Synthetic flora and application thereof in degradation of sulfamethoxazole in sewage

Abstract

The invention provides a synthetic flora and application thereof in degradation of sulfamethoxazole in sewage, and belongs to the technical field of sewage treatment. According to the present invention, at least one bacterium selected from the group consisting of Nocardiopsis corallicola, Gordonia hydrophila and Halioglobus pacificus and Dokdonia fugitiva are adopted to construct the synthetic bacterial flora, such that the effective degradation of the sulfamethoxazole in the sewage can be achieved so as to prevent the persistent pollution of the sulfamethoxazole in the sewage to the environment.

Description

Technical Field

[0001] This invention relates to the field of wastewater treatment technology, and in particular to a synthetic microbial community and its application in the degradation of sulfamethoxazole in wastewater. Background Technology

[0002] Sulfamethoxazole (SMX), a sulfonamide antibiotic, is commonly used to treat and prevent Pneumocystis pneumonia, and is mainly discharged into sewage through medical channels.

[0003] However, during the wastewater treatment process, due to the stable molecular structure of sulfamethoxazole, existing wastewater treatment methods cannot effectively degrade sulfamethoxazole, resulting in persistent environmental pollution from sulfamethoxazole discharged into wastewater. Summary of the Invention

[0004] This invention proposes a synthetic microbial community and its application in the degradation of sulfamethoxazole in wastewater, employing... Nocardiopsis coralliicola , Gordoniahydrophobica , Peaceful haloglobus At least one of the bacteria and Dokdonella fugitive Constructing synthetic microbial communities can effectively degrade sulfamethoxazole in wastewater, thereby preventing persistent environmental pollution from sulfamethoxazole in wastewater.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: A synthetic microbial community, characterized in that it comprises a first degrading functional bacterium and a second degrading functional bacterium, wherein the first degrading functional bacterium includes... Nocardiopsis coralliicola , Gordoniahydrophobica , Peaceful haloglobus At least one of the bacteria, the second degrading bacteria include Dokdonella fugitive .

[0006] Preferably, the ratio of viable bacteria of the first degrading functional bacteria to the second degrading functional bacteria in the above-mentioned synthetic microbial community is 0.2~5:1.

[0007] Further preferred, the first degrading functional bacteria include Nocardiopsis coralliicola , Gordoniahydrophobica , Peaceful haloglobus When two types of bacteria are present, the live bacteria ratio of the two types of bacteria is 0.01 to 100:1.

[0008] Further preferred, the first degrading functional bacteria include Nocardiopsis coralliicola , Gordoniahydrophobica as well as Peaceful haloglobus hour, Nocardiopsis coralliicola , Gordoniahydrophobica as well as Peaceful haloglobus The live bacteria ratio is 0.01~100:0.01~100:1.

[0009] The present invention also provides the application of the above-mentioned synthetic microbial community in the degradation of sulfamethoxazole in wastewater.

[0010] In one implementation of the above application, the synthetic microbial community is inoculated into the wastewater of a wastewater treatment system to degrade sulfamethoxazole in the wastewater. Preferably, the inoculation concentration of the synthetic microbial community is 10. 6 ~10 8 cells / mL.

[0011] It should be noted that the wastewater treatment system inoculated with the synthetic microbial community provided by this invention is not only applicable to sequencing batch reactors, but also to mainstream wastewater treatment processes such as continuous flow activated sludge systems, membrane bioreactors, biofilters, or moving bed biofilm reactors, and does not rely on electrochemical or physicochemical enhancement methods. The inoculation form of the synthetic microbial community can be liquid culture, lyophilized powder, or immobilized carrier. As long as the effective colonization and functional expression of the synthetic microbial community in the wastewater treatment system are ultimately achieved, they all belong to the equivalent embodiments of this invention.

[0012] Compared with the prior art, the present invention has the following beneficial effects.

