Bioactive hybrid microrobot for removing antibiotic pollution in water body and preparation method and application thereof

Abstract

This invention discloses a bioactive hybrid microrobot for removing antibiotic pollution from water bodies, its preparation method, and its application, belonging to the field of water pollution treatment technology. This invention couples *Chlamydomonas reinhardtii* with DBCO-PEG4-NHS, and then further assembles and modifies it through copper-free click chemistry (SPAAC) and a reaction with p-azidobenzoic acid (AzBA), further assembling the functionalized algae into microrobots. Relying on the stable self-driving characteristics of the algae, the microrobots achieve rapid adsorption of antibiotics (SMX). Throughout the reaction process, the synergistic effect of hydrogen bonds and π bonds between p-azidobenzoic acid and SMX ultimately achieves the dynamic removal of SMX from environmental water bodies. This invention develops a functionally active bioactive hybrid microrobot (AzBA-algal robot) that achieves highly efficient SMX removal. As an ecologically sound technical method, this technology opens up a new pathway for the removal of SMX from environmental water samples and provides innovative ideas for the dynamic removal research of other types of environmental pollutants.

Description

Technical Field

[0001] This invention relates to a bioactive hybrid microrobot for removing antibiotic pollution from water, its preparation method and application, belonging to the field of antibiotic pollution treatment technology in water. Background Technology

[0002] Over the past decade, antibiotics have emerged as a new type of pollutant, drawing significant attention. This is due to their continuous release and widespread presence in aquatic environments, posing potential risks to aquatic ecosystems and human health upon exposure to these compounds. Sulfonamide antibiotics (including sulfamethoxazole (SMX)) are widely used in human medicine and livestock farming, gaining popularity due to their broad-spectrum antibacterial activity, affordability, and ease of synthesis. However, their extensive use has also caused serious environmental problems: because organisms do not fully metabolize these compounds, most sulfonamide antibiotics (such as SMX) are released into aquatic systems either unchanged or as metabolites, leading to persistent water pollution. The accumulation of SMX in the environment poses multiple risks to aquatic ecosystems and human health, including inducing antibiotic resistance and potential carcinogenicity—a risk classification established by the World Health Organization.

[0003] Removing solid methane (SMX) from aquatic environments has become an urgent environmental problem. Traditional treatment technologies (such as activated sludge processes) have limited removal efficiency for SMX, mainly because SMX has poor biodegradability and inhibits microbial activity. Activated carbon physical adsorption shows great potential for SMX removal, but this method suffers from drawbacks such as high operating costs and the need for complex equipment. While chemical oxidation technologies can effectively degrade SMX, they may produce toxic byproducts or require significant energy input. These inherent limitations highlight the urgent need to develop innovative, efficient, and environmentally friendly SMX removal technologies.

[0004] With the development of environmental remediation technologies, microrobots, possessing autonomous mobility, can shorten pollutant removal cycles by increasing the frequency of collisions with target pollutants and achieving autonomous mixing in localized water bodies, thus becoming a new direction for the dynamic removal of pollutants from aquatic environments. Previous studies have confirmed that these active microrobots have demonstrated advantages in the removal of various pollutants (such as heavy metals and viruses). However, current microrobot systems used for aquatic environment remediation still have many technical shortcomings, limiting their large-scale application. These include: short effective working time, unable to maintain remediation functions for extended periods; partial reliance on toxic driving fuels, easily causing secondary pollution and violating biosafety requirements; the need for complex external driving equipment (such as magnetic fields and light control devices), resulting in high operational difficulty and poor adaptability; and a narrow range of applicable aquatic environments, only able to operate in water bodies of single purity (such as ultrapure water), making it difficult to adapt to actual complex aquatic environments (such as highly turbid river water and aquaculture wastewater containing impurities). Summary of the Invention

[0005] The purpose of this invention is to provide a bioactive hybrid microrobot for removing antibiotic pollution from water bodies, offering a simple, compatible, and biosafe technical solution for the dynamic removal of sulfamethoxazole from aquatic environments. These microrobots, with their excellent autonomous movement capabilities and flexible surface functional modification properties, can also efficiently purify various common pollutants in water bodies, thus providing a dynamic and efficient solution for water environment management.

[0006] To achieve the above-mentioned technical objectives, the present invention adopts the following technical solution:

[0007] A bioactive hybrid microrobot for removing antibiotic pollution from water bodies comprises Chlamydomonas reinhardtii and p-azidobenzoic acid coupled to the surface of the algae via DBCO-PEG4-NHS reagent.

[0008] Preferably, the antibiotic is sulfamethoxazole.

[0009] The present invention also provides a method for preparing the above-mentioned bioactive hybrid microrobot for removing antibiotic pollution from water, comprising the following steps:

[0010] (1) Cultivation of Chlamydomonas reinhardtii: Chlamydomonas reinhardtii was cultured in TAP medium at room temperature for 24-36 h, the algal cells were collected by centrifugation and washed with ultrapure water by centrifugation to obtain Chlamydomonas reinhardtii;

[0011] (2) The Chlamydomonas reinhardtii cells obtained in step (1) were resuspended in ultrapure water and then mixed with DBCO-PEG4-NHS solution and cultured at a specific temperature for 1-2 h. The algal cells were collected by centrifugation and washed with ultrapure water to obtain DBCO-modified algal cells.

[0012] (3) The obtained DBCO-modified algal cells were resuspended in ultrapure water and mixed with p-azidobenzoic acid solution and incubated at room temperature for 1-1.5 h to obtain the final product.

[0013] Preferably, in step (2), the volume ratio of the Chlamydomonas reinhardtii cell resuspension to the DBCO-PEG4-NHS solution is 1:1; the concentration of the DBCO-PEG4-NHS solution is 20 μM, and the cell concentration of the Chlamydomonas reinhardtii cell resuspension is 1.0 × 10⁻⁶. 7 per mL.

[0014] Preferably, the culture temperature is 23-25℃.

[0015] Preferably, in step 3, the volume ratio of the DBCO-modified algal cell resuspension to the p-azidobenzoic acid solution is 1:1; the concentration of the p-azidobenzoic acid solution is 20 μM; and the cell concentration of the DBCO-modified algal cell resuspension is 1.0 × 10⁻⁶ cells / mL. 7 per mL.

[0016] Preferably, the antibiotic is sulfamethoxazole.

[0017] This invention also provides an application of the aforementioned bioactive hybrid microrobot for removing antibiotic pollution from water, specifically for the removal of sulfamethoxazole from water. This microrobot maintains a movement speed of approximately 11.3 μm / s in eight different aqueous media (comparable to unmodified algae), and retains stable activity for 24 hours in a solution containing 0.5 mg / L SMX, demonstrating excellent environmental adaptability. In actual tests, the microrobot achieved an SMX removal rate of 83.5% in ultrapure water within 2 hours (compared to only 14.7% in unmodified algae), and a removal rate of 84.1%–87.7% in six different actual water samples. As an economical, efficient, and environmentally friendly bio-hybrid system, this technology not only solves the SMX remediation problem but also provides a scalable solution for the dynamic removal of various pollutants in water.

[0018] Compared with the prior art, the beneficial effects of the present invention are:

[0019] 1) The bioactive hybrid microrobot of the present invention achieves rapid adsorption equilibrium through dynamic motion to accelerate purification, thereby realizing efficient removal of SMX and shortening the purification cycle.

[0020] 2) The core materials of this invention are safe and non-toxic, with no toxic byproducts or residues, excellent biocompatibility, and no risk of secondary pollution.

[0021] 3) This invention has strong environmental adaptability, wide applicability, is compatible with various water quality environments, and is resistant to SMX pollution environments; moreover, the preparation process is simple, it can be mass-produced, the testing and maintenance costs are low, and it is easy to apply industrially. Attached Figure Description

[0022] Figure 1 This is a diagram illustrating the mechanism of action of the bioactive hybrid microrobot for removing antibiotic pollution from water bodies according to the present invention.

[0023] Figure 2 This image shows the laser confocal fluorescence characterization of the bioactive hybrid microrobot for removing antibiotic pollution from water bodies according to the present invention, and the infrared spectra of related substances. A represents the bright field image (BF), fluorescence image, and 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 represents the infrared characterization spectra of DBCO-PEG4-NHS, p-azidobenzoic acid, and their click reaction products.

[0024] Figure 3This study presents the functional modification of algae and its impact on their movement behavior. A shows a comparison of the movement speeds of unmodified algae, DBCO-modified algae, and AzBA-algal robot. B, C, and D are the movement trajectory diagrams of the three algae (scale bar: 5 μm). E is a longitudinal spatial scan image of the movement trajectory of Chlamydomonas reinhardtii during the functional modification process at different Z-stack distances.

[0025] Figure 4 This invention presents the motion performance of the AzBA-algae robot in different aquatic environments. AF represents the representative 2-second optical motion trajectory of the functionalized algae microrobot in different aquatic media at 0 hours: A - drinking water, B - lake water, C - river water, D - wastewater treatment plant effluent, E - Shuhe River aquaculture water, F - lake area livestock and poultry farming water; GL represents the 2-second motion trajectory in the corresponding media at 24 hours (in the same order); M represents the motion characteristic analysis of the AzBA-algae robot in different aquatic environments: 0.1× phosphate-buffered saline (PBS), triacetic acid phosphate medium (TAP), drinking water, lake water, river water, wastewater treatment plant effluent, Shuhe River aquaculture water, lake area livestock and poultry farming water (scale bar: 5 μm).

[0026] Figure 5 The representative 2-second motion trajectory of the AzBA-algae robot after removing SMX is shown, where A is drinking water, B is lake water, C is river water, D is sewage treatment plant effluent, E is Shuhe River aquaculture water, and F is lake area livestock and poultry farming water (scale bar: 5 μm).

[0027] Figure 6 The removal efficiency of AzBA-algal robot for sulfamethoxazole (SMX) is shown in Figure A, where A represents the removal kinetic curves of SMX by AzBA-algal robot and unmodified algae in ultrapure water; and B represents the time-dependent efficiency comparison of SMX removal by AzBA-algal robot and unmodified algae (n=3 independent replicate experiments).

[0028] Figure 7 The SMX removal effect of AzBA-algae robot in actual water samples is shown in Figure A, where A represents the SMX removal rate in different actual water samples; and B and G represent the concentration comparison of SMX before and after removal in actual water samples (in the same order as above) (n=3 independent replicate experiments). Detailed Implementation

[0029] The technical solution of the present invention will be further described below with reference to specific embodiments, but is not limited thereto. Some of the reagents used in this invention were sourced from the following sources: sulfamethoxazole (SMX) was purchased from Shanghai Mailin Biochemical Technology Co., Ltd.; p-azidobenzoic acid (AzBA) was purchased from Shanghai Maclean Biochemical Technology Co., Ltd.; 5-isomer aromatic amine azide was purchased from Shanghai Jinpan Biotechnology Co., Ltd.; 4% paraformaldehyde fixative and PBS buffer were both purchased from Sinopharm Chemical Reagent Co., Ltd.; Chlamydomonas reinhardtii strain and TAP medium were provided by the National Aquatic Organism Germplasm Resource Bank; the ultrapure water (UP water) used in the experiment was prepared by the laboratory ultrapure water system. Water samples included: commercially available drinking water (Hangzhou Wahaha Group), effluent from the Luozhuang District Sewage Treatment Plant in Linyi City, Shuhe River water in Linshu County, Hongshi Lake water in Linshu County, water affected by aquaculture in the Shuhe River fish farm, and effluent from livestock farms in Linshu County.

[0030] The water samples used in the experiment included tap water, lake water, drinking water, effluent from a sewage treatment plant, aquaculture water from the Shuhe River, and water from livestock and poultry farming areas in the lake area. All water samples were processed 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℃ and analyzed within 24 hours to avoid degradation of sulfamethoxazole.

[0031] SMX was dissolved in deionized water to prepