CN116004485A - Pseudomonas, microbial inoculum and method and device for treating dye wastewater by using same - Google Patents

Abstract

The invention discloses a Pseudomonas stutzeri, a microbial inoculum and a method and a device for treating dye wastewater by using the same. The strain is classified as Pseudomonas pseudomonadPseudomonas laurentiana) The strain is named GH-1 and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.26023 and the preservation date of 2022, 11 months and 2 days. The Pseudomonas rourensis of the invention can efficiently degrade naphthalene sulfonic acid organic matters such as 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid, 5-amino naphthalene-2-sulfonic acid and the like in dye intermediate wastewater, thereby solving the problem that the wastewater emission in dye industry is difficult to reach the emission standardIs a problem of (a). The method for treating naphthalene sulfonic acid-containing wastewater by using the strain has the advantages of high tolerance load, good treatment effect, stable effluent quality and the like.

Description

Pseudomonas, microbial inoculum and method and device for treating dye wastewater by using same

Technical Field

The invention belongs to the field of environmental microorganisms, and particularly relates to a method and a treatment device for treating dye wastewater by using Pseudomonas stutzeri and a microbial inoculum.

Background

Along with the accelerated development of the industrialization process, the water pollution is increasingly serious. The textile printing industry is the largest source of pollution and water resource consumer. About 6 tens of thousands of dyes are reported to be discharged into the environment as waste every year worldwide. Every time China produces 1 t dye, 744m of dye is discharged approximately ³ And (5) waste water. The dye released into the water body during the production and use process is about 10% -20%. Meanwhile, since dye molecules generally have stable aromatic structures, degradation is difficult in natural environments, and many dye intermediates may cause cancer, deformity, and mutation to occur, thereby endangering human health. Therefore, the dye wastewater becomes one of important factors for threatening the safety of water environment in China.

Naphthalene sulfonic acid organic matter is naphthalene dye and dye intermediate widely used in dye industry, and is naphthalene nucleus with naphthalene ring as mother nucleus and containing one or several sulfonic acid groups (-SO) ₃ H) The aromatic ring may also contain other substituents such as amino, hydroxyl and the like, is a raw material for synthesizing chemicals such as cationic dyes, azo dyes, acid mediator dyes and the like, and can also be used as an organic intermediate of phenols, leather tanning agents, pesticides, dyes and the like. The industrial wastewater has the characteristics of complex components, high chromaticity, high water solubility, strong polarity, poor biodegradability, high toxicity, carcinogenesis and the like; structurally, the presence of sulfonic acid groups renders such compounds highly mobile in water, while the stable structure of the naphthalene ring and carbon-sulfur bonds results in recalcitrance, which means that naphthalene sulfonic acid compounds can accumulate in the environment, which would pose a serious threat to the environment and human health if not reasonably remedied.

There are many methods for treating dye intermediate wastewater at present, but the following methods are mainly used in consideration of efficiency and cost: physical, chemical and biological methods. The physical method is mainly used for pretreatment of dye wastewater, is generally simple to operate and low in treatment cost, but has insufficient depth for treating the dye wastewater and poor removal effect. The chemical method is to generate a dye wastewater treatment unit based on chemical reaction by adding chemical reagents into dye wastewater, and convert pollutants into harmless substances. Advanced oxidation technology is widely used for removing organic matters in wastewater at present. However, in the actual water treatment process, the degradation cost is high by adopting the advanced oxidation technology alone, and only partial oxidation of the organic pollutants is completed instead of complete mineralization. A series of intermediate oxidation products accumulate during advanced oxidation and further advanced treatments are still necessary. The method for treating the wastewater by utilizing the metabolism of microorganisms has the advantages of low cost, good treatment effect, simple operation and management and the like. The strain capable of degrading naphthalene sulfonic acid organic matters is screened and becomes a research hotspot for domestic and foreign scholars.

Disclosure of Invention

The invention aims to: aiming at the defects of the prior art, the Lawrence river mouth pseudomonas for degrading naphthalene sulfonic acid compounds in dye intermediate wastewater is provided, and a novel technology is provided for treating the dye intermediate wastewater by adopting a biological method.

Specifically, the application provides Pseudomonas pseudomonad belonging to the genus Pseudomonas, which is named as Pseudomonas pseudomonad(Pseudomonas laurentiana) The strain is named GH-1 and is preserved in China general microbiological culture Collection center (CGMCC), the preservation number is CGMCC No.26023, and the preservation date is 2022, 11 and 02 days. The GH-1 is obtained by screening activated sludge in an aeration tank of a sewage treatment system of a dye factory. The strain is determined to be the Pseudomonas stutzeri by morphological characteristics, physiological and biochemical characteristics and 16S rDNA sequence determination and phylogenetic analysis.

The strain GH-1 has the following physical and chemical characteristics: the bacterial stain is gram negative, short rod-shaped under the lens, no spore, single or short chain arrangement (shown in figure 1); the strain forms milky or light yellow colony with raised center, tidy peripheral edges, smooth surface and opaque surface on agar culture medium.

The invention provides the Pseudomonas pseudomonad for the Larens river mouthPseudomonas laurentiana) Naphthalene sulfonic acid organic matters in dye intermediate wastewater can be used as a sole carbon source for growth; when the initial concentration of naphthalene sulfonic acid organic matters is 0.5-1 g/L, laurens Heteropap Pseudomonas(Pseudomonas laurentiana) The degradation of naphthalene sulfonic acid organic matters can reach more than 93% after 48 hours.

The invention further provides a microbial inoculum prepared from the Pseudomonas stutzeri.

The specific method comprises the following steps:

(1) Selecting Pseudomonas rourensis GH-1 from LB solid medium, shake culturing in LB liquid medium to logarithmic phase to obtain bacterial liquid;

(2) Transferring the bacterial liquid obtained in the step (1) into a seed tank for culture according to the transfer volume ratio of 5% -10%;

(3) Inoculating the bacterial liquid cultured in the seed tank into a fermentation tank for expansion culture, wherein the transfer volume ratio is 5% -10%, and the bacterial agent can be obtained after fermentation is completed.

Wherein, the LB liquid medium comprises the following components: 10g/L peptone, 5g/L yeast extract and 10g/L NaCl; adding agar with the volume ratio of 2% into LB liquid medium to obtain LB solid medium, and shake culturing under the following conditions: the culture pH value is 7.0-7.5, and the culture temperature is 30-35 ℃.

The culture medium components of the seed tank and the fermentation tank are the same, and the culture medium components are as follows: glucose 8g/L, yeast extract 5g/L, naCl g/L, K ₂ HPO ₄ 1g/L、CaCO ₃ 2g/L、MgSO ₄ 0.2g/L, and pH value is 7.0-7.5; the temperature of the seed tank is controlled to be 25-35 ℃, the stirring speed is 180-250 rpm, the dissolved oxygen DO is controlled to be 3-4 mg/L, and the culture time is 24-48 h; in the fermentation tank, the fermentation conditions are as follows: stirring at 180-240 rpm, culturing at 25-35 ℃, controlling dissolved oxygen DO at 4-6 mg/L, and fermenting for 48-96 h; number of cells after fermentationUp to 10 ⁹ And (3) the concentration of the active component is more than one/mL.

The invention also provides application of the Laurens estuary pseudomonas or the microbial inoculum in degrading naphthalene sulfonic acid compounds in dye intermediate wastewater.

Specifically, the naphthalene sulfonic acid compound is any one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid.

During treatment, preparing the Pseudomonas stutzeri into seed liquid, and adding the seed liquid into the wastewater; or adding the microbial inoculum into the wastewater; wherein, the OD600 of the prepared seed liquid is 1.0-1.5, and the inoculation amount is 5-10%; when the wastewater is treated, the temperature is 20-35 ℃ and the pH is 7-8.

The invention also provides a dye wastewater treatment device, which comprises an MBR reactor, wherein the MBR reactor is provided with a microbial inoculum adding device, and the microbial inoculum adding device is provided with the Pseudomonas roursonii or the microbial inoculum prepared by the Pseudomonas roursonii.

The first stage: introducing naphthalene sulfonic acid compounds with the concentration of 50-200 mg/L into an MBR biological pond, regulating the pH value of wastewater to 7-8, adding strains or microbial agents with the volume ratio of 5% -10% into the MBR reactor, culturing and enriching microorganisms, enabling the strains or microbial agents to be attached to the surface of a filler to form a biological film, and collecting and discharging qualified wastewater;

step two, gradually increasing the concentration of naphthalene sulfonic acid compounds in the MBR to 200-1000 mg/L, so that the quality of effluent water is gradually stabilized, and collecting and discharging qualified wastewater;

thirdly, introducing wastewater to be treated into the MBR, and collecting and discharging qualified wastewater;

wherein, MBR pond operating parameter is: 2-4 mg/L of dissolved oxygen, running temperature of 20-35 ℃ and HRT of 24-48 h;

the temperature of the wastewater to be treated is 15-40 ℃; the pH value of the wastewater is 4-9; the concentration of pollutants to be degraded in the wastewater to be treated is 0.2-1.0g/L; the naphthalene sulfonic acid compound is one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid; the wastewater to be treated is wastewater containing any one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid.

The beneficial effects are that: the invention relates to a Pseudomonas pseudomonad at the mouth of Lorentz riverPseudomonas laurentiana) The GH-1 has stronger adaptability and tolerance to living environment, can degrade naphthalene sulfonic acid organic compounds of wastewater under extreme acid-base environment conditions, solves the problems of strong inhibition effect and low degradation efficiency of pH on functional microorganisms, and provides efficient microorganism germplasm resources for degradation of organic matters in industrial wastewater; meanwhile, the degradation efficiency of the Laurens estuary pseudomonas GH-1 and the microbial inoculum on the 1, 5-naphthalene disulfonic acid, the 1, 6-naphthalene disulfonic acid, the 2-amino-1-naphthalene sulfonic acid and the 5-amino naphthalene-2-sulfonic acid can reach more than 93 percent within 48 hours, and the invention solves the problem that the waste water emission in dye industry is difficult to reach the standard and is difficult to discharge, and has important significance in the aspects of protecting ecological environment, protecting the health of people, promoting the harmonious symbiosis of people and nature and the like.

Drawings

The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.

FIG. 1 is a colony morphology of Pseudomonas stutzeri GH-1;

FIG. 2 is a phylogenetic tree of Pseudomonas stutzeri GH-1;

FIG. 3 is a graph showing the effect of Pseudomonas stutzeri GH-1 on degrading naphthalene sulfonic acid organic pollutants;

FIG. 4 is the effect of different pH on the degradation effect of Pseudomonas stutzeri GH-1;

FIG. 5 is the effect of this different temperature on the degradation effect of Pseudomonas stutzeri GH-1;

FIG. 6 is a graph showing the effect of Pseudomonas stutzeri on the degradation of actual dye intermediate wastewater pollutants;

FIG. 7 is a graph showing the effect of Pseudomonas stutzeri on contaminant degradation during MBR reactor startup;

FIG. 8 is a graph showing the effect of Pseudomonas larensis on contaminant degradation in an MBR reactor.

Description of the embodiments

The following examples facilitate a better understanding of the present invention, but are not intended to limit the same. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the examples described below, unless otherwise specified, were purchased from conventional biochemical reagent stores. The quantitative tests in the following examples were all set up in triplicate and the results averaged.

Example 1 isolation, screening and identification of degradation strains.

(1) Screening, separating and purifying degradation bacteria

10mL of activated sludge in an aeration tank of a wastewater treatment system of a dye factory is taken and placed in 90 mL inorganic salt liquid medium containing 1, 5-naphthalene disulfonic acid (400 mg/L), and the mixture is subjected to shaking enrichment culture for 7d by a constant temperature shaking table at 30 ℃ and 180 r/min. Preparing an inorganic salt culture medium, controlling the concentration of 1, 5-naphthalene disulfonic acid in the culture medium to be 200mg/L, 500mg/L, 750mg/L, 900mg/L and 1000mg/L in sequence, inoculating 5% of inoculum size into the inorganic salt liquid culture medium according to the volume ratio, and continuously enriching and culturing at 30 ℃ and 180 rpm. And (3) after a proper amount of enriched bacterial liquid is subjected to gradient dilution, the bacterial liquid is coated on a flat plate of an inorganic salt solid culture medium containing 1000mg/L1, 5-naphthalene disulfonic acid, the culture is carried out at a constant temperature of 30 ℃, after bacterial colonies grow out, streak separation is carried out, single bacterial colonies are picked up and connected to an LB solid culture medium, the pure culture of the bacterial strain is obtained through flat plate streak separation and purification, and the pure culture is named GH-1 and is stored in a refrigerator at a temperature of 4 ℃ for standby.

Wherein, the inorganic salt liquid culture medium comprises the following components: (NH) ₄ ) ₂ SO ₄ 1.0g/L、CaCl ₂ •2H ₂ O 0.04g/L、FeSO ₄ •7H ₂ O 0.005g/L、Na ₂ HPO ₄ •12H ₂ O 1.0g/L、MgSO ₄ •7H ₂ O0.8 g/L and KH ₂ PO ₄ 2.0g/L, pH 7.0, 2% agar was added to the solid medium, and the medium was sterilized at 121℃for 30 minutes.

LB solid medium: 10g/L peptone, 5g/L yeast extract and 10g/L NaCl, pH 7.0, 2% agar was added, and the medium was sterilized at 121℃for 30 minutes.

(2) Identification of degradation strains

After sequencing, the gene sequences of the strains were analyzed by comparison of the blowing program in the GenBank database with the nucleic acid data in NCBI. The DNA sequences determined using MEGA6 software were systematically developed and homologously analyzed against the 16SrDNA gene sequence-related species representative strains in the standard database, by selecting appropriate sequences. And confidence level detection is carried out, and the bootstrap data set is 1000 times. The phylogenetic tree of the strains was drawn (see FIG. 2). The strain GH-1 has the same branch with the standard strain MG719526.1, and the similarity reaches more than 99 percent, so the strain GH-1 is identified as the Pseudomonas pseudomonad of Lorentia river mouthPseudomonas laurentiana). The strain is preserved in 2022.11.02 in China general microbiological culture Collection center (CGMCC) with a preservation number of 26023, and the preservation unit address is 1 st Cello 3 in the Korean area of Beijing, and the postal code is 100101.

Bacterial colony morphology of the strain GH-1 on the LB plate is light yellow, round, smooth in surface and neat in edge, and the bacterial colony is moist and glossy (shown in figure 1). The main biological characteristics are: the terminal flagellum, the bacillus-free, short rod-shaped bacteria, gram-negative, can grow with the dye organic intermediate as the sole carbon source.

Example 2 degradation performance study of Pseudomonas stutzeri GH-1 on target pollutants.

Inoculating Pseudomonas pseudomonad GH-1 at Lorentia river mouth into LB culture medium, shake culturing at 30deg.C under 180rpm, centrifuging at 8000rpm for 15 min until the thallus enters late stage of logarithmic phase, skimming supernatant, suspending the thallus in sterile inorganic salt liquid culture medium by vortex vibration method, centrifuging repeatedly, washing for three times, suspending the thallus in sterile inorganic salt liquid culture medium again (regulating the amount of culture medium, and controlling the OD600 of the bacterial suspension to be about 1.0), and obtaining seed liquid.

The inorganic salt culture medium is prepared, the initial concentration of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid in the culture medium is respectively 1000mg/L, 1000mg/L and 500mg/L, and the pH value of the culture medium is adjusted to 7. Inoculating 5% of the inoculum size into each culture medium, shake culturing at 30deg.C and 180rpm, sampling at fixed time, and detecting degradation condition, wherein the degradation effect is shown in figure 3.

As can be seen from FIG. 3, the strain GH-1 can effectively degrade 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid, and the degradation rates after 48 hours are 94.5%, 93.1%, 96.3% and 95.9%, respectively.

This example illustrates the isolation of Pseudomonas stutzeriPseudomonas laurentiana) GH-1 can utilize naphthalene sulfonic acid organic matters as the sole carbon source for growth and reproduction, and effectively degrade naphthalene sulfonic acid organic matters in dye intermediate wastewater.

Example 3 pH effect on degradation effect performance study.

Strains GH-1 of Pseudomonas stutzeri cultivated in LB medium to logarithmic phase were inoculated into inorganic salt medium containing naphthalene sulfonic acid organic matters (1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid, 5-amino naphthalene-2-sulfonic acid) in an inoculum size of 5% by volume, the initial concentration of each naphthalene sulfonic acid organic matter in the medium was 500mg/L, pH values were adjusted to 4, 5, 6, 7, 8, 9 by 1mol/L hydrochloric acid and sodium hydroxide, shaking culture was carried out at 30℃and 180rpm, and the concentration of each naphthalene sulfonic acid organic matter was sampled and measured and degradation rate was calculated after 48 hours, as shown in FIG. 4.

As can be seen from FIG. 4, under the conditions that the initial pH is 7 and 8, the degradation rate of the bacterial strain GH-1 on various pollutants is over 95 percent; when the initial pH is 5, 6 and 9, the degradation rate of the bacterial strain GH-1 on various pollutants is more than 85%, and the bacterial strain GH-1 still has higher degradation efficiency; however, when the pH was lowered to 5 or less, the strain degradation effect was rapidly lowered to 50% or less.

This example demonstrates that the proper pH range for the Pseudomonas stutzeri GH-1 to degrade naphthalene sulfonic acid organics is 5-9, and the removal efficiency is highest in a neutral environment, and the removal capacity of naphthalene sulfonic acid organics is significantly reduced in an acidic environment.

Example 4 study of the effect of temperature on degradation effect.

Strains GH-1 of Pseudomonas stutzeri cultivated in LB medium to logarithmic phase were inoculated into inorganic salt medium containing naphthalene sulfonic acid organic matters (1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid) in an inoculum size of 5% by volume, the initial concentration of each naphthalene sulfonic acid organic matter in the medium was 500mg/L, each naphthalene sulfonic acid organic matter was cultivated in shaking tables at 15 ℃, 20 ℃, 25 ℃,30 ℃, 35 ℃, 40 ℃ and 180r/min, and after 48 hours, the concentration of each naphthalene sulfonic acid organic matter was measured by sampling and the degradation rate was calculated as shown in FIG. 5.

As shown in FIG. 5, the degradation rate of the strain GH-1 on each pollutant gradually increases when the temperature is within the range of 15-30 ℃, and the degradation rate of each pollutant is highest and is about 95% when the temperature is 30 ℃; when the temperature is within the range of 30-40 ℃, the degradation rate of the bacterial strain GH-1 on various pollutants is gradually reduced.

This example shows that the optimum temperature for the degradation of 1, 5-naphthalenedisulfonic acid, 1, 6-naphthalenedisulfonic acid, 2-amino-1-naphthalenesulfonic acid, 5-aminonaphthalene-2-sulfonic acid by Pseudomonas aeruginosa GH-1 is 30 ℃.

Example 5 preparation of degrading bacterial agent.

Selecting Pseudomonas stutzeri GH-1 from LB solid medium, shake culturing at 30deg.C and 180rpm to logarithmic phase; transferring GH-1 bacterial liquid in logarithmic growth phase with transfer volume ratio of 10% into a seed tank for culturing, controlling the temperature of the seed tank to 30deg.C, stirring at 220rpm, and controlling dissolved oxygen DO at 4mg/L. Inoculating the bacterial liquid cultured in the seed tank into a fermentation tank for expansion culture in an inoculum size of 5%, and controlling fermentation conditions: stirring speed is 220rpm, culture temperature is 30 ℃, and dissolved oxygen DO is 6mg/L; the effective viable count in the fermentation liquor after the fermentation is finished is 10 ⁹ And (3) packaging the fermentation broth after the fermentation broth is taken out of the tank to obtain the degradation microbial inoculum.

Wherein, the culture medium components of the seed tank and the fermentation tank are the same, and the culture medium components are: glucose 8g/L, yeast extract 5g/L, naCl g/L, K ₂ HPO ₄ 1g/L、CaCO ₃ 2g/L、MgSO ₄ 0.2g/L, and pH value is 7.0-7.5.

Example 6 application effect study of strain GH-1 in actual wastewater treatment.

The source of the treated water is sewage treatment plant inlet water containing 1, 5-naphthalene disulfonic acid (1.5 g/L), 1, 6-naphthalene disulfonic acid (1.1 g/L), 2-amino-1-naphthalene sulfonic acid (0.8 g/L) and 5-amino naphthalene-2-sulfonic acid (0.6 g/L), and the pH of the waste water is about 7-8; the high-efficiency degrading bacterium GH-1 bacterial liquid obtained in example 4 was inoculated into waste water according to an inoculum size of 10% by volume, the control group was inoculated with activated sludge, and subjected to shaking culture at 30℃and 180rpm, after 48 hours, the concentration of the contaminant was measured and the degradation rate was calculated, and the result was shown in FIG. 6.

As shown in FIG. 6, the degradation effect of the conventional sludge on pollutants in wastewater is not obvious, the removal rate is only about 25%, and the removal rates of the pollutants in wastewater to which the GH-1 bacterial liquid of the invention is added are all above 90%, and the removal rates of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid are 93.2%, 92.4%, 94.5% and 91.0%, respectively. Therefore, the microbial inoculum produced by the strain GH-1 can be applied to the treatment of dye intermediate wastewater of naphthalene sulfonic acid organic compounds.

Example 7 degradation effects of strain GH-1 in MBR reactor were studied.

This example shows an operation for treating dye intermediate wastewater, particularly naphthalene sulfonic acid wastewater, with Pseudomonas lazii in an MBR reactor.

The method adopts simulated wastewater, and the water quality is as follows: 200mg/L, sodium chloride 5g/L, NH ₄ SO ₄ 1.0 g/L，KH ₂ PO ₄ 0.5 g/L，MgSO ₄ •7H ₂ O0.2 g/L. Adjusting the pH value of the wastewater to 7-8 by using 1mol/L hydrochloric acid and sodium hydroxide, introducing the wastewater into an MBR reactor with the treatment capacity of 5L, adding a GH-1 microbial agent into the reactor, wherein the volume percentage of the GH-1 microbial agent in the wastewater is 5%, culturing and enriching microorganisms, enabling the strains to be attached to the surface of a filler to form a biological film, enriching for 48-72 h, and tracking the degradation condition of 1, 5-naphthalene disulfonic acid. And after the degradation rate of 1, 5-naphthalene disulfonic acid is stabilized to be more than 60%, gradually increasing the concentration of 1, 5-naphthalene disulfonic acid in the wastewater to be 500mg/L, 750mg/L, 900mg/L and 1000mg/L in sequence, tracking the degradation condition, as shown in figure 7, after the reactor is operated stably, respectively introducing the catalyst containing naphthalene sulfonic acid organic matters (1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid,2-amino-1-naphthalene sulfonic acid and 5-aminonaphthalene-2-sulfonic acid), the concentration of each naphthalene sulfonic acid organic matter is 500mg/L, and the degradation condition is tracked after 48 hours, as shown in figure 8. Wherein, MBR pond operating parameter: 2-4 mg/L of dissolved oxygen, running temperature of 20-35 ℃ and HRT of 24-48 h.

As can be seen from FIGS. 7 and 8, the GH-1 microbial inoculum of the invention is added into the MBR reactor, and after the reactor runs stably, the GH-1 microbial inoculum can efficiently degrade pollutants in wastewater, and the removal rates of 1, 5-naphthalenedisulfonic acid, 1, 6-naphthalenedisulfonic acid, 2-amino-1-naphthalenesulfonic acid and 5-aminonaphthalene-2-sulfonic acid are 97.4%, 97.8%, 98.9% and 98.2%, respectively. The experimental data show that the GH-1 microbial inoculum has good prospect in degrading naphthalene sulfonic acid dye intermediate wastewater in an MBR reactor.

The invention provides a thought and a method for degrading naphthalene sulfonic acid compounds in dye intermediate wastewater, and particularly provides a method and a plurality of ways for realizing the technical scheme, the method and the method are only preferred embodiments of the invention, and it should be pointed out that a plurality of improvements and modifications can be made by one of ordinary skill in the art without departing from the principle of the invention, and the improvements and modifications are also considered as the protection scope of the invention. The components not explicitly described in this embodiment can be implemented by using the prior art.

Claims (10)

1. Pseudomonas pseudomonad strain, named as Pseudomonas pseudomonad strainPseudomonas laurentiana) The strain is named GH-1 and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.26023 and the preservation date of 2022, 11 months and 2 days.

2. A microbial agent comprising pseudomonas stutzeri according to claim 1.

3. The method for preparing the microbial inoculum of claim 2, which is characterized by comprising the following steps:

(1) Selecting Pseudomonas rourensis GH-1 from LB solid medium, shake culturing in LB liquid medium to logarithmic phase to obtain bacterial liquid;

(2) Transferring the bacterial liquid obtained in the step (1) into a seed tank for culture according to the transfer volume ratio of 5% -10%;

(3) Inoculating the bacterial liquid cultured in the seed tank into a fermentation tank according to the transfer volume ratio of 5% -10% for expansion culture, and obtaining the bacterial agent after fermentation.

4. The method for preparing the microbial inoculum according to claim 3, wherein the components of the LB liquid medium are as follows: 10g/L peptone, 5g/L yeast extract and 10g/L NaCl; adding agar with the volume ratio of 2% into LB liquid medium to obtain LB solid medium; the culture conditions of shaking culture are as follows: the culture pH value is 7.0-7.5, and the culture temperature is 30-35 ℃.

5. The method for producing a microbial inoculum according to claim 3, wherein the culture medium components of the seed tank and the fermentation tank are the same, and the culture medium components are: glucose 8g/L, yeast extract 5g/L, naCl g/L, K ₂ HPO ₄ 1g/L、CaCO ₃ 2g/L、MgSO ₄ 0.2g/L, and pH value is 7.0-7.5; the temperature of the seed tank is controlled to be 25-35 ℃, the stirring speed is 180-250 rpm, the dissolved oxygen DO is controlled to be 3-4 mg/L, and the culture time is 24-48 h; in the fermentation tank, the fermentation conditions are as follows: stirring at 180-240 rpm, culturing at 25-35 ℃, controlling dissolved oxygen DO at 4-6 mg/L, and fermenting for 48-96 h; after fermentation, the number of the thalli reaches 10 ⁹ And (3) the concentration of the active component is more than one/mL.

6. Use of the pseudomonas larensis of claim 1 or the microbial inoculum of claim 2 for degrading naphthalene sulfonic acid compounds in dye intermediate wastewater.

7. The use according to claim 6, wherein the naphthalene sulfonic acid compound is any one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid, 5-aminonaphthalene-2-sulfonic acid.

8. A method for degrading naphthalene sulfonic acid compounds in dye intermediate wastewater, which is characterized in that the microbial inoculum of claim 2 is added into the wastewater according to an inoculation amount of 5-10% by volume; when the wastewater is treated, the temperature is 20-35 ℃ and the pH is 7-8.

9. Dye wastewater treatment device, characterized in that the device comprises an MBR reactor, wherein the MBR reactor is provided with a microbial inoculum adding device, and the microbial inoculum adding device is provided with the Pseudomonas roursonii according to claim 1 or the microbial inoculum according to claim 2.

10. A method for degrading naphthalene sulfonic acid compounds in dye intermediate wastewater by using the device of claim 9, which is characterized by comprising the following steps:

the first stage: introducing 50-200 mg/L naphthalene sulfonic acid compound into an MBR biological pond, regulating the pH value of wastewater to 7-8, adding 5-10% by volume of the strain of claim 1 or the microbial inoculum of claim 2 into the MBR reactor, culturing and enriching microorganisms, enabling the strain or microbial inoculum to be attached to the surface of a filler to form a biological film, and collecting and discharging qualified wastewater;

step two, gradually increasing the concentration of naphthalene sulfonic acid compounds in the MBR to 200-1000 mg/L, so that the quality of effluent water is gradually stabilized, and collecting and discharging qualified wastewater;

thirdly, introducing wastewater to be treated into the MBR, and collecting and discharging qualified wastewater;

wherein, MBR pond operating parameter is: 2-4 mg/L of dissolved oxygen, running temperature of 20-35 ℃ and HRT of 24-48 h;

the temperature of the wastewater to be treated is 15-40 ℃; the pH value of the wastewater is 4-9; the concentration of pollutants to be degraded in the wastewater to be treated is 0.2-1.0g/L; the naphthalene sulfonic acid compound is one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid; the wastewater to be treated is wastewater containing any one or more of 1, 5-naphthalene disulfonic acid, 1, 6-naphthalene disulfonic acid, 2-amino-1-naphthalene sulfonic acid and 5-amino naphthalene-2-sulfonic acid.

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