[0013] The present invention provides a synthetic microbial community, in which the first degrading functional bacteria include Nocardiopsis coral reef , Gordoniahydrophobica as well as Peaceful haloglobus All of them can convert it into the intermediate metabolite 1,2,4-phenylpyrogallol through their respective initial degrading enzyme systems (i.e., monooxygenase SadA, dioxygenase SadB, and FMN reductase SadC). The secondary degrading bacteria contain... Dokdonella fugitive The invention can further degrade the intermediate metabolite 1,2,4-phenylpyrogallol into β-ketoadipic acid. β-ketoadipic acid, as a natural metabolic intermediate, is environmentally harmless and readily biodegradable. It can be further degraded into succinyl-CoA and acetyl-CoA (intermediate metabolites commonly found in bacteria) by indigenous bacteria in the wastewater treatment system, and then enter the tricarboxylic acid cycle for complete degradation. Therefore, the synthetic bacteria provided by this invention can effectively and completely degrade sulfamethoxazole in wastewater, thereby preventing persistent environmental pollution caused by sulfamethoxazole in wastewater. Attached Figure Description

[0014] Figure 1 This is a schematic diagram of the biodegradation pathway of sulfamethoxazole provided in the embodiments of this application; Figure 2 This is a schematic diagram of the core species of the metabolic complementarity network provided in the embodiments of this application; Figure 3 This is a schematic diagram illustrating the degradation capabilities of strains G, H, N, and D on pollutants provided in the embodiments of this application; Figure 4 This is a schematic diagram of MICOM simulation of 7 combinations of sulfamethoxazole uptake flux provided in the embodiments of this application; Figure 5 This is a schematic diagram showing the change in sulfamethoxazole concentration in the pure culture system provided in the embodiments of this application; Figure 6 This is a schematic diagram of the concentration change of sulfamethoxazole in a sequencing batch reactor provided in the embodiments of this application. Detailed Implementation

[0015] In the specification and claims of this invention, the terms "first" and "second," etc., are used to distinguish different objects, rather than to describe a specific order of objects.

[0016] In the embodiments of this application, "and / or" indicates a relationship between objects. For example, A and / or B can represent the following three situations: A exists alone, B exists alone, and A and B exist simultaneously.

[0017] In the embodiments of this application, the terms "exemplary" or "for example" are used to indicate that something is an example, illustration, or description. Any embodiment or design that is described as "exemplary" or "for example" in the embodiments of this application should not be construed as being more preferred or advantageous than other embodiments or design. Specifically, the use of the terms "exemplary" or "for example" is intended to present the relevant concepts in a specific manner.

[0018] Example 1: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Gordoniahydrophobica The second degrading bacteria include Dokdonella fugitive The ratio of viable bacteria with the first to the second degrading function in the synthetic microbial community was 1:1.

[0019] in, Gordoniahydrophobica Purchased from the China Industrial Microbial Culture Collection Center, with accession number CICC 24170; Dokdonella fugitive Purchased from Beijing Bio-Bio Biotechnology Co., Ltd.

[0020] Example 2: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Peaceful haloglobus The second degrading bacteria include Dokdonella fugitive The ratio of viable bacteria with the first to the second degrading function in the synthetic microbial community was 1:4.

[0021] in, Peaceful haloglobus and Dokdonella fugitive All were purchased from Beijing Biowell Biotechnology Co., Ltd.

[0022] Example 3: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Nocardiopsis coralliicola The second degrading bacteria include Dokdonella fugitive The ratio of viable bacteria with the first to the second degrading function in the synthetic microbial community was 1:5.

[0023] in, Nocardiopsis coralliicola and Dokdonella fugitive All were purchased from Beijing Biowell Biotechnology Co., Ltd.

[0024] Example 4: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Gordoniahydrophobica and Peaceful haloglobus The second degrading bacteria include Dokdonella fugitive The ratio of viable primary degrading bacteria to secondary degrading bacteria in the synthetic microbial community was 1:1; among the primary degrading bacteria... Gordoniahydrophobica and Peaceful haloglobus The live bacteria ratio is 1:1.

[0025] The above Gordoniahydrophobica , Peaceful haloglobus as well as Dokdonella fugitive For the purchase channels, please refer to the relevant descriptions in Examples 1-3 above.

