows into an anaerobic unit. Controlling the dissolved oxygen of the anaerobic unit to be below 0.2mg/L, keeping the wastewater for 140h, keeping the temperature at 30 ℃, and carrying out stripping by adopting methane gas, wherein the volume ratio of the methane stripping gas to the wastewater is 7:1, so that hydrogen sulfide generated by anaerobic reaction is separated from the water phase, enters a gas phase along with methane, and enters a tail gas treatment section. The effluent of the anaerobic unit enters a secondary aerobic unit, the COD of the influent water is 718mg/L, the sulfate content is 2650mg/L, the unit adopts a conventional activated sludge method, conventional biological flora is used, the retention time of the wastewater is 24h, the COD of the final effluent is 180mg/L, and the effluent enters a subsequent advanced treatment section.
Advanced oxidation in the advanced treatment section adopts ozone catalytic oxidation, effluent is further treated by BAF, COD of the final effluent is less than or equal to 60mg/L, and total phosphorus is about 5 mg/L. The post-phosphorus removal adopts first-level chemical phosphorus removal, the chemical agents are ferric chloride and calcium hydroxide conforming agents, the dosage of the ferric chloride is 35mg/L, the dosage of the calcium hydroxide is 15mg/L, COD (chemical oxygen demand) of the post-phosphorus removal effluent is less than or equal to 60mg/L, sulfate is about 1750mg/L, the total salt content is 5045 mg/L, and total phosphorus is less than or equal to 0.5mg/L, so that the standard discharge is realized.
And the excess sludge generated by the primary aerobic unit, the secondary aerobic unit and the BAF unit is conveyed to a sludge anaerobic oxidation unit by a pump except for normal backflow, and is converted into methane gas by sludge anaerobic oxidation to be recycled to a methane storage tank.
And absorbing tail gas generated by anaerobic treatment by lean amine liquid N-methyldiethanolamine solution to obtain methane with the purity of 95%, storing the methane in a methane tank, recycling part of the methane, and using part of the methane as a product. After the solvent absorption section absorbs and is saturated, the rich amine solution enters a solvent regeneration tower for regeneration, the regenerated lean amine solution returns to the solvent absorption section, the escaped gas is hydrogen sulfide with the purity of about 94%, the hydrogen sulfide enters a WSA acid preparation process, and finally sulfuric acid with the concentration of about 31% is prepared and is recycled as a dibasic acid process raw material.
Sequence listing
<110> China petrochemical Co., Ltd
China Petroleum & Chemical Corporation Dalian Petrochemical Research Institute
<120> high-salt-tolerance bacterium and application thereof
<130> New patent application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1386
<212> DNA
<213> Halomonas nigrificans
<400> 1
tcgagcggta acaggggtag cttgctaccc gctgacgagc ggcggacggg tgagtaatgc 60
ataggaatct gcccggtagt gggggataac ctggggaaac ccaggctaat accgcatacg 120
tcctacggga gaaagggggc ttcggctccc gctattggat gagcctatgt cggattagct 180
agttggtgag gtaatggctc accaaggcga cgatccgtag ctggtctgag aggatgatca 240
gccacatcgg gactgagaca cggcccgaac tcctacggga ggcagcagtg gggaatattg 300
gacaatgggc gcaagcctga tccagccatg ccgcgtgtgt gaagaaggcc ttcgggttgt 360
aaagcacttt cagcgaggaa gaacgcctag tggttaatac ccattaggaa agacatcact 420
cgcagaagaa gcaccggcta actccgtgcc agcagccgcg gtaatacgga gggtgcgagc 480
gttaatcgga attactgggc gtaaagcgcg cgtaggtggc ttgataagcc ggttgtgaaa 540
gccccgggct caacctggga acggcatccg gaactgtcaa gctagagtgc aggagaggaa 600
ggtagaattc ccggtgtagc ggtgaaatgc gtagagatcg ggaggaatac cagtggcgaa 660
ggcggccttc tggactgaca ctgacactga ggtgcgaaag cgtgggtagc aaacaggatt 720
agataccctg gtagtccacg ccgtaaacga tgtcgaccag ccgttgggtg cctagcgcac 780
tttgtggcga agttaacgcg ataagtcgac cgcctgggga gtacggccgc aaggttaaaa 840
ctcaaatgaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa 900
cgcgaagaac cttacctact cttgacatct acagaagccg gaagagattc tggtgtgcct 960
tcgggaactg taagacaggt gctgcatggc tgtcgtcagc tcgtgttgtg aaatgttggg 1020
ttaagtcccg taacgagcgc aacccttgtc cttatttgcc agcgcgtaat ggcgggaact 1080
ctaaggagac tgccggtgac aaaccggagg aaggtgggga cgacgtcaag tcatcatggc 1140
ccttacgagt agggctacac acgtgctaca atggccggta caaagggttg cgagctcgcg 1200
agagtcagct aatcccgaaa agccggtctc agtccggatc ggagtctgca actcgactcc 1260
gtgaagtcgg aatcgctagt aatcgtgaat cagaatgtca cggtgaatac gttcccgggc 1320
cttgtacaca ccgcccgtca caccatggga gtggactgca ccagaagtgg ttagcctaac 1380
gcaaga 1386