Conjugated microporous polymer-Chlorella complex, its preparation method and its application in treating organic pollutants in wastewater.

Abstract

This invention relates to the field of wastewater treatment, and particularly to a conjugated microporous polymer-Chlorella complex, its preparation method, and its application in treating organic pollutants in wastewater. The preparation method includes: cultivating Chlorella in a culture medium to obtain a Chlorella culture solution; centrifuging the Chlorella culture solution to collect Chlorella cells; adding a sulfone-functionalized conjugated microporous polymer to the culture medium and sonicating it to obtain a conjugated microporous polymer suspension; steps b) and c) are not sequentially restricted; mixing the Chlorella cells with the conjugated microporous polymer suspension to obtain a composite solution; and incubating the composite solution to obtain the conjugated microporous polymer-Chlorella complex. The conjugated microporous polymer-Chlorella complex prepared by this method has a significant effect on removing organic pollutants from water bodies, providing a new solution for water pollution control.

Description

Technical Field

[0001] This invention relates to the field of wastewater treatment, and more particularly to conjugated microporous polymer-Chlorella complex, its preparation method, and its application in treating organic pollutants in wastewater. Background Technology

[0002] With the acceleration of industrialization and the development of agricultural activities, organic pollutants in water bodies, including aniline, pesticides, and drug residues, pose a serious threat to the ecological environment and human health. These organic pollutants are chemically stable and difficult to biodegrade, making traditional physical sedimentation, chemical oxidation, and biological treatment methods often ineffective and potentially leading to secondary pollution. Among biological treatment methods, microalgae are considered a promising biological purifier due to their efficient photosynthesis and strong pollutant absorption capacity. The oxygen produced by microalgae through photosynthesis can promote the aerobic degradation of organic pollutants, and the enzyme system within microalgal cells can also directly participate in the metabolic process of pollutants. However, microalgae face many limitations in practical applications. For example, the efficiency of photosynthesis is affected by light intensity and spectral quality, limiting the degradation rate and efficiency of organic pollutants. Furthermore, the activity and stability of microalgae are difficult to guarantee under complex aquatic environmental conditions. To improve the pollutant degradation capacity of microalgae, researchers have attempted to modify them using physical, chemical, or bioengineering techniques. For example, genetic engineering can be used to increase the expression levels of specific enzymes within microalgal cells, or physical methods can be used to enhance the adsorption capacity of microalgae for pollutants. Although these methods have improved the purification efficiency of microalgae to some extent, they still face problems such as high cost, complex operation, and difficulty in large-scale application. Summary of the Invention

[0003] To address or partially address the problems existing in related technologies, this invention provides a conjugated microporous polymer-Chlorella complex, its preparation method, and its application in treating organic pollutants in wastewater.

[0004] This invention provides a method for preparing a conjugated microporous polymer-Chlorella complex, comprising:

[0005] Step a) Chlorella is cultured in a culture medium to obtain Chlorella culture solution;

[0006] Step b) Centrifuge the Chlorella culture medium to collect the Chlorella cells;

[0007] Step c) The sulfone-functionalized conjugated microporous polymer is added to the culture medium and subjected to ultrasonic treatment to obtain a conjugated microporous polymer suspension; there is no order restriction between steps b) and c).

[0008] Step d) The Chlorella cells are mixed with a conjugated microporous polymer suspension to obtain a composite solution;

[0009] Step e) Incubate the composite liquid to obtain a conjugated microporous polymer-Chlorella complex.

[0010] Furthermore, the sulfone-functionalized modified conjugated microporous polymer is prepared according to the following method:

[0011] Under nitrogen protection and high-temperature calcination, using Pd2(dba)3 and tris(o-methoxyphenyl)phosphine as catalysts and ligands, monomers TBrPy and DTDO undergo cross-coupling reactions in the presence of the catalyst. Simultaneously, the disulfide bonds in DTDO are oxidized to sulfone groups, ultimately forming a conjugated microporous polymer modified with sulfone functional groups.

[0012] Furthermore, the Chlorella is Chlorella proteoglycans.

[0013] Furthermore, the concentration of the sulfone-functionalized conjugated microporous polymer in the composite solution is 0.1-0.5 g / L; the absorbance OD of the composite solution at 680 nm in the UV-Vis spectrum is... 680 It is 0.4-0.8.

[0014] Further, the culture medium is sterile BG-11 medium; the sterile BG-11 medium comprises: 1.5 g / L sodium nitrate, 0.075 g / L magnesium sulfate, 0.006 g / L citric acid, 0.04 g / L potassium dihydrogen phosphate, 0.006 g / L sodium iron citrate, 0.02 g / L sodium carbonate, 0.036 g / L calcium chloride, 0.001 g / L ethylenediaminetetraacetic acid, and 1 ml / L of a trace mineral mixture; the trace mineral mixture comprises: 0.049 g / L cobalt nitrate, 0.079 g / L copper sulfate, 0.222 g / L zinc sulfate, 0.079 g / L sodium molybdate, 1.81 g / L manganese chloride, and 2.86 g / L boric acid.

[0015] Furthermore, step a) is specifically as follows:

[0016] Chlorella was cultured in the sterile BG-11 medium using a cold white fluorescent light source with an intensity of 4000 Lux and a photoperiod of 14 hours followed by 10 hours of darkness. The temperatures during the light and dark phases were maintained at 28°C and 24°C, respectively. Cell density was determined by measuring the absorbance (OD680) at 680 nm using UV-Vis spectroscopy. When the OD680 reached 0.4-0.8, the cells were removed and used to obtain the Chlorella culture medium.

[0017] Furthermore, in step b), the centrifugation speed is 5000-8000 rpm and the time is 5-10 min.

[0018] Furthermore, in step e), the incubation temperature is 28-30℃, the incubation time is 60-90 min, and the stirring speed is 110-180 rpm.

[0019] The present invention also provides a conjugated microporous polymer-Chlorella complex, which is prepared according to any one of the methods described above.

[0020] The present invention also provides an application of the above-mentioned conjugated microporous polymer-Chlorella complex in the treatment of organic pollutants in wastewater.

[0021] The conjugated microporous polymer-Chlorella complex and its preparation method provided by this invention can have the following beneficial effects:

[0022] 1. The preparation method involves uniformly mixing Chlorella with a sulfone-functionalized conjugated microporous polymer under physical stirring conditions. The electrostatic and orbital interactions between Chlorella and the conjugated microporous polymer drive their composite formation, thus obtaining a Chlorella-conjugated microporous polymer composite.

[0023] 2. The prepared composite, under light stimulation, can enhance the photosynthetic systems PS1 and PS2 of Chlorella and energy metabolism processes such as oxidative phosphorylation through photoelectrons generated by the conjugated microporous polymer, thereby increasing the production of ATP, NADPH, and NADH by Chlorella. This composite, through the synergistic effect of photosynthesis and biological metabolism, reduces energy consumption and operating costs during the treatment process.

[0024] 3. Compared with Chlorella alone, the prepared composite improves the degradation efficiency of Chlorella for organic pollutants, achieving rapid removal of pollutants and exhibiting superior removal effect on aniline pollutants. It shows promise for the degradation of recalcitrant organic pollutants. The introduction of sulfone functional groups enhances the biocompatibility and environmental adaptability of the composite, broadening its application scope.

[0025] 4. Experimental results show that the sulfone-functionalized conjugated microporous polymer-Chlorella complex has a significant effect on removing organic pollutants from water bodies, providing a new solution for water pollution control.

[0026] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and are not intended to limit the invention. Attached Figure Description

[0027] The above and other objects, features and advantages of the present invention will become more apparent from the more detailed description of exemplary embodiments of the invention in conjunction with the accompanying drawings, wherein the same reference numerals generally represent the same parts.

[0028] Figure 1 These are SEM images of the sulfone-functionalized conjugated microporous polymer and Chlorella vulgaris in Example 1 of this invention, as well as SEM images and TEM slices of the conjugated microporous polymer-Chlorella vulgaris complex.

[0029] Figure 2 This is a laser confocal image of the conjugated microporous polymer-Chlorella complex in Example 1 of the invention;

[0030] Figure 3 This is a graph showing the NADH, NADPH, and ATP contents of Chlorella and the conjugated microporous polymer-Chlorella complex in Example 1 of the invention.

[0031] Figure 4 These are graphs showing the effect of different test substances in removing aniline contaminants in Example 1 of the present invention;

[0032] Figure 5 This is a kinetic curve of the removal of aniline contaminants by different test substances in Example 1 of the present invention. Detailed Implementation

[0033] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0034] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The singular forms “a,” “the,” and “the” used in this invention and the appended claims are also intended to include the plural forms unless the context clearly indicates otherwise. It should also be understood that the term “and / or” as used herein refers to and includes any or all possible combinations of one or more of the associated listed items.

[0035] It should be understood that although the terms "first," "second," "third," etc., may be used in this invention to describe various information, this information should not be limited to these terms. These terms are only used to distinguish information of the same type from one another. For example, without departing from the scope of this invention, first information may also be referred to as second information, and similarly, second information may also be referred to as first information. Thus, features defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, "a plurality of" means two or more, unless otherwise explicitly specified.

[0036] To address the aforementioned issues, the inventors of this application considered introducing nanomaterials into microalgae-based pollutant degradation technology. Conjugated microporous polymers (CMPs) are photosensitive organic semiconductor polymers that exhibit great potential in pollutant adsorption and catalytic degradation due to their unique light absorption properties, high specific surface area, and designable pore structure. Therefore, the inventors considered combining CMPs with microalgae to improve the pollutant degradation capacity of microalgae. However, unlike the matrix of general composite materials, microalgae are biological organisms. Therefore, the combination of CMPs, as non-biological materials, with microalgae faces problems such as poor biocompatibility and loose contact.

[0037] Furthermore, the inventors of this application considered functional group modification of the conjugated microporous polymer to improve its binding performance with microalgae. The inventors discovered that conjugated microporous polymers modified with sulfone functional groups can enhance hydrophilicity and charge transfer efficiency with the Chlorella cell membrane, promoting the adsorption and internalization of organic pollutants by microalgae. In addition, by regulating the charge distribution inside and outside the microalgal cells through the electronic properties of the sulfone functional groups, the electron transport chain is optimized, improving the photosynthetic efficiency and energy metabolism of microalgae. Therefore, this application ultimately combines the sulfone-functionalized conjugated microporous polymer with Chlorella. The resulting composite not only improves the photosynthetic efficiency of microalgae but also enhances their metabolic activity and pollutant degradation capacity by promoting electron transfer, achieving highly efficient removal of organic pollutants from water.

[0038] This invention provides a method for preparing a conjugated microporous polymer-Chlorella complex, comprising:

[0039] Step a) Chlorella is cultured in a culture medium to obtain Chlorella culture solution;

[0040] Step b) Centrifuge the Chlorella culture medium to collect the Chlorella cells;

[0041] Step c) The sulfone-functionalized conjugated microporous polymer is added to the culture medium and subjected to ultrasonic treatment to obtain a conjugated microporous polymer suspension; there is no order restriction between steps b) and c).

[0042] Step d) The Chlorella cells are mixed with a conjugated microporous polymer suspension to obtain a composite solution;

[0043] Step e) Incubate the composite liquid to obtain a conjugated microporous polymer-Chlorella complex.

[0044] In the above preparation method, step a) is the process of cultivating and proliferating Chlorella to obtain a Chlorella culture medium with a certain cell density. The cultivated Chlorella provides raw materials for subsequent steps. As a preferred embodiment, the Chlorella is preferably Chlorella pyrenoidosa. Chlorella pyrenoidosa Specifically, it can be the Chlorella proteoglycans FACHB-9 from the Institute of Hydrobiology, Chinese Academy of Sciences.

