Bioactive hybrid micro-robot for removing antibiotic pollution of water body and preparation method and application of bioactive hybrid micro-robot

Abstract

The invention discloses a bioactive hybrid micro-robot for removing antibiotic pollution of a water body as well as a preparation method and application of the bioactive hybrid micro-robot, and belongs to the technical field of water pollution treatment. Chlamydomonas reinhardtii is coupled with DBCO-PEG4-NHS, then copper-free click chemistry (SPAAC) and p-azidobenzoic acid (AzBA) react for further assembly modification, functionalized algae are further assembled into the micro-robot, and the micro-robot achieves rapid adsorption of SMX by means of the stable self-driving characteristic of the algae; in the whole reaction process, through the synergistic effect of hydrogen bonds and pi bonds between p-azidobenzoic acid and SMX, dynamic removal of SMX in the environmental water body is finally achieved. According to the invention, the biological hybrid micro-robot (AzBA-algae robot) with functional activity is developed, efficient removal of SMX is realized, and as a technical method meeting ecological requirements, the technology opens up a new path for removal of SMX in an environmental water sample, and also provides an innovative thought for dynamic removal research of other types of environmental pollutants.

Description

TECHNICAL FIELD

[0001] The application relates to a bioactive hybrid microrobot for removing antibiotic pollution in a water body as well as a preparation method and application thereof, and belongs to the technical field of treatment of antibiotic pollution in water. BACKGROUND

[0002] In the past decade, antibiotics have attracted much attention as a new type of pollutant. The reason is that antibiotics are continuously released and widely exist in aquatic environments, and organisms contacting such compounds may pose potential risks to aquatic ecosystems and human health. Sulfonamide antibiotics (including sulfamethoxazole, SMX) are widely used in human medical treatment and livestock breeding fields and have been popularized due to their broad-spectrum antibacterial activity, economy and easy synthesis. However, their large-scale use has also caused serious environmental problems: due to the incomplete metabolism of organisms to such compounds, most sulfonamide antibiotics (such as SMX) are discharged into aquatic systems in the form of original or metabolite, leading to continuous pollution of water bodies. The accumulation of SMX in the environment can cause multiple risks to aquatic ecosystems and human health, including the induction of antibiotic resistance and potential carcinogenicity - the World Health Organization has made relevant risk classification.

[0003] Removing SMX from the water environment has become an urgent environmental problem. Traditional treatment technologies (such as activated sludge method) have limited efficiency in removing SMX, mainly due to the poor biodegradability of SMX and the inhibitory effect on microbial activity. The activated carbon physical adsorption method has great potential for SMX removal, but this method has defects such as high operating cost and the need for complex equipment support. Chemical oxidation technology can effectively degrade SMX, but it may produce toxic by-products or require a large amount of energy input. These inherent limitations highlight the urgency of developing innovative, efficient and environmentally friendly SMX removal technologies.

[0004] With the development of environmental remediation technology, micro robots have the ability to move autonomously, can increase the collision contact frequency with target pollutants, achieve autonomous mixing of local water bodies, and shorten the pollution removal period, becoming a new direction for dynamic removal of water environmental pollutants. Previous studies have shown that such active micro robots have shown advantages in the removal of various pollutants (such as heavy metals, viruses, etc.). However, the current micro robot system for water environment remediation still has many technical defects, which limits its large-scale application. Specifically, the effective working time is short, and the remediation function cannot be maintained for a long time; some rely on toxic driving fuel, which is easy to cause secondary pollution and violates biological safety requirements; complex external driving equipment (such as magnetic field, light control device, etc.) is needed, which is difficult to operate and has poor adaptability; the applicable water environment range is narrow, and it can only work in single-purity water (such as ultrapure water), and it is difficult to adapt to actual complex water environments (such as high-turbidity river water, impurity-containing aquaculture wastewater). SUMMARY

[0005] The application aims to provide a bioactive hybrid microrobot for removing antibiotic pollution in water bodies, and provide a simple, compatible and biologically safe technical solution for dynamic removal of sulfamethoxazole in water environment. The microrobot can also efficiently purify various common pollutants in water bodies due to its excellent autonomous movement ability and flexible surface functional modification characteristics, thereby providing a dynamic and efficient solution path for water environment management.

