CN113462622A - Pseudomonas for efficiently degrading various aromatic pollutants and application thereof - Google Patents
Pseudomonas for efficiently degrading various aromatic pollutants and application thereof Download PDFInfo
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- 230000000593 degrading effect Effects 0.000 title claims abstract description 24
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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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- C02F2101/30—Organic compounds
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- C02F2101/00—Nature of the contaminant
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Abstract
The invention provides pseudomonas for efficiently degrading various aromatic pollutants and application thereof. The invention discloses pseudomonasPseudomonas gessardii The preservation number of AD1 is CCTCC NO: m2021841. The pseudomonas isPseudomonas gessardiiThe AD1 has strong environmental adaptability, and can efficiently degrade various aromatic pollutants such as toluene, p-hydroxybenzoic acid and the like at the temperature of 25-35 ℃ and the pH value of 6.0-8.0; when the temperature is 30 ℃ and the pH value is 7.0, the degradation rate of different aromatic pollutants can reach 80-90 percent, the toxicity of sewage can be reduced, and the method can be used for alkali treatment of lignocelluloseThe degradation of aromatic pollutants in the treatment of various industrial wastewater such as black liquor, coking wastewater and the like has higher application value.
Description
Technical Field
The invention belongs to the technical field of environmental microorganisms, and particularly relates to pseudomonas for efficiently degrading various aromatic pollutants and application thereof.
Background
Aromatic compounds are a general name of a class of compounds containing a benzene ring structure, and not only widely exist in natural environment, but also widely applied to human social industry, agricultural production and daily life. Aromatic compounds can be roughly classified into the following three groups according to their structures: simple aromatic compounds, polycyclic aromatic compounds, and heterocyclic compounds. Due to the stability of the benzene ring structure, part of aromatic compounds can remain in the environment for a long time, and great threat is formed to the ecological environment and human health. For example, there may be several threats to human health, such as neurotoxicity, blood toxicity, reproductive toxicity and carcinogenicity, and efficient treatment techniques and processes are needed.
The treatment method of the aromatic pollutants mainly comprises a physical method, a chemical method and a biological method. Among them, the physical and chemical methods include adsorption, coagulation, catalytic oxidation, etc., and the biological method is mainly performed by assimilation and metabolism of aerobic and anaerobic bacteria. The aromatic pollutants are various in variety, and microorganisms have great difference on the metabolism mode of different aromatic pollutants, so that different treatment effects are presented. Compared with physical degradation methods and chemical degradation methods, biological degradation methods have the characteristics of high efficiency, low cost, no secondary pollution, good adaptability to various environments and the like, and gradually become one of the most promising environment restoration technologies.
The treatment of aromatic pollutants by biodegradation depends heavily on the separation and screening of high-efficiency degrading strains. Currently, aromatic pollutant degrading strains widely reported mainly include pseudomonas, bacillus, rhodococcus, sphingolipid and the like, wherein the pseudomonas is a key research object in related fields due to multiple functional strain types, remarkable degrading effect and wide environmental adaptability, and also appears in more related patent applications. However, in consideration of the diversity and difficult degradability of aromatic pollutants, the current biological treatment process still needs to develop more broad-spectrum efficient degradation strains and process flows. The growth and function of microorganisms are often influenced by various environmental factors, such as temperature, pH, substrate type, etc., and in practice, it is common for multiple contaminants to be present simultaneously, possibly in a synergistic or antagonistic manner. Therefore, it is an important subject to be solved to fully utilize the microbial resources in the nature, to develop new high-efficiency degradation bacterial strains of aromatic pollutants, and to clarify the degradation functional characteristics and environmental adaptability.
Disclosure of Invention
In view of the above problems, the present invention aims to provide a pseudomonas strain capable of efficiently degrading various aromatic pollutants and an application thereof, wherein the pseudomonas strain is used for effectively degrading various simple aromatic compounds in industrial wastewater. .
The invention also aims to provide application of the strain in treatment of industrial wastewater containing aromatic pollutants.
In order to realize the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention provides a bacterial strain AD1 for efficiently degrading aromatic pollutants, which is classified into the following strains by 16S rDNA sequence alignment and combining morphological characteristics, physiological and biochemical characteristics of the bacterial strainPseudomonas gessardii. The strain is preserved in China Center for Type Culture Collection (CCTCC) with the strain name ofPseudomonas gessardii AD1, date of deposit: 2021, 7/8, accession no: CCTCC NO: m2021841.