[0045] In this step, the culture medium used for cultivating Chlorella is preferably sterile BG-11 medium. Specifically, the sterile BG-11 medium includes: 1.5 g / L sodium nitrate, 0.075 g / L magnesium sulfate, 0.006 g / L citric acid, 0.04 g / L potassium dihydrogen phosphate, 0.006 g / L sodium iron citrate, 0.02 g / L sodium carbonate, 0.036 g / L calcium chloride, 0.001 g / L ethylenediaminetetraacetic acid, and 1 ml / L of a trace mineral mixture. The trace mineral mixture includes: 0.049 g / L cobalt nitrate, 0.079 g / L copper sulfate, 0.222 g / L zinc sulfate, 0.079 g / L sodium molybdate, 1.81 g / L manganese chloride, and 2.86 g / L boric acid.

[0046] Furthermore, this step is preferably implemented as follows:

[0047] Chlorella was cultured in the sterile BG-11 medium using a cold white fluorescent light source with an intensity of 4000 Lux, and a photoperiod of 14 hours followed by 10 hours of darkness. The temperatures for the light and dark phases were maintained at 28°C and 24°C, respectively. Cell density was determined by measuring the absorbance (OD680) at 680 nm using UV-Vis spectroscopy. Cells were removed from the medium when the OD680 reached 0.4–0.8, yielding the Chlorella culture medium. More preferably, the Chlorella culture medium was removed from the medium when the OD680 reached 0.4–0.8.

[0048] Steps b) and c) above are steps for obtaining Chlorella cells and sulfone-functionalized conjugated microporous polymer suspensions, respectively. There is no restriction on the order of these two steps, and they can be performed simultaneously. Step b) will be described in detail first, followed by a detailed description of step c).

[0049] Step b) above involves separating Chlorella cells by centrifugation to remove impurities such as the culture medium, thus obtaining pure Chlorella cells. This ensures the effective binding of Chlorella cells to CMPs in subsequent steps. In this step, the preferred centrifugation speed is 5000-8000 rpm, and the preferred centrifugation time is 5-10 min. Most preferably, the centrifugation speed is 5000 rpm, and the time is 5 min. Pure Chlorella cells are obtained through step b).

[0050] Step c) above involves uniformly dispersing the sulfone-functionalized conjugated microporous polymer in the culture medium using ultrasound to provide uniform and suitable conditions for subsequent composite formation. In this step, the preferred ultrasound frequency is 50 W, and the preferred time is 10 min. The CMPs used in this step are sulfone-functionalized CMPs, whose sulfone functional groups enhance their hydrophilicity and charge transfer efficiency, making them easier to bind to the cell membrane of *Chlorella vulgaris*. Furthermore, the electronic properties of the sulfone functional groups can regulate the charge distribution inside and outside the microalgal cells, optimize the electron transport chain, and improve photosynthetic efficiency and energy metabolism. The sulfone-functionalized conjugated microporous polymer is preferably prepared according to the following method:

[0051] Under nitrogen protection and high-temperature calcination, using Pd2(dba)3 and tris(o-methoxyphenyl)phosphine as catalysts and ligands, monomers TBrPy and DTDO undergo cross-coupling reactions in the presence of the catalyst. Simultaneously, the disulfide bonds in DTDO are oxidized to sulfone groups, ultimately forming a conjugated microporous polymer modified with sulfone functional groups.

[0052] In this preparation method, TBrPy (bromobiphenylpyridine) and DTDO (di(4-methylthiophenyl)disulfide) are used as reactants. The disulfide bonds in DTDO can be converted to sulfone functional groups during the reaction. The preferred molar ratio of TBrPy to DTDO is 1:1. Tris(dibenzylacetone)dipalladium Pd2(dba)3 is used as a catalyst, and tris(o-methoxyphenyl)phosphine is used as a ligand; they jointly promote the cross-coupling reaction between TBrPy and DTDO. The preferred molar ratio of tris(o-methoxyphenyl)phosphine to Pd2(dba)3 is 2:1. The preferred molar ratio of TBrPy, DTDO, tris(o-methoxyphenyl)phosphine, and Pd2(dba)3 is 100:100:6:3.