[0006] To achieve the above technical purpose, the application adopts the following technical solution: A bioactive hybrid microrobot for removing antibiotic pollution in water bodies, comprising Chlamydomonas reinhardtii and p-azidobenzoic acid modified on the surface of the algae by DBCO-PEG4-NHS reagent coupling.

[0007] Preferably, the antibiotic is sulfamethoxazole.

[0008] The application further provides a preparation method of the bioactive hybrid microrobot for removing antibiotic pollution in water bodies, comprising the following steps: (1) Culturing Chlamydomonas reinhardtii: Chlamydomonas reinhardtii is cultured in TAP medium at room temperature for 24-36 h, the algal cells are collected by centrifugal separation, and the Chlamydomonas reinhardtii is obtained by centrifugal washing with ultrapure water; (2) The Chlamydomonas reinhardtii cells obtained in step (1) are resuspended with ultrapure water and mixed with DBCO-PEG4-NHS solution and cultured at a specific temperature for 1-2 h, the algal cells are collected by centrifugal separation and washed with ultrapure water to obtain DBCO-modified algal cells; (3) The obtained DBCO-modified algal cells are resuspended with ultrapure water and mixed with p-azidobenzoic acid solution and incubated at room temperature for 1-1.5 h.

[0009] Preferably, the volume ratio of the Chlamydomonas reinhardtii cell suspension to the DBCO-PEG4-NHS solution in step (2) is 1:1; the concentration of the DBCO-PEG4-NHS solution is 20 μM, and the cell concentration of the Chlamydomonas reinhardtii cell suspension is 1.0×10 7 / mL.

[0010] Preferably, the culture temperature is 23-25℃.

[0011] Preferably, the volume ratio of the DBCO-modified algal cell suspension to the p-azidobenzoic acid solution in step 3 is 1:1; the concentration of the p-azidobenzoic acid solution is 20 μM; and the cell concentration of the DBCO-modified algal cell suspension is 1.0×10 7 / mL.

[0012] Preferably, the antibiotic is sulfamethoxazole.

[0013] The application further provides application of the above-mentioned bioactive hybrid microrobot for removing antibiotic pollution in a water body, which is used for removing sulfamethoxazole in the water body. The microrobot maintains a movement speed of about 11.3 μm / s in eight kinds of aqueous media (comparable to unmodified algae), and can still maintain stable activity for 24 hours in a solution containing 0.5 mg / L SMX, and exhibits excellent environmental adaptability. In actual tests, the SMX removal rate of the microrobot in ultrapure water within 2 hours reaches 83.5% (only 14.7% for unmodified algae), and the removal rate in six actual water samples reaches 84.1% to 87.7%. As an economical, efficient and environmentally friendly biological hybrid system, the technology not only solves the SMX repair problem, but also provides a scalable solution for dynamic removal of various pollutants in the water body.

[0014] Compared with the prior art, the application has the following beneficial effects: 1) The bioactive hybrid microrobot of the application accelerates purification through dynamic movement to achieve rapid adsorption equilibrium, realizes efficient removal of SMX, and shortens the purification period.

[0015] 2) The core material of the application is safe, non-toxic, non-toxic and non-residual, has excellent biocompatibility, and has no risk of any secondary pollution.