The pseudomonas isPseudomonas gessardii AD1, characterized in that it is a gram-negative bacterium. The colony morphology is white circular, the surface is smooth, the edge is neat, and the colony size is about 2.0-4.0 mm. The cells were observed under a microscope to be in the form of short rods with sizes of (0.4 to 1.0) × (1.0 to 3.5) μm.
The pseudomonas isPseudomonas gessardii The growth temperature of the AD1 is 10-45 ℃, and the optimal growth temperature is 25-35 ℃.
The pseudomonas isPseudomonas gessardii AD1, the temperature range for degrading aromatic compounds is 10-45 ℃, preferably 25-30 ℃.
The pseudomonas isPseudomonas gessardii AD1, the growth pH value is 4.0-10.0, preferably 6.0-8.0.
The pseudomonas isPseudomonas gessardii The AD1 has 80-90% degradation rate for different aromatic compounds at 30 deg.C and pH of 7.0.
In a second aspect, the invention provides the pseudomonasPseudomonas gessardii Use of AD1 in the treatment of industrial waste water containing a single aromatic contaminant and industrial waste water containing multiple aromatic contaminants.
The application comprises the following steps:
(1) culturing pseudomonas AD1 with cane molasses culture medium to obtain microbial agent;
(2) and (3) inoculating the microbial agent into the industrial wastewater to be treated by using water according to a proportion (5-10%) to degrade the target pollutants.
The concentration of the cane molasses in the cane molasses culture medium is 20.0-50.0 g/L.
Pseudomonas in the microbial agentPseudomonas gessardiiThe content of AD1 is not less than 5 × 108CFU/mL。
Compared with the prior art, the pseudomonas for efficiently degrading various aromatic pollutants and the application thereof have the following beneficial effects:
(1) various common aromatic pollutants in the industrial wastewater are efficiently degraded, the degradation rate can reach 85-90%, and the application value is high;
(2) high treatment efficiency, good economic benefit, convenient operation and no pollution.
Drawings
FIG. 1 is the environmental suitability for growth of Pseudomonas AD1 in example 2. A is different temperature and B is different pH value.
FIG. 2 shows the environmental suitability of Pseudomonas AD1 for degrading single aromatic compounds (p-hydroxybenzoic acid as an example) in example 2. A is different temperature and B is different pH value.
FIG. 3 is the morphology of the growing colonies of Pseudomonas AD1 on the plate in example 1.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the accompanying drawings and specific embodiments.
Example 1: screening, performance verification and identification of pseudomonas AD1
The media and components used were as follows:
enriching a liquid culture medium: 5.5 g/L glucose, 10.0 g/L (NH)4)2SO4，0.25 g/L KH2PO4，1.0 g/L K2HPO411.5 g/L sodium succinate, 0.2 g/L MgCl21.0 mL/L iron solution and 1.0 mL/L trace element solution, and the pH value is 7.0. Preparing an iron solution: 4 mL/L37% HCl and 5.0 g/L FeSO4·7H2And O. Composition of the trace element solution: MnSO4·4H2O 100.0 mg/L，ZnCl2 70.0 mg/L，Na2MoO4·2H2O 35.0 mg/L，CuSO4·5H2O 30.0 mg/L，CoCl2·6H2O 200.0 mg/L，H3BO3 60.0 mg/L，NiCl2·6H2O25.0 mg/L, HCl 0.9 mL/L at 37% concentration.
Separating and purifying a solid culture medium: 15.0 g/L agar powder, 2.5 g/L (NH)4)2SO4，0.25 g/L KH2PO4，1.0 g/L K2HPO45.5 g/L glucose, 11.5 g/L sodium succinate, 0.2 g/L MgCl21.0 mL/L iron solution, 1.0 mL/L trace element solution, pH 7.0, the compositions of the iron solution and the trace element solution were the same as above.
Sugar cane molasses liquid culture medium: 20.0 to 50.0 g/L cane molasses and 0.75 g/L KH2PO4，3.0 g/L K2HPO4，0.2 g/L MgCl21.0 mL/L iron solution, 1.0 mL/L trace element solution, pH value7.0, the composition of the iron solution and the trace element solution was the same as described above.
Verifying the liquid culture medium: hydroxybenzoic acids of different concentrations (0.5 g/L, 1 g/L, 2.5 g/L and 5 g/L), 0.25 g/L KH2PO4，1.0 g/L K2HPO45.5 g/L glucose, 11.5 g/L sodium succinate, 0.2 g/L MgCl21.0 mL/L iron solution, 1.0 mL/L trace element solution, pH 7.0, the compositions of the iron solution and the trace element solution were the same as above.
The prepared culture medium is sterilized by high pressure steam at 115 deg.C for 20 min.
Enrichment culture: the screening sample is collected in petrochemical engineering wastewater, and 5 mL of the collected sample is inoculated into a triangular flask filled with 95 mL of enrichment liquid culture medium for culture. The culture conditions are 30 ℃, the rotating speed is 200 r/min, and the culture is finished after 48 hours. And then inoculating 5 ml of culture into a fresh enrichment culture medium for passage, wherein the passage condition is consistent with the culture condition, and finishing the culture after passage for 2-3 times. The final culture is used for the subsequent isolation of useful microorganisms.
Separation and purification: and (3) performing gradient dilution on the culture subjected to enrichment culture, and uniformly coating the culture on a solid culture medium plate. After coating, the plate is inverted in a constant temperature incubator at 30 ℃, colonies with different shapes and sizes are picked according to the colony morphology on the plate after culturing for 48 hours, and the colonies are marked, purified and numbered for preservation. The 4 strains were co-isolated and named AD1, AD2, AD3 and AD4, respectively.
And (3) performance measurement: the 4 screened strains are respectively cultured by a cane molasses liquid culture medium for 12 hours, and then are respectively inoculated into a reaction bottle filled with 200 mL of verification liquid culture medium according to the inoculation amount of 5 percent, and the reaction temperature is controlled at 30 ℃. Parahydroxybenzoic acid is taken as an example for verifying the degradation performance of the strain on aromatic contaminants. It was confirmed that different concentrations of parahydroxybenzoic acid (0.5 g/L, 1 g/L, 2.5 g/L and 5 g/L) were added to the medium, samples were taken after 24 hours of culture, the concentration of the remaining parahydroxybenzoic acid was determined by liquid chromatography, and the degradation rate was calculated. The degradation rates of p-hydroxybenzoic acid by the 4 strains obtained by screening are shown in Table 1. The AD1 has the most outstanding degradation capability, and the concentration range of the hydroxybenzoic acid is 0.5-2.5 g/L, and the degradation capability is higher than 85%. The p-hydroxybenzoic acid has a certain inhibiting effect on 4 strains of bacteria, and the degradation rate of the p-hydroxybenzoic acid is gradually reduced along with the increase of the initial concentration. When the initial concentration is higher than 5 g/L, the degradation effect is not significant.
TABLE 14 Properties of the strain for the degradation of para-hydroxybenzoic acid
Identification of strain AD 1: the genome of the strain is extracted, and the taxonomical characteristics of the strain AD1 are identified by a 16S sequencing method. Genome extraction the 16S rDNA fragment of strain AD1 was amplified using primers 27f (agatttgattctcggctca) and 1492r (ggttactctgttacgactt) using the bacterial genome extraction kit from AXYGEN. The total volume of the PCR reaction system was 30. mu.L, and contained 1. mu.L of template DNA, 15. mu.L of Primer mix, 1. mu.L of each of the upstream and downstream primers, and ddH2O to 12. mu.L. PCR procedure: circulating for 30 times at 98 deg.C for 5 min, 98 deg.C for 30 s, 55 deg.C for 30 s, and 68 deg.C for 90 s, at 68 deg.C for 10 min, and at 16 deg.C for 10 min. Purification and sequencing of PCR products were performed by Shanghai Biotech, Inc. The 16S rDNA length 1248 bp of the strain AD1 is obtained by sequencing, and the nucleic acid sequence is shown as SEQID NO. 1. Through the analysis of Nucleotide BLAST, the base consistency of the sequence and pseudomonas CIP 105469 (NCBI serial number: NR-024928.1) in NCBI database is 1240 bp, and the similarity reaches 99%. The AD1 strain which is determined and screened by combining the morphological characteristics and the growth characteristics of the bacteria is pseudomonasPseudomonasgessardiiIs named asPseudomonasgessardii AD1。
Pseudomonas spPseudomonasgessardiiThe biological characteristics of AD1 are as follows: gram-negative, short rod-shaped, the size of thallus is (0.4-1.0) × (1.0-3.5) mum, the diameter of colony is 2.0-4.0 mm, the colony is round, the surface is smooth, and the edge is neat.
Pseudomonas screened outPseudomonasgessardiiAD1, depository: china Center for Type Culture Collection (CCTCC) address: storage of the eight Wuhan university in Wuchang district, Wuhan city, Hubei provinceDate: 2021, 7/8, accession no: CCTCC NO: m2021841.
Example 2: environmental suitability and function of pseudomonas AD1
In order to verify the environmental suitability of the pseudomonas AD1, the growth and the aromatic pollutant degradation performance (taking p-hydroxybenzoic acid as an example) of a strain of the pseudomonas AD1 are respectively examined under the conditions of different temperatures and different pH values. The specific experiment is as follows:
the environmental adaptability of the growth of the pseudomonas AD 1.
(1) Temperature: pseudomonas AD1 was inoculated in the enrichment medium, pH was controlled to 7.0, and culture temperatures were 10 deg.C, 15 deg.C, 20 deg.C, 25 deg.C, 30 deg.C, 35 deg.C, 40 deg.C, and 45 deg.C, respectively. A certain amount of sample was taken out every 3 hours to measure the growth of the cells, and the culture was terminated after 24 hours, and the results are shown in FIG. 1 (A). The pseudomonas AD1 can grow at 10-45 ℃, well grows within the range of 25-40 ℃, and OD is obtained after culturing for 24 h600The temperature reaches 2.87-4.35, wherein the optimal growth temperature is 35 ℃.
(2) pH value: inoculating Pseudomonas AD1 in enrichment culture medium, controlling culture temperature at 30 deg.C and pH at 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0, respectively. A certain amount of samples were taken every 3 hours to determine the growth of the strain, and the culture was terminated at 24 hours, and the results are shown in FIG. 1 (B). The pseudomonas AD1 can grow in the pH value range of 4.0-10.0, and can grow well in the pH value range of 6.0-8.0, and the OD is obtained after culturing for 24 h600The pH value of the growth medium reaches 2.91-4.07, wherein the optimum growth pH value is 7.0.
And secondly, the pseudomonas AD1 has environmental suitability for degrading p-hydroxybenzoic acid.
(1) Temperature: preparing a verification culture medium with the volume of 200 mL and the pH value of 7.0 in a reaction bottle, and adding 1 g/L p-hydroxybenzoic acid; the temperature of the reaction flask is controlled at 10 ℃, 15 ℃, 20 ℃, 25 ℃, 30 ℃, 35 ℃, 40 ℃ and 45 ℃ respectively, the pseudomonas AD1 with the same volume is inoculated, and the degradation reaction of the p-hydroxybenzoic acid is started. Degradation Process samples were taken every 6 h to determine the concentration of p-hydroxybenzoic acid remaining in the system, and the results are shown in FIG. 2 (A). The pseudomonas AD1 can degrade p-hydroxybenzoic acid at 10-45 ℃, the degradation effect is good at 25-35 ℃, the degradation rate reaches 91.45-96.37%, and the optimal degradation temperature is 30 ℃.
(2) pH value: similarly, a reaction bottle is provided with a verification culture medium with the volume of 200 mL, and p-hydroxybenzoic acid with the concentration of 1 g/L is added; the reaction temperature was controlled at 30 ℃ and the pH of the medium in the reaction flask was adjusted to 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0, respectively, and the same volume of Pseudomonas AD1 was inoculated to initiate the degradation reaction of p-hydroxybenzoic acid. Degradation Process samples were taken every 6 h to determine the concentration of p-hydroxybenzoic acid remaining in the system, and the results are shown in FIG. 2 (B). The pseudomonas AD1 can degrade p-hydroxybenzoic acid within the pH value range of 4.0-10.0, the degradation effect is good within the pH value range of 6.5-8.5, the degradation rate reaches 90.03-94.42%, and the optimal degradation pH value is 7.0.
Example 3: performance of Pseudomonas AD1 in degrading different aromatic pollutants
Toluene, m-diphenol and vanillic acid are respectively selected, and the degradation performance of pseudomonas AD1 on single aromatic pollutants is examined.
Firstly, the performance of the pseudomonas AD1 in degrading toluene. Culturing Pseudomonas AD1 to OD with cane molasses medium600The value is more than 5.0, and the obtained culture is a microbial agent. 200 mL of validation medium was placed in the reaction flask and toluene (0.05 g/L, 0.1 g/L, 0.5 g/L, and 1 g/L) was added at various concentrations. Inoculating microbial agent at an inoculation ratio of 5%, culturing for 24 h, determining the concentration of residual toluene by high performance liquid chromatography, and calculating the degradation rate. After 24 hours of treatment, the degradation rate of toluene with different concentrations exceeds 85 percent (except 1 g/L). The initial concentration of 1 g/L, the degradation rate of pseudomonas AD1 to toluene is reduced to 45.6%.
And secondly, the pseudomonas AD1 has the performance of degrading m-diphenol. The pseudomonas AD1 is cultured by a cane molasses culture medium to obtain the microbial inoculum. 200 mL of validation medium was placed in the reaction flask and resorcinol 3 (0.1 g/L, 0.5 g/L, 1 g/L and 2.5 g/L) was added at various concentrations. Inoculating microbial agent at an inoculation ratio of 5%, culturing for 24 h, determining the concentration of residual resorcinol by high performance liquid chromatography, and calculating the degradation rate. After 24 hours of treatment, the degradation rate of resorcinol at different concentrations exceeds 90% (except 2.5 g/L). At an initial concentration of 2.5 g/L, the degradation rate of the pseudomonas AD1 to the resorcinol is reduced to 58.3%.
And thirdly, the pseudomonas AD1 is used for degrading vanillic acid. The pseudomonas AD1 is cultured by a cane molasses culture medium to obtain the microbial inoculum. 200 mL of validation medium was prepared in a reaction flask, and different concentrations of vanillic acid (0.1 g/L, 0.5 g/L, 1 g/L, and 2.5 g/L) were added. Inoculating microbial agent at an inoculation ratio of 5%, culturing for 24 h, determining the concentration of the residual vanillic acid by means of high performance liquid chromatography, and calculating the degradation rate. After 24 hours of treatment, the degradation rate of vanillic acid with different concentrations exceeds 90% (except 2.5 g/L). The degradation rate of the pseudomonad AD1 to the vanillic acid is reduced to 77.4% when the initial concentration is 2.5 g/L.
Example 4: application of pseudomonas AD1 in treatment of wastewater containing multiple aromatic pollutants
The application of the pseudomonas AD1 in treating black liquor generated by alkaline pretreatment of lignocellulose. The pseudomonas AD1 is cultured by a cane molasses culture medium to obtain the microbial inoculum. The black liquor produced by alkaline pretreatment of 2L of lignocellulose is respectively filled in 2 100L sealed plastic barrels. The experimental group is inoculated with the microbial inoculum according to the inoculation ratio of 5 percent, and the control group is added with sterile water according to the same ratio. And after 24 hours, respectively measuring the residual concentrations of the aromatic pollutants in the barrels of the experimental group and the control group by using liquid chromatography, and calculating the degradation rate. After 24 hours of treatment, the average degradation rate of aromatic pollutants in an experimental group reaches 87.9%, which proves the effectiveness of pseudomonas AD1 in degrading the aromatic pollutants in the alkaline lignocellulose pretreatment black liquor, and synchronously treats various aromatic compounds, wherein the degradation rate is higher than that of a single compound, and a synergistic degradation effect exists.
And secondly, the pseudomonas AD1 is applied to the treatment of coking wastewater. The pseudomonas AD1 is cultured by a cane molasses culture medium to obtain the microbial inoculum. 2L of coking wastewater is respectively filled into 2 100L sealed plastic barrels. The experimental group was inoculated with a microbial inoculum at an inoculation ratio of 5%, and the control group was added with sterile water at the same ratio. And after 24 hours, measuring the residual concentrations of different aromatic pollutants in the barrels of the experimental group and the control group by using liquid chromatography, and calculating the degradation rate. After 24 hours of treatment, the average degradation rate of aromatic pollutants in an experimental group reaches 83.4%, the effectiveness of pseudomonas AD1 in degrading aromatic pollutants in coking wastewater is proved, various aromatic compounds are synchronously treated, and the degradation rate of the aromatic compounds is higher than that of a single compound.