[0053] The cross-coupling reaction is carried out under nitrogen protection and high-temperature calcination, preferably at 100°C for 72 hours. Anhydrous 1,2-dimethylbenzene can be added as a solvent during the reaction, with a TBrPy to anhydrous 1,2-dimethylbenzene ratio of 1 mol:1 L. To ensure anhydrous and oxygen-free conditions and thus the catalytic effect of the metal catalyst, repeated vacuuming and nitrogen purging, as well as freeze-vacuum-thaw cycles, can be used. Anhydrous K₂CO₃ and metavalerate are also preferably added during the reaction to promote its progress. The preferred molar ratio of metavalerate to anhydrous K₂CO₃ to TBrPy is 3:15:5.

[0054] Step d) above is used to mix Chlorella cells with a conjugated microporous polymer suspension to form a composite solution. This step initially achieves the binding of Chlorella cells and the conjugated microporous polymer. As a preferred embodiment, the concentration of the sulfone-functionalized conjugated microporous polymer in the composite solution is 0.1-0.5 g / L, and the absorbance OD of the composite solution at 680 nm in the UV-Vis spectrum is [not specified]. 680 The concentration is 0.4-0.8. More preferably, the concentration of the sulfone-functionalized conjugated microporous polymer modified in the composite solution is 0.1 g / L, and the absorbance OD of the composite solution at 680 nm in the UV-Vis spectrum is [missing value]. 680 It is 0.4.

[0055] Step e) above is the incubation step of the composite solution, which promotes the interaction between the conjugated microporous polymer and Chlorella cells, forming a stable composite. The preferred incubation conditions are as follows: incubation temperature of 28-30℃, time of 60-90 min, and stirring speed of 110-180 rpm. More preferably, the incubation temperature is 30℃, the time is 60 min, and the stirring speed is 180 rpm. After incubation, the conjugated microporous polymer-Chlorella composite is obtained.

[0056] Another embodiment of the present invention provides a conjugated microporous polymer-Chlorella complex, which is prepared according to the method described in the above embodiments. The conjugated microporous polymer-Chlorella complex comprises Chlorella cells and a conjugated microporous polymer modified with a sulfone functional group bound to the exterior of the Chlorella cell wall.

[0057] As can be seen from the above, the conjugated microporous polymer-Chlorella complex and its preparation method provided in the embodiments of the present invention have the following advantages:

[0058] 1. The preparation method involves uniformly mixing Chlorella with a sulfone-functionalized conjugated microporous polymer under physical stirring conditions. The electrostatic and orbital interactions between Chlorella and the conjugated microporous polymer drive their composite formation, thus obtaining a Chlorella-conjugated microporous polymer composite.

[0059] 2. The prepared composite, under light stimulation, can enhance the photosynthetic systems PS1 and PS2 of Chlorella and energy metabolism processes such as oxidative phosphorylation through photoelectrons generated by the conjugated microporous polymer, thereby increasing the production of ATP, NADPH, and NADH by Chlorella. This composite, through the synergistic effect of photosynthesis and biological metabolism, reduces energy consumption and operating costs during the treatment process.

[0060] 3. Compared with Chlorella alone, the prepared composite improves the degradation efficiency of Chlorella for organic pollutants, achieving rapid removal of pollutants and exhibiting superior removal effect on aniline pollutants. It shows promise for the degradation of recalcitrant organic pollutants. The introduction of sulfone functional groups enhances the biocompatibility and environmental adaptability of the composite, broadening its application scope.

[0061] 4. Experimental results show that the sulfone-functionalized conjugated microporous polymer-Chlorella complex has a significant effect on removing organic pollutants from water bodies, providing a new solution for water pollution control.

[0062] Another embodiment of the present invention provides an application of the above-mentioned conjugated microporous polymer-Chlorella complex in the treatment of organic pollutants in wastewater. This conjugated microporous polymer-Chlorella complex combines the photosynthetic and metabolic pathways of microalgae to develop a novel biodegradation technology that converts organic pollutants into harmless inorganic substances or microbial biomass, thereby realizing the resource utilization of pollutants.