[0016] 3) The application has strong environmental adaptability, is widely applicable, can be compatible with various water quality environments, and is resistant to SMX pollution environment; and the preparation process is simple, batch production is possible, the detection and maintenance cost is low, and industrial application is easy. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 The figure is a mechanism diagram of the bioactive hybrid microrobot for removing antibiotic pollution in a water body of the application; Figure 2 The figure is a laser confocal fluorescence characterization of the bioactive hybrid microrobot for removing antibiotic pollution in a water body of the application and an infrared spectrum of related substances, wherein A is a bright field image (BF), a fluorescence image and a merged image of the functionalized algal microrobot: the Cy5 channel shows the autofluorescence of natural algal chlorophyll, and the FITC channel shows the fluorescence signal of FAM; B is an infrared characterization spectrum of DBCO-PEG4-NHS, p-azidobenzoic acid and the click reaction product of the two; Figure 3 The figure is functionalization modification of algae and its influence on movement behavior; wherein A is a comparison of movement speeds of unmodified algae, DBCO modified algae and AzBA-algal robots; B, C and D are movement trajectory diagrams (scale: 5 μm) of the three kinds of algae respectively; E is a longitudinal space scanning image of the movement trajectory of Chlamydomonas reinhardtii in the functionalization modification process under different Z-stack distances; Figure 4Figure 1. The motion performance of AzBA-algobot in different water environments; Wherein A-F are the representative 2-second optical motion trajectories of 0-hour functionalized algomicrobots in different aqueous media: A drinking water, B lake water, C river water, D effluent from sewage treatment plant, E aquaculture water in Shu River, F water in poultry breeding area of lake region; G-L are the 2-second motion trajectories in the corresponding media for 24 hours (the order is the same as above); M is the motion characteristic analysis of AzBA-algobot in different water environments: 0.1x phosphate buffered saline (PBS), triacetate phosphate medium (TAP), drinking water, lake water, river water, effluent from sewage treatment plant, aquaculture water in Shu River, water in poultry breeding area of lake region (scale: 5 pm); Figure 5 Figure 2. The motion performance of AzBA-algobot for removing SMX. Wherein A is drinking water, B is lake water, C is river water, D is effluent from sewage treatment plant, E is aquaculture water in Shu River, F is water in poultry breeding area of lake region (scale: 5 pm); Figure 6 Figure 3. The removal effect of AzBA-algobot on sulfamethoxazole (SMX). Wherein A is the removal kinetics curve of AzBA-algobot and unmodified algae on SMX in ultrapure water; B is the time-dependent efficiency comparison of AzBA-algobot and unmodified algae for removing SMX (n=3 independent repeated experiments); Figure 7 Figure 4. The SMX removal effect of AzBA-algobot in actual water samples. Wherein A is the removal rate of SMX in different actual water samples; B-G is the concentration comparison before and after SMX removal in actual water samples (the order is the same as above) (n=3 independent repeated experiments). DETAILED DESCRIPTION

[0018] The technical solutions of the present application are further described below in conjunction with specific embodiments, but are not limited thereto. Some reagents used in the present application are sourced as follows: sulfamethoxazole (SMX) is purchased from Shanghai Mailin Biotechnology Co., Ltd.; p-azidobenzoic acid (AzBA) is purchased from Shanghai Mailin Biotechnology Co., Ltd.; 5-isomer arylamine azide is purchased from Shanghai Jinpan Biological Technology Co., Ltd.; 4% paraformaldehyde fixing solution and PBS buffer are both purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.; Chlamydomonas reinhardtii strain and TAP medium are provided by the National Aquatic Biological Germplasm Repository; ultrapure water (UP water) used in the experiment is prepared by the laboratory ultrapure water system. Water samples include: commercially available drinking pure water (Hangzhou Wahaha Group), effluent from sewage treatment plant in Luozhuang District, Linyi City, river water in Shu River, Linyi County, lake water in Hongshi Lake, Linyi County, aquaculture-affected water in Shu River, and water discharged from poultry breeding farm in Linyi County.

[0019] The water samples used in the experiment include tap water, lake water, drinking water, effluent from a sewage treatment plant, aquaculture water in a marsh, and water from a poultry and livestock breeding area in a lake region. All water samples were treated according to the following steps: First, sulfamethoxazole was added to each water sample to prepare a drug-containing water sample; 500 mL of the prepared water sample was filtered through a 0.22 μm filter membrane to remove suspended particulate matter and microbial contamination; the filtrate was stored at 4°C and analyzed within 24 hours to avoid degradation of sulfamethoxazole.