Sequence listing
<110> Yuhuang ecological environmental science and technology Limited in Foshan City
<120> pseudomonas for efficiently degrading various aromatic pollutants and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1248
<212> DNA
<213> Pseudomonas gessardii AD1
<400> 1
gatcctggct cagattgaac gctggcggca ggcctaacac atgcaagtcg agcggtagag 60
agaagcttgc ttctcttgag agcggcggac gggtgagtaa tgcctaggaa tctgcctggt 120
agtgggggat aacgttcgga aacggacgct aataccgcat acgtcctacg ggagagagca 180
ggggaccttc gggccttgcg ctatcagatg agcctaggtc ggattagcta gttggtgggg 240
taatggctca ccaaggcgac gatccgtaac tggtctgaga ggatgatcag tcacactgga 300
actgagagac ggtccagact cctacgggag gcagcagtgg ggaatattgg acaatgggcg 360
aaagcctgat ccagccatgc cgcgtgtgtg aagaaggtct tcggattgta aagcacttta 420
agttgggagg aagggttgta gattaatact ctgcaatttt gacgttactg acagaataag 480
caccggctaa ctctgtgcca gcagccgcgg taatacggag ggtgcaagcg ttaatcggaa 540
ttactgggcg taaagcgcgc gtaggtggtt agttacgttg gatgtgaaat ccccgggctc 600
aacctgggaa ctgcattcaa aactgactga ctagagtatg gtagagggtg gtggaatttc 660
ctgtgtagcg gtgaaatgcg tagatatagg aaggaacacc agtggcgaag gcgaccacct 720
ggactgatac tgacactgcg gtgcgaaagc gtggggagca aacaggatta gataccctgg 780
tagtccacgc cgtaaacgat gtcaactagc cgttgggagc cttgagctct tagtggcgca 840
gctaacgcat taagttgacc gcctggggag tacggccgca aggttagaac tcaaatgaat 900
tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc 960
ttaccaggcc ttgacatcca atgaactttc tagagataga ttggtgcctt cgggaacatt 1020
gagacaggtg ctgcatggct gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgt 1080
aacgagcgca acccttgtcc ttagttacca gcacgttatg gtgggcactc taaggagact 1140
gccggtgaca aaccggcgga aggtggggat gacgtcaagt catcatggcc cttacggcct 1200
gggctacaca cgtgctacaa tggtcggtac agagggttgc caagccgc 1248
Claims (9)
1. Pseudomonas for degrading aromatic pollutantsPseudomonas gessardiiAD1, preserved in China Center for Type Culture Collection (CCTCC), and the preservation date is as follows: 2021, 7/8, accession no: CCTCC NO: m2021841.
2. A microbial agent comprising the Pseudomonas bacterium according to claim 1Pseudomonas gessardii AD1。
3. The microbial inoculant according to claim 2, wherein pseudomonads are present in said microbial inoculantPseudomonas gessardiiThe content of AD1 is not less than 5 × 108 CFU/mL。
4. The Pseudomonas bacterium according to any one of claims 1 to 2Pseudomonas gessardii Use of AD1, characterized in that said pseudomonasPseudomonas gessardii AD1 is used to degrade aromatic contaminants.
5. The Pseudomonas bacterium according to any one of claims 1 to 2Pseudomonas gessardii Use of AD1, characterized in that said pseudomonasPseudomonas gessardii AD1 is used in the treatment of industrial wastewater containing one or more aromatic contaminants.
6. The use of claim 5, wherein said Pseudomonas spPseudomonas gessardii AD1 also includes one or more of the following features:
(1) the growth temperature is 10-45 ℃;
(2) the growth pH value ranges from 4.0 to 10.0;
(3) the temperature range for degrading the aromatic pollutants is 10-45 ℃;
(4) the pH value range of the aromatic pollutants is 4.0-10.0.