[0063] The technical solution of the present invention will be further described below with reference to specific embodiments:

[0064] Example 1

[0065] 1. Chlorella culture:

[0066] Chlorella strains Chlorella pyrenoidosa (FACHB-9) was purchased from the Institute of Hydrobiology, Chinese Academy of Sciences, and maintained in BG-11 medium. The medium consisted of: 1.5 g / L sodium nitrate, 0.075 g / L magnesium sulfate, 0.006 g / L citric acid, 0.04 g / L potassium dihydrogen phosphate, 0.006 g / L sodium iron citrate, 0.02 g / L sodium carbonate, 0.036 g / L calcium chloride, 0.001 g / L ethylenediaminetetraacetic acid, and 1 ml / L of a trace mineral mixture. The trace mineral mixture contained: 0.049 g / L cobalt nitrate, 0.079 g / L copper sulfate, 0.222 g / L zinc sulfate, 0.079 g / L sodium molybdate, 1.81 g / L manganese chloride, and 2.86 g / L boric acid.

[0067] Chlorella was cultured in 250 mL of sterile BG-11 medium. A cold white fluorescent light source with an intensity of 4000 Lux was used, with a 14-hour light cycle followed by a 10-hour dark cycle. The temperatures for the light and dark phases were maintained at 28°C and 24°C, respectively. All culture flasks were manually stirred three times daily and randomly placed to ensure single-cell culture. The absorbance (OD) at 680 nm was measured using UV-Vis spectroscopy. 680 ) to determine cell density. When OD 680 Once the concentration reaches 0.4, remove the culture medium and set it aside for later use to obtain Chlorella culture medium.

[0068] 2. Preparation of conjugated microporous polymers

[0069] First, 129.4 mg of TBrPy (0.5 mmol), 114.2 mg of DTDO (0.5 mmol), 30.6 mg of metavalerate (0.3 mmol), 207.3 mg of anhydrous K₂CO₃ (1.5 mmol), 10.6 mg of tris(o-methoxyphenyl)phosphine (0.03 mmol), and 13.7 mg of Pd₂(dba)₃ (0.015 mmol) were added sequentially to the Schlenk flask. The Schlenk flask was then repeatedly evacuated to remove air, and N₂ was introduced as a protective gas. Next, 5 mL of anhydrous 1,2-dimethylbenzene was slowly added to the solid mixture in the Schlenk flask using a syringe. To completely remove oxygen from the flask, the mixture was subjected to a freeze-evacuation-thawing cycle multiple times to ensure complete removal of oxygen. The solid mixture was then calcined at 100 °C for 72 h under a nitrogen atmosphere, with continuous stirring during the calcination process. After calcination, the solid mixture is allowed to cool naturally to room temperature. The precipitate is collected by centrifugation and finally dried to obtain a red powder, which is the sulfone-functionalized modified conjugated microporous polymer CMPs.

[0070] 3. Preparation of Chlorella-Conjugated Microporous Polymer Composite

[0071] Collect Chlorella cells from the Chlorella culture medium in step 1 by centrifugation at 5000 rpm for 5 minutes.

[0072] The sulfone-functionalized conjugated microporous polymer synthesized in step 2 was added to BG-11 medium with an equal volume of Chlorella culture medium and subjected to sonication at a frequency of 50 W for 10 min to obtain a 0.1 g / L conjugated microporous polymer suspension.

[0073] Collected Chlorella cells were simply mixed with a homogeneous suspension of conjugated microporous polymers at a concentration of 0.1 g / L to prepare a composite solution of Chlorella and CMPs. The concentration of the conjugated microporous polymers in the composite solution was 0.1 g / L, and the absorbance OD was [not specified].680 It is 0.4.

[0074] Subsequently, the composite solution was incubated at 30°C and 180 rpm for 60 min to promote the formation of the CMPs-Chlorella composite system. After incubation, the conjugated microporous polymer-Chlorella composite was obtained.

[0075] Figure 1 Image a shows a SEM image of the conjugated microporous polymer modified with the sulfone functional group described above; image b shows a SEM image of the collected Chlorella; image c shows a SEM image of the conjugated microporous polymer-Chlorella complex described above; and image d shows a TEM slice of the conjugated microporous polymer-Chlorella complex.

[0076] Scanning electron microscopy (SEM) analysis showed that the surface of Chlorella was smooth. Figure 1 (b) contrasts with the rough texture of the surface of the conjugated microporous polymer-Chlorella complex, which incorporates conjugated microporous polymers. Figure 1 c). Transmission electron microscopy (TEM) characterization clearly revealed that in the composite system, the aggregated conjugated microporous polymers were in close contact with Chlorella (c). Figure 1 d).

[0077] Figure 2 A laser confocal micrograph of the conjugated microporous polymer-Chlorella complex is shown. Under confocal laser scanning microscopy, the conjugated microporous polymer appears as a green pseudocolor, while individual Chlorella cells are characterized by a red pseudocolor. Figure 2 As expected, co-incubation of Chlorella with the conjugated microporous polymer resulted in well-positioned green pseudocolor on the Chlorella surface, coexisting with the inherent red pseudocolor, indicating that the conjugated microporous polymer has a strong affinity for the Chlorella surface. These observations suggest that the conjugated microporous polymer is located in the extracellular space.

[0078] [Detection of ATP, NADPH, and NADH]

[0079] The ATP, NADPH, and NADH levels in the Chlorella culture medium cultured alone and the conjugated microporous polymer-Chlorella complex in Example 1 were detected using the following methods:

[0080] Chlorella vulgaris and the complex formed by Chlorella vulgaris and conjugated microporous polymers (CMPs) were lysed by repeated freeze-thaw cycles. The mixture was then centrifuged at 5000 rpm for 5 minutes to obtain the supernatant. The total protein content was determined using the Coomassie Brilliant Blue assay (provided by Nanjing Jiancheng Biotechnology Institute). The contents of ATP, NADH, and NADPH were determined using a coenzyme I (NADH) assay kit and a coenzyme II (NADP / NADPH) assay kit provided by Nanjing Jiancheng Biotechnology Institute, and an enhanced ATP assay kit provided by Bio-Rad Laboratories.

[0081] The NADH, NADPH, and ATP contents of Chlorella and the conjugated microporous polymer-Chlorella complex are shown in the figure below. Figure 3 As shown, by Figure 3 The results showed that treatment with the conjugated microporous polymer significantly increased the contents of NADH and NADPH, reaching 0.022 μM and 0.058 μM, respectively, which were approximately 2.2 times and 1.3 times higher than those in Chlorella alone. Meanwhile, under light conditions, the ATP content in the conjugated microporous polymer-Chlorella mixture reached 0.61 μM, approximately 2.9 times that in Chlorella alone (0.21 μM).

[0082] [Pollutant Degradation Experiment]

[0083] A certain amount of aniline was introduced into different analytes and irradiated with a 300W xenon lamp for 4 hours. At specific time intervals, 1.5 mL of the reaction solution was taken out, filtered through a microfiltration membrane, and analyzed. The concentration of aniline was measured at 247 nm using a high-performance liquid chromatography (HPLC) system (Waters e2695, USA) equipped with a C18 column. A mixture of methanol and water was used as the mobile phase at a volume ratio of 60:40.

[0084] The substances to be tested are: the Chlorella culture medium obtained by culturing Chlorella alone in step 1 of Example 1, the conjugated microporous polymer suspension and the conjugated microporous polymer-Chlorella complex obtained in step 3.

[0085] The effect of different analytes on the removal of aniline contaminants and the corresponding kinetic curves are shown in the figure below. Figure 4 and Figure 5 As shown. By Figure 4 and Figure 5It was found that *Chlorella vulgaris* alone removed approximately 15% of aniline after 4 hours of light exposure, while the conjugated microporous polymer removed approximately 40% of aniline through photocatalysis. The conjugated microporous polymer-*Chlorella vulgaris* complex significantly outperformed *Chlorella vulgaris* alone, achieving a near 100% aniline removal rate, with a kinetic rate eight times that of *Chlorella vulgaris* alone. Under dark conditions, the aniline removal rate of the conjugated microporous polymer-*Chlorella vulgaris* complex was 20%, almost identical to the removal efficiency of *Chlorella vulgaris* alone, further confirming the importance of the photoelectrons of the conjugated microporous polymer in enhancing the metabolic activity of *Chlorella vulgaris*.

[0086] The various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or improvement of the technology in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing a conjugated microporous polymer-Chlorella complex, characterized in that, include: Step a) Chlorella is cultured in a culture medium to obtain Chlorella culture solution; Step b) Centrifuge the Chlorella culture medium to collect the Chlorella cells; Step c) The sulfone-functionalized modified conjugated microporous polymer is added to the culture medium and subjected to ultrasonic treatment to obtain a conjugated microporous polymer suspension; the sulfone-functionalized modified conjugated microporous polymer is prepared according to the following method: Under nitrogen protection and high-temperature calcination, using Pd2(dba)3 and tris(o-methoxyphenyl)phosphine as catalysts and ligands, monomers TBrPy and DTDO undergo cross-coupling reaction under the action of the catalyst. At the same time, the disulfide bond in DTDO is oxidized to sulfone group, and finally a conjugated microporous polymer modified with sulfone functional group is formed. There is no restriction on the order of steps b) and c). Step d) The Chlorella cells are mixed with a conjugated microporous polymer suspension to obtain a composite solution; Step e) Incubate the composite liquid to obtain a conjugated microporous polymer-Chlorella complex.

2. The preparation method according to claim 1, characterized in that, The Chlorella mentioned is Chlorella proteoglycans.

3. The preparation method according to claim 1, characterized in that, The concentration of the sulfone-functionalized conjugated microporous polymer in the composite solution is 0.1-0.5 g / L; the absorbance OD of the composite solution at 680 nm in the UV-Vis spectrum is... 680 It is 0.4-0.

8.

4. The preparation method according to claim 1, characterized in that, The culture medium is sterile BG-11 medium; the sterile BG-11 medium comprises: 1.5 g / L sodium nitrate, 0.075 g / L magnesium sulfate, 0.006 g / L citric acid, 0.04 g / L potassium dihydrogen phosphate, 0.006 g / L sodium iron citrate, 0.02 g / L sodium carbonate, 0.036 g / L calcium chloride, 0.001 g / L ethylenediaminetetraacetic acid, and 1 ml / L of a trace mineral mixture; the trace mineral mixture comprises: 0.049 g / L cobalt nitrate, 0.079 g / L copper sulfate, 0.222 g / L zinc sulfate, 0.079 g / L sodium molybdate, 1.81 g / L manganese chloride, and 2.86 g / L boric acid.

5. The preparation method according to claim 4, characterized in that, Step a) is as follows: Chlorella was cultured in sterile BG-11 medium using a cold white fluorescent light source at an intensity of 4000 Lux, with a 14-hour light cycle followed by a 10-hour dark cycle. The temperatures for the light and dark phases were maintained at 28°C and 24°C, respectively. The absorbance OD at 680 nm was measured using UV-Vis spectroscopy. 680 To determine the cell density; when the OD680 reaches 0.4-0.8, remove the cells for later use to obtain the Chlorella culture medium.

6. The preparation method according to claim 1, characterized in that, In step b), the centrifugation speed is 5000-8000 rpm and the time is 5-10 min.

7. The preparation method according to claim 1, characterized in that, In step e), the incubation temperature is 28-30℃, the incubation time is 60-90 min, and the stirring speed is 110-180 rpm.

8. A conjugated microporous polymer-Chlorella complex, characterized in that, It is prepared according to the method described in any one of claims 1 to 7.

9. The application of the conjugated microporous polymer-Chlorella complex according to claim 8 in the treatment of organic pollutants in wastewater.

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