[0026] Example 5: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Gordoniahydrophobica and Nocardiopsis coralliicola The second degrading bacteria include Dokdonella fugitive The viable ratio of the first-degrading functional bacteria to the second-degrading functional bacteria in the synthetic microbial community was 5:1. Among the first-degrading functional bacteria... Gordoniahydrophobica and Nocardiopsis coralliicola The live bacteria ratio is 1:1.

[0027] The above Gordoniahydrophobica , Nocardiopsis coralliicola as well as Dokdonella fugitive For the purchase channels, please refer to the relevant descriptions in Examples 1-3 above.

[0028] Example 6: This example describes a synthetic microbial community, including a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Peaceful haloglobus and Nocardiopsis coralliicola The second degrading bacteria include Dokdonella fugitive The viable ratio of the first-degrading functional bacteria to the second-degrading functional bacteria in the synthetic microbial community was 3:1. Among the first-degrading functional bacteria... Peaceful haloglobus and Nocardiopsis coral reef The live bacteria ratio is 1:1.

[0029] The above Peaceful haloglobus , Nocardiopsis coralliicola as well as Dokdonella fugitive For the purchase channels, please refer to the relevant descriptions in Examples 1-3 above.

[0030] Example 7: This example describes a synthetic bacterial community, which includes a first degrading functional bacterium and a second degrading functional bacterium. The first degrading functional bacterium includes... Gordoniahydrophobica , Peaceful haloglobus as well as Nocardiopsis coralliicola The second degrading bacteria include Dokdonella fugitive The ratio of viable primary degrading bacteria to secondary degrading bacteria in the synthetic microbial community was 1:1. Among the primary degrading bacteria... Gordoniahydrophobica , Peaceful haloglobus as well as Nocardiopsis coral reef The ratio of live bacteria is 1:1:1.

[0031] The above Gordoniahydrophobica , Peaceful haloglobus , Nocardiopsis coral reef as well as Dokdonella fugitive For the purchase channels, please refer to the relevant descriptions in Examples 1-3 above.

[0032] Example 8: This example describes a synthetic bacterial community. The difference between this example and Example 4 is that: in the first degrading functional bacteria... Gordoniahydrophobica and Peaceful haloglobus The live bacteria ratio was 0.01:1.

[0033] Example 9: This example describes a synthetic bacterial community. The difference between this example and Example 4 is that: in the first degrading functional bacteria... Gordoniahydrophobica and Peaceful haloglobus The live bacteria ratio is 100:1.

[0034] Example 10: This example describes a synthetic bacterial community. The difference between this example and Example 5 is that: in the first degrading functional bacteria... Gordoniahydrophobica and Nocardiopsis coralliicola The live bacteria ratio was 0.01:1.

[0035] Example 11: This example describes a synthetic bacterial community. The difference between this example and Example 5 is that: in the first degrading functional bacteria... Gordoniahydrophobica and Nocardiopsis coralliicola The live bacteria ratio is 100:1.

[0036] Example 12: This example describes a synthetic bacterial community. The difference between this example and Example 6 is that: in the first degrading functional bacteria... Peaceful haloglobus and Nocardiopsis coralliicola The live bacteria ratio was 0.01:1.

[0037] Example 13: This example describes a synthetic bacterial community. The difference between this example and Example 6 is that: in the first degrading functional bacteria... Peaceful haloglobus and Nocardiopsis coralliicola The live bacteria ratio is 100:1.

[0038] Example 14: This example describes a synthetic bacterial community. The difference between this example and Example 7 is that: in the first degrading functional bacteria... Gordoniahydrophobica , Peaceful haloglobus as well as Nocardiopsis coral reef The live bacteria ratio was 0.01:0.01:1.

[0039] Example 15: This example describes a synthetic bacterial community. The difference between this example and Example 7 is that: in the first degrading functional bacteria... Gordoniahydrophobica , Peaceful haloglobus as well as Nocardiopsis coral reef The live bacteria ratio is 100:100:1.

[0040] Example 16: This example describes the application of a synthetic microbial community described in Example 1 in the degradation of sulfamethoxazole in wastewater.

[0041] Specifically, according to 10 6 ~10 8 An inoculation concentration of cells / m will be used to inoculate synthetic microbial communities into the wastewater treatment system to achieve the degradation of sulfamethoxazole in the wastewater.

[0042] like Figure 2 As shown, during the degradation of sulfamethoxazole in wastewater by the synthetic microbial community, the first degrading bacteria can convert it into the intermediate metabolite 1,2,4-phenylpyrogallol through the initial degrading enzyme system (i.e., monooxygenase SadA, dioxygenase SadB, and FMN reductase SadC). The second degrading bacteria can further degrade the intermediate metabolite 1,2,4-phenylpyrogallol into β-ketoadipic acid. β-ketoadipic acid, as a natural metabolic intermediate, has no environmental hazards and is extremely biodegradable. It can be further degraded into succinyl-CoA and acetyl-CoA (intermediate metabolites commonly found in bacteria), and then enter the tricarboxylic acid cycle to achieve complete degradation.

[0043] It should be noted that the wastewater treatment system inoculated with the synthetic microbial community provided in this embodiment is not only applicable to sequencing batch reactors, but also to mainstream wastewater treatment processes such as continuous flow activated sludge systems, membrane bioreactors, biofilters, or moving bed biofilm reactors. The inoculation form can be liquid culture, lyophilized powder, or immobilized carrier. As long as the effective colonization and functional expression of the synthetic microbial community in the wastewater treatment system are ultimately achieved, it is considered an equivalent implementation of this embodiment.

[0044] Example 17: This example describes the application of a synthetic microbial community as described in Example 2 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0045] Example 18: This example describes the application of a synthetic microbial community as described in Example 3 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is the same as described in Example 8, and will not be repeated here.

[0046] Example 19: This example describes the application of a synthetic microbial community as described in Example 4 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0047] Example 20: This example describes the application of a synthetic microbial community as described in Example 5 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0048] Example 21: This example describes the application of a synthetic microbial community as described in Example 6 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0049] Example 22: This example describes the application of a synthetic microbial community described in Example 7 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0050] Example 23: This example describes the application of a synthetic microbial community described in Example 8 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is the same as described in Example 8, and will not be repeated here.

[0051] Example 24: This example describes the application of a synthetic microbial community as described in Example 9 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is the same as described in Example 8, and will not be repeated here.

[0052] Example 25: This example describes the application of a synthetic microbial community described in Example 10 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0053] Example 26: This example describes the application of a synthetic microbial community described in Example 11 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0054] Example 27: This example describes the application of a synthetic microbial community described in Example 12 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0055] Example 28: This example describes the application of a synthetic microbial community described in Example 13 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is the same as described in Example 8, and will not be repeated here.

[0056] Example 29: This example describes the application of a synthetic microbial community as described in Example 14 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0057] Example 30: This example describes the application of a synthetic microbial community as described in Example 15 in the degradation of sulfamethoxazole in wastewater. The specific application of the synthetic microbial community in this example is similar to that in Example 8, and will not be repeated here.

[0058] Experimental Example: This experimental example describes the four types of bacteria upon which the synthetic microbial communities described in Examples 1-7 above are based. Nocardiopsis coralliicola , Gordoniahydrophobica、Halioglobus pacificus as well as Dokdonella fugitive The screening process was conducted, and the degradation effect of the synthetic microbial community described in Examples 1-7 on sulfamethoxazole (hereinafter referred to as SMX) in wastewater was verified.

[0059] The following are the four types of bacteria mentioned above. Nocardiopsis coralliicola (hereinafter referred to as N) Gordoniahydrophobica (Hereinafter referred to as G) 、Peaceful globe (hereinafter referred to as H) and Dokdonella fugitivaThe screening process (hereinafter referred to as D).

[0060] Step 1: Strain screening based on functional gene criteria.

[0061] Screening for key functional genes in the genome that are associated with the SMX degradation pathway.

[0062] For bacteria with primary degradation function, the selection criteria are that the strain carries the sadABC gene cluster (SMX degradation function gene), which can encode monooxygenase SadA, dioxygenase SadB, and FMN reductase SadC.

[0063] The filtering process corresponding to the above filtering criteria is as follows.

[0064] Global metagenomic sequence data of activated sludge were collected. Metagenome assembled genomes (MAGs) were obtained through sequence assembly, redundancy removal, and binning. High-quality MAGs with genome integrity ≥90% and contamination level <5% were selected for subsequent analysis.

[0065] The aforementioned global activated sludge metagenomic data was collected from the NCBI database (an existing database). The search criteria were: 'activated sludge metagenome' [Organism] AND (metagenome [Filter] AND 'bioproject sra' [Filter]). The raw sequences collected using these search criteria were screened, and sequences with the detection type 'WGS' metagenomics, paired-end sequencing, read length greater than 100 bp, and base pair count greater than 200,000,000 were selected as the aforementioned global activated sludge metagenomic data.

[0066] The protein sequences of the above-mentioned high-quality MAGs were predicted using Prodigal software, and then compared with known SMX degradative enzyme sequences (monooxygenase SadA, dioxygenase SadB, and FMN reductase SadC) using BLASTP software. The sequence similarity was ≥50%, and the expected value was ≤1×10⁻⁶. -15 MAGs containing multiple sequences can degrade SMX into 1,2,4-pyrogallol, thus achieving the first functional degradation function ( Figure 1 The degradation function of ) will be considered. Multiple strains containing the MAGs to which the above sequences belong will be selected as the first preselected strains.

[0067] For bacteria with secondary degradation function, the screening criteria are that the strain carries hydroxyquinone-1,2-dioxygenase and maleic acid reductase.

[0068] The filtering process corresponding to the above filtering criteria is as follows.

[0069] The enzyme numbers corresponding to the above screening criteria in the KEGG database are EC 1.13.11.37 and EC 1.3.1.32, respectively. The screening process is as follows: Functional annotation of the protein sequences of all high-quality MAGs is performed according to the KEGG database. MAGs annotated with both EC 1.13.11.37 and EC 1.3.1.32 are selected as functional bacteria for the second functional module. It should be understood that the aforementioned KEGG database is a commonly used existing database in this technical field, and the specific application process of the aforementioned KEGG database in the experimental examples of this application will not be elaborated upon.

[0070] Specifically, a search of the KEGG database revealed that 1,2,4-phenylpyrogallol can be degraded to maleic acid by hydroxyquinone-1,2-dioxygenase, and maleic acid is then degraded to β-ketoadipic acid by maleic acid reductase. The corresponding enzyme numbers in the KEGG database for these two processes are EC 1.13.11.37 and EC 1.3.1.32, respectively. Next, high-quality MAGs were functionally annotated. If both enzymes were annotated, then the corresponding MAG could perform the conversion of 1,2,4-phenylpyrogallol to β-ketoadipic acid. Therefore, multiple strains containing the aforementioned corresponding MAGs were selected as the second pre-selected strains.

[0071] Step 2: Strain screening based on quorum sensing criteria.

[0072] The screening criteria for strains based on quorum sensing standards are: simultaneously encoding quorum sensing signal molecule synthetic proteins and receiver proteins, and the number of these two types of protein sequences accounts for ≥50% of the total number of protein sequences in MAGs.

[0073] The screening process described above involves collecting synthetic and receiver protein sequences of quorum sensing signaling molecules from the SigMol and Quorumpeps databases. These known protein sequences are used to train machine learning models based on the k-nearest neighbor and random forest algorithms. These trained models are then used to identify protein sequences in MAGs (Mutable Gram-Oriented Groups). Sequences that are simultaneously identified as positive by both models are selected. These sequences are then compared with the AS-QSB and UniProt databases to determine their properties (synthetic or receiver proteins). Multiple MAGs containing both synthetic and receiver protein sequences, with the proportion of these two types of sequences ≥ 50% of the total protein sequence count, are identified as quorum-sensing bacteria.