7. The use of claim 6, wherein said Pseudomonas spPseudomonas gessardii The AD1 has the degradation rate of 85-90% for different aromatic pollutants under the conditions of 30 ℃ and pH value of 7.0.
8. Use according to claim 7, characterized in that it comprises the following steps:
(1) culturing pseudomonas AD1 with cane molasses culture medium to obtain microbial agent;
(2) proportionally inoculating the microbial inoculum to the industrial wastewater to be treated by using water, and degrading the target pollutants.
9. The use of claim 8, further comprising one or more of the following features:
(1) the concentration of the cane molasses in the cane molasses culture medium is 20.0-50.0 g/L;
(2) the inoculation proportion of the microbial inoculum is 5-10% of the volume of the industrial wastewater to be treated.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114410552A (en) * | 2022-03-28 | 2022-04-29 | 佛山市玉凰生态环境科技有限公司 | Pseudomonas gainsteri for oxidizing sulfur and application of pseudomonas gainsteri in sulfur reduction in black and odorous water body air |
| CN115197864A (en) * | 2021-12-31 | 2022-10-18 | 广州理格致生物科技有限公司 | Pseudomonas and method for degrading monocyclic aromatic hydrocarbon by using same |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1729286A (en) * | 2002-02-08 | 2006-02-01 | 南邦环境与生物技术株式会社 | Pseudomonas sp. FK916 producing decomposition enzyme of organic matter and method for decomposition of organic waste using the same |
| CN101629154A (en) * | 2009-08-03 | 2010-01-20 | 新疆农业科学院微生物应用研究所 | Psychrotrophs and application thereof in wastewater treatment |
| US20130280228A1 (en) * | 2009-04-24 | 2013-10-24 | Dupont Nutrition Biosciences Aps | Feed supplement |
| CN112322551A (en) * | 2020-11-25 | 2021-02-05 | 南京农业大学 | Bacterium with multiple degradation effects on undesirable components of tobacco leaves and application |
-
2021
- 2021-09-03 CN CN202111029374.2A patent/CN113462622B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1729286A (en) * | 2002-02-08 | 2006-02-01 | 南邦环境与生物技术株式会社 | Pseudomonas sp. FK916 producing decomposition enzyme of organic matter and method for decomposition of organic waste using the same |
| US20130280228A1 (en) * | 2009-04-24 | 2013-10-24 | Dupont Nutrition Biosciences Aps | Feed supplement |
| CN101629154A (en) * | 2009-08-03 | 2010-01-20 | 新疆农业科学院微生物应用研究所 | Psychrotrophs and application thereof in wastewater treatment |
| CN112322551A (en) * | 2020-11-25 | 2021-02-05 | 南京农业大学 | Bacterium with multiple degradation effects on undesirable components of tobacco leaves and application |
Non-Patent Citations (3)
| Title |
|---|
| HAIYING HUANG ET AL.: "A novel Pseudomonas gessardii strain LZ-E simultaneously degrades naphthalene and reduces hexavalent chromium", 《BIORESOURCE TECHNOLOGY》 * |
| JIRI WALD ET AL.: "Pseudomonads Rule Degradation of Polyaromatic Hydrocarbons in Aerated Sediment", 《FRONTIERS IN MICROBIOLOGY》 * |
| 杨峰山等: "两株苯唑草酮降解细菌的筛选与降解能力测定", 《生物学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115197864A (en) * | 2021-12-31 | 2022-10-18 | 广州理格致生物科技有限公司 | Pseudomonas and method for degrading monocyclic aromatic hydrocarbon by using same |
| CN115197864B (en) * | 2021-12-31 | 2023-12-19 | 广州畅培优食品研究有限公司 | Pseudomonas and method for degrading monocyclic aromatic hydrocarbon by using pseudomonas |
| CN114410552A (en) * | 2022-03-28 | 2022-04-29 | 佛山市玉凰生态环境科技有限公司 | Pseudomonas gainsteri for oxidizing sulfur and application of pseudomonas gainsteri in sulfur reduction in black and odorous water body air |
| CN114410552B (en) * | 2022-03-28 | 2022-06-17 | 佛山市玉凰生态环境科技有限公司 | Pseudomonas gainsteri for oxidizing sulfur and application of pseudomonas gainsteri in sulfur reduction in black and odorous water body air |
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