[0020] SMX was dissolved in deionized water to prepare a stock solution, and the desired concentration of working solution was obtained by gradient dilution of the stock solution. In the experiment, 500 μL of SMX working solution with a concentration of 0.5 mg / L was accurately transferred into the reaction system containing functionalized algal micro-robots, and incubated at 23°C after thorough mixing. After incubation, the sample was centrifuged at low speed for 3 minutes, and the supernatant was filtered through a 0.22 μm nylon filter membrane into a sample bottle. The concentration of SMX in the filtrate was determined by high performance liquid chromatography (HPLC).

[0021] The concentration of antibiotics was determined using a Waters HPLC system equipped with an automatic sampler and a UV detector, with a detection wavelength of 254 nm. A Lotus C18 chromatographic column (5.0 μm, Chromai) was used to ensure high separation performance. The mobile phase consisted of two parts: mobile phase A was pure acetonitrile (100%), and mobile phase B was a mixture of ultrapure water and acetic acid (99.9:0.1 by volume). The elution process used isocratic elution mode, with a volume ratio of mobile phase A to B set at 50:50, and a flow rate controlled at 1 mL·min -1 The sample analysis was performed with an injection volume of 20 μL to ensure the accuracy and reproducibility of the test results.

[0022] Example 1 Preparation of biologically active hybrid micro-robots for removing antibiotic pollution in water bodies In this study, copper-free click chemistry (SPAAC) strategy was used to realize the surface functionalization of algae. The specific steps are as follows: (1) Cultivation of Chlamydomonas reinhardtii: Chlamydomonas reinhardtii was cultured in TAP medium at room temperature for 24-36 h, and the cells were collected by centrifugation at 3000 rpm for 3 minutes and washed with ultrapure water for 3 times. The purpose was to completely remove the residual culture medium components and avoid interference of impurities in the subsequent DBCO modification reaction; (2) The Chlamydomonas reinhardtii cells obtained in step (1) were resuspended with DBCO-PEG4-NHS solution and incubated at 23-25°C for 1-2 h. Then the algal cells were separated by centrifugation for 3 min and washed with ultrapure water to obtain DBCO-modified algal cells; (3) The DBCO-modified algal cells obtained are resuspended in ultrapure water and mixed with the p-azidobenzoic acid solution at room temperature for 1-1.5 h to obtain the product.

[0023] The volume ratio of the Chlamydomonas reinhardtii cell suspension to the DBCO-PEG4-NHS solution in step (2) is 1:1; the concentration of the DBCO-PEG4-NHS solution is 20 μM, and the cell concentration of the Chlamydomonas reinhardtii cell suspension is 1.0×10 7 individuals / mL.

[0024] The volume ratio of the DBCO-modified algal cell suspension to the p-azidobenzoic acid solution in step 3 is 1:1; the concentration of the p-azidobenzoic acid solution is 20 μM; and the cell concentration of the DBCO-modified algal cell suspension is 1.0×10 7 individuals / mL.

[0025] In the above preparation method, first, the extracellular polymers of the algal cells are chemically modified by using the DBCO-PEG4-NHS reagent. The N-hydroxysuccinimide (NHS) ester group in the DBCO-PEG4-NHS molecule undergoes nucleophilic substitution reaction with the amino or hydroxyl groups on the surface of the algal cells, and the dibenzocyclooctyne (DBCO) group is covalently coupled to the surface of the algal extracellular polymers. As a high-efficiency bioorthogonal reaction module, the DBCO group can form a stable triazole bond with a molecule containing an azido group through a cycloaddition reaction without a copper catalyst.

[0026] The molecular design of DBCO-PEG4-NHS has unique advantages: the DBCO part as a cyclooctyne derivative provides a high-efficiency click reaction site, and the PEG4 spacer significantly improves the water solubility and biocompatibility of the reagent, allowing it to react gently with biological components in a water environment while minimizing the impact on the activity of the algal cells.

[0027] In the subsequent modification step, the DBCO-modified algal cells are incubated with p-azidobenzoic acid at room temperature for 1 hour. Under this condition, the azido group in the p-azidobenzoic acid is rapidly coupled with the DBCO group on the surface of the algal cells through copper-free click chemistry, forming a stable covalent bond. This reaction has high selectivity and efficiency, which can ensure the site-specific fixation of p-azidobenzoic acid on the surface of the algal cells.

[0028] Example 2 Test and characterization of the bioactive hybrid microrobot prepared in the present application To verify the success of the bioactive hybrid microrobot prepared in the present application, this example carried out a fluorescence co-localization experiment, and the results are as follows: Figure 2As shown in Figure A, the morphology and distribution characteristics of the algae were clearly presented in bright-field (BF) optical microscopy images by utilizing the autofluorescence properties of Chlamydomonas reinhardtii chloroplasts and combining them with spatial localization analysis of the azide-labeled FAM fluorescent probe. The autofluorescence of chloroplasts detected in the Cy5 channel highly overlapped with the bright-field image, confirming the endogenous origin of the fluorescence signal. In the FITC channel, FAM fluorescence was uniformly distributed on the algal surface and spatially overlapped with Cy5 chloroplast fluorescence. The merged images directly demonstrate that the azide-modified FAM fluorescent probe was successfully anchored to the microalgal surface through click chemistry, providing intuitive evidence for the effectiveness of functionalization modification.

[0029] Infrared spectroscopy further confirmed the success of the functionalization modification, specifically manifested in the shift of characteristic peaks ( Figure 2 (B) In the Fourier transform infrared (FT-IR) spectrum after the click reaction, the terminal azido group shows a characteristic absorption peak at 2100 cm⁻¹; when DBCO participates in the reaction and successfully binds to p-azidobenzoic acid, a reflection absorption Fourier transform infrared (RA-FTIR) spectrum appears at 1530 cm⁻¹. -1 1440 cm -1 and 1047 cm -1 Three absorption peaks correspond to the vibrational signals of the 1,2,3-triazole ring. Since the formation of this ring depends on the click reaction between DBCO and the azide group, this indirectly proves that DBCO participated in the reaction and that the click reaction occurred successfully.

[0030] Example 3 Motion performance of the bioactive hybrid microrobot prepared in this invention in ultrapure water medium AzBA-algal robots were prepared by incubating DBCO-modified Chlamydomonas reinhardtii with p-azidobenzoic acid (AzBA) for 1 hour.

[0031] The movement trajectory of the AzBA-algal microrobot in ultrapure water was recorded using an inverted fluorescence microscope (NIKON TI-E). The stability of its movement speed was monitored at 0 h and 24 h to ensure its autonomous movement capability in complex aquatic environments. Results are shown below. Figure 3 .from Figure 3 Data analysis showed that the movement speeds of the microrobots in DBCO-modified and p-azidobenzoic acid-modified algae were 11.7 μm / s and 11.3 μm / s, respectively, which were comparable to the movement speed of unmodified *Chlamydomonas reinhardtii* (11.75 μm / s), indicating that the functionalization process had minimal impact on the algal movement ability. Figure 3 (A) Figure 3The study primarily showcased the activity status of unmodified *Chlamydomonas reinhardtii*, DBCO-modified algae, and p-azidobenzoic acid-modified algal microrobots after 24 hours of incubation in ultrapure water. Their activity characteristics were assessed by recording the movement trajectories of the three groups of algae. The results showed that the activity status of DBCO-modified algae and functionalized algal microrobots was not significantly different from that of unmodified *Chlamydomonas reinhardtii*, further confirming that the effect of functionalization modification on algal activity is negligible.

[0032] In addition, Z-stack tomography was performed on functionalized Chlamydomonas reinhardtii using laser confocal scanning microscopy to construct its distribution modification trajectory. Figure 3 (E), the results further confirmed that the functionalization process had a weak effect on algal survival.