[0074] It should be noted that the SigMol, Quorumpeps, AS-QSB, and UniProt databases mentioned above are commonly used techniques in this technical field, and the experimental examples in this application will not further elaborate on these databases. The two models mentioned above are existing models, and their sources can be found in the published literature Ying Jin, Wenkang Chen, Jie Hu, Jinfeng Wang, Hongqiang Ren, Constructions of quorumsensing signaling network for activated sludge microbial community, ISMECommunications, Volume 4, Issue 1, January 2024, ycae018, https: / / doi.org / 10.1093 / ismeco / ycae018.

[0075] Step 3: Strain screening based on ecological network standards.

[0076] The CarveMe tool was used to construct genome-scale metabolic models from the high-quality MAGs obtained in step 1 above. The PhyloMint method was used to quantify the pairwise metabolic complementarity indices, generating an adjacency matrix. Interactions with metabolic complementarity indices less than 2.698 in the adjacency matrix were filtered using the Z-score outlier detection method, resulting in the final adjacency matrix used for constructing the metabolic complementarity network. The intra-module connectivity (Zi) and inter-module connectivity (Pi) of each node in the network were calculated. Core nodes with Zi ≥ 2.5 or / and Pi ≥ 0.62 were selected as key species in the activated sludge microbial metabolic complementarity network (see reference). Figure 2 ).

[0077] Step 4: Take the intersection of the various strains obtained from Steps 1 to 3 to obtain strains N, G, H and D.

[0078] like Figure 3 As shown, the degradation abilities of strains N, G, and H on SMX and the degradation ability of strain D on 1,2,4-phenylpyrogallol were verified. Figure 3 The bar chart represents the concentration of remaining sulfamethoxazole (initial concentration of 1 ug / L), and the scatter plot represents the concentration of 1,2,4-phenylpyrogallol produced by strains N, G, and H, and the concentration of remaining 1,2,4-phenylpyrogallol (initial concentration of 1 ug / L) corresponding to strain D.

[0079] Strains N, G, and H were inoculated into LB medium, with an initial SMX concentration of 1 μg / L for strains N, G, and H. After pure culture at room temperature with shaking for 72 h, the degradation rates of SMX by N, G, and H were 91.1%, 86.5%, and 88.2%, respectively, and the product 1,2,4-pyrogallol was detected. It should be noted that the LB medium described above is a commonly used technique in this field, and the formulation of the LB medium in the experimental examples of this application is not limited.

[0080] For strain D, strain D was inoculated into LB medium with an initial concentration of 1,2,4-phenylpyrogallol of 1 μg / L. After pure culture at room temperature with shaking for 72 h, the degradation rate of 1,2,4-phenylpyrogallol was 94.7%.

[0081] Understandably, the above steps did not detect β-ketoadipic acid because it is a natural intermediate metabolite, not a pollutant, and non-toxic. Furthermore, in major databases such as KEGG, MetaCyc, and BRENDA, all annotated degradation pathways of 1,2,4-phenylpyrogallol point to β-ketoadipic acid, and no other complete, functionally validated alternative pathways were found. Therefore, it can be assumed that the degradation product of 1,2,4-phenylpyrogallol is β-ketoadipic acid, and detection is unnecessary.

[0082] Therefore, it can be seen that strains N, G, and H all have the ability to degrade SMX into 1,2,4-pyrogallol, while strain D has the ability to completely degrade 1,2,4-pyrogallol.

[0083] The following describes the verification process of the synthetic microbial community's degradation effect on SMX in wastewater as described in Examples 1-7.

[0084] (1) Validation of SMX intake flux.

[0085] The microbial community metabolic model MICOM was used to predict the uptake flux of SMX by the seven synthetic bacterial groups corresponding to Examples 1 to 7.

[0086] Specifically, the seven synthetic bacterial groups mentioned above correspond to seven combinations: combination 1 is G+D, combination 2 is H+D, combination 3 is N+D, combination 4 is G+H+D, combination 5 is G+N+D, combination 6 is H+N+D, and combination 7 is G+H+N+D. For example... Figure 4 As shown, combinations 4 and 7 exhibited the highest flux and were selected as candidate combinations.

[0087] Among them, combination 1 is the synthetic microbial community of Example 1, combination 2 is the synthetic microbial community of Example 2, combination 3 is the synthetic microbial community of Example 3, combination 4 is the synthetic microbial community of Example 4, combination 5 is the synthetic microbial community of Example 5, combination 6 is the synthetic microbial community of Example 6, and combination 7 is the synthetic microbial community of Example 7.

[0088] (2) Laboratory pure culture verification Candidate combinations were inoculated into LB medium, supplemented with 1 μg / L SMX, and cultured with shaking at room temperature (25℃). The concentration change of SMX was monitored by high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS). The results showed that the SMX concentration decreased significantly, and after 72 hours, the removal rates of SMX by combinations 4 and 7 were both higher than 83% (reference). Figure 5 Based on the combined experimental data and the highest throughput, combination 7 was determined to be the optimal combination.

[0089] (3) Real wastewater environmental verification The optimal combination 7 is divided into 5×10 7 Cells / mL were inoculated into activated sludge collected from a municipal wastewater treatment plant. The influent contained 1 μg / L SMX. After 7 days of operation, the SMX removal rate of the sequencing batch reactor reached 65.5%. Figure 6 ).

[0090] In summary, the working principle of the above seven synthetic microbial communities provided in the embodiments of this application is based on a synergistic mechanism of functional division of labor, quorum sensing, and ecological colonization.

[0091] In terms of functional division of labor, the first degrading bacteria (N, G, H) preferentially contact SMX and convert it into intermediate metabolites through their respective initial degrading enzyme systems; the second degrading bacteria D specifically utilizes this intermediate for subsequent degradation, forming a "relay" metabolic chain, reducing the metabolic burden of individual strains.

[0092] Quorum sensing coordination: Individual strains in the synthetic microbial community sense community density in real time by secreting and sensing self-induced signal molecules. Furthermore, bacteria with quorum sensing capabilities can respond to changes in the external environment in real time, maintaining the overall stability of the microbial community.

[0093] Ecological colonization guarantee: Since all strains in the synthetic microbial community are core species in the activated sludge metabolic network (high Zi and / or Pi), their metabolic needs are highly compatible with the indigenous microorganisms in the wastewater treatment system, and they can coexist with the indigenous bacteria, thus maintaining the degradation function of SMX in the long term.

[0094] It is understood that the various embodiments in this specification are described in a progressive manner, and the same or similar parts between the various embodiments can be referred to each other. Each embodiment focuses on describing the differences from other embodiments.

[0095] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A synthetic microbial community, characterized in that, It includes a first degrading functional bacterium and a second degrading functional bacterium, wherein the first degrading functional bacterium includes Nocardiopsis coralliicola , Gordoniahydrophobica , Halioglobus pacificus At least one of the bacteria, wherein the second degrading bacteria includes Dokdonella fugitiva .

2. The synthetic microbial community as described in claim 1, characterized in that, The ratio of viable first-degrading functional bacteria to second-degrading functional bacteria in the synthetic microbial community is 0.2~5:

1.

3. A synthetic microbial community as described in claim 2, characterized in that, The first degrading functional bacteria include Nocardiopsis coralliicola , Gordoniahydrophobica , Halioglobus pacificus When two types of bacteria are present, the live bacteria ratio of the two types of bacteria is 0.01 to 100:

1.

4. A synthetic microbial community as described in claim 2, characterized in that, The first degrading functional bacteria include Nocardiopsis coralliicola , Gordoniahydrophobica as well as Halioglobus pacificus At that time, the Nocardiopsis coralliicola , Gordoniahydrophobica as well as Halioglobus pacificus The live bacteria ratio is 0.01~100:0.01~100:

1.

5. The application of the synthetic microbial community as described in any one of claims 1 to 4 in the degradation of sulfamethoxazole in wastewater.

Images