[0033] Example 4 The bioactive hybrid microrobot prepared in this invention exhibits improved mobility in different aqueous media. The AzBA algal robot was placed in dishes along with different aqueous media, and its movement trajectory in the different aqueous media was recorded using an inverted fluorescence microscope (NIKONTI-E). Figure 4 As shown, the AzBA-algae robot prepared in this invention exhibited continuous motion capability in all tested aqueous media. Figure 4 The microrobot's stable motion performance was demonstrated by its representative 2-second motion trajectories in drinking water, lake water, river water, sewage treatment plant effluent, Shuhe River aquaculture water, and lake area livestock farming water. This continuous motion capability was verified through 0 hours ( Figure 4 (A, B, C, D, E, F) and 24 hours ( Figure 4 The motion trajectories of G, H, I, J, K, and L in all the aforementioned media were further verified. Notably, the AzBA-algal robot prepared in ultrapure water maintained a self-driving speed of 11.5-12.5 μm / s in eight different aquatic media, including 0.1×PBS, TAP culture medium, drinking water, lake water, river water, wastewater treatment plant effluent, Shuhe River aquaculture water, and water from livestock and poultry farming areas in the lake region. Figure 4 (M).

[0034] Example 5 The activity of AzBA-algal robots obtained in this invention in sulfamethoxazole solution In this embodiment, the bioactivity and motion stability of the AzBA-algal robot in a sulfamethoxazole (SMX) contaminated environment were systematically evaluated. Figure 5Representative 2-second motion trajectories of 20 AzBA-algal robots after 24 hours of exposure in various aquatic environments were presented: drinking water (A), lake water (B), river water (C), wastewater treatment plant effluent (D), aquaculture water from the Shuhe River (E), and water from a livestock and poultry farming area in a lake region (F). The results showed that the presence of 0.5 mg / L SMX in all test media did not adversely affect the survival time and motility of the AzBA-algal robots. Crucially, the AzBA-algal robots maintained their full operational performance, exhibiting stable motility even after 24 hours, with no observed reduction in survival time. The trajectory stability of this bio-hybrid system remained consistently high, confirming its sustained motility capability in complex polluted environments.

[0035] Example 6 The removal efficiency of sulfamethoxazole by the AzBA-algal robot obtained in this invention The removal efficiency of AzBA-algal robot for sulfamethoxazole (SMX) was determined by systematic adsorption experiments. To quantify the effect of azidobenzoic acid modification on the SMX removal efficiency, unmodified algae (ordinary microalgae without functionalization) and AzBA-algal robot were added to ultrapure water with an SMX concentration of 0.5 mg / L, respectively. The results showed that AzBA-algal robot reached adsorption equilibrium within 2 hours, with an SMX removal rate of 83.5%. Figure 6 (Blue curve in A); Under the same experimental conditions, the SMX removal rate of unmodified algae was only 14.7% ( Figure 6 The red curve in section A confirms that the removal efficiency of functionalized microrobots (FMRs) is significantly higher than that of unmodified algae.

[0036] Example 7 Application of the AzBA-algae robot obtained in this invention in actual aquatic environments To verify the application effectiveness of the AzBA-algae robot in actual aquatic environments, multi-media adsorption performance tests were conducted. The AzBA-algae robot was incubated with various actual water samples (drinking water, lake water, river water, wastewater treatment plant effluent, Shuhe River aquaculture water, and water from a livestock and poultry farming area in the lake) containing 0.5 mg / L SMX at 23±2℃ for 2 hours. The SMX concentration in the supernatant was then analyzed by HPLC. The results showed that the SMX removal rate in all tested water bodies reached 84.1%–87.7%. Figure 7 (A) Figure 7 Further analysis by B confirmed that the residual SMX concentration in the treated samples was significantly lower than the initial concentration. These data collectively demonstrate that the AzBA-algal robot possesses stable and efficient SMX removal capabilities in both artificial systems and complex environmental matrices, confirming its good applicability in various aquatic systems.

[0037] Based on the above experimental results, this invention verifies the successful development of an active biohybrid AzBA-algal robot for the dynamic remediation of sulfamethoxazole (SMX). The *Chlamydomonas reinhardtii* was functionalized with azide benzoic acid (AzBA) using copper-free click chemistry (SPAAC) technology, enabling the robot to bind to SMX through hydrogen bonds and π-π interactions (verified by DFT calculations). This robot maintained a movement speed of approximately 11.3 μm / s in eight different aqueous media (comparable to unmodified algae), and maintained stable activity for 24 hours in solutions containing 0.5 mg / L SMX, demonstrating strong environmental adaptability. In ultrapure water, the robot achieved an SMX removal rate of 83.5% within 2 hours (compared to only 14.7% for unmodified algae); the removal rate ranged from 84.1% to 87.7% in six actual water samples. As an economical, efficient, and environmentally friendly system, this technology not only solves the SMX remediation problem but also provides an innovative strategy for the dynamic removal of various pollutants in water bodies that can be applied on a large scale.

[0038] It should be noted that the above embodiments are merely some preferred embodiments of the present invention, and not all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

Claims

1. A bioactive hybrid micro-robot for removing antibiotic pollution from water bodies, characterized by, It comprises Chlamydomonas reinhardtii and p-azidobenzoic acid modified on the surface of the algae by DBCO-PEG4-NHS reagent coupling.

2. The bioactive hybrid micro-robot for removing antibiotic contamination in water bodies as claimed in claim 1, wherein, The antibiotic is sulfamethoxazole.

3. A method for preparing the bioactive hybrid microrobot for removing antibiotic pollution in water bodies according to claim 1 or 2, characterized in that, The method comprises the following steps: (1) Culturing Chlamydomonas reinhardtii: Chlamydomonas reinhardtii is cultured in TAP medium at room temperature for 24-36 h, and then the algae cells are collected by centrifugal separation and washed with ultrapure water to obtain Chlamydomonas reinhardtii; (2) The Chlamydomonas reinhardtii cells obtained in step (1) are resuspended with ultrapure water and mixed with DBCO-PEG4-NHS solution and cultured at a specific temperature for 1-2 h, and then the algae cells are collected by centrifugal separation and washed with ultrapure water to obtain DBCO-modified algae cells; (3) The obtained DBCO-modified algae cells are resuspended with ultrapure water and mixed with p-azidobenzoic acid solution and incubated at room temperature for 1-1.5 h to obtain the product.

4. The method for preparing bioactive hybrid microrobots for removing antibiotic pollution from water bodies according to claim 3, characterized in that, The volume ratio of the Chlamydomonas reinhardtii cell resuspension solution to the DBCO-PEG4-NHS solution in the step (2) is 1:1; the concentration of the DBCO-PEG4-NHS solution is 20 μM, and the cell concentration of the Chlamydomonas reinhardtii cell resuspension solution is 1.0×10 7 8 / mL.

5. The method for preparing bioactive hybrid microrobots for removing antibiotic pollution from water bodies according to claim 3, characterized in that, The culture temperature is 23-25°C.

6. The method for preparing bioactive hybrid microrobots for removing antibiotic pollution from water bodies according to claim 3, characterized in that, The volume ratio of the DBCO-modified algal cell resuspension in step 3 to the solution of p-azidobenzoic acid is 1:1; the concentration of the solution of p-azidobenzoic acid is 20 μM; and the cell concentration of the DBCO-modified algal cell resuspension is 1.0 x 10 7 8 cells / mL.

7. The method for preparing bioactive hybrid microrobots for removing antibiotic pollution from water bodies according to claim 3, characterized in that, The antibiotic is sulfamethoxazole.

8. Use of the bioactive hybrid microrobot for removing antibiotic pollution from water bodies according to claim 1 or 2, characterized in that, The method is used for removing sulfamethoxazole in water. The method is used for removing sulfamethoxazole in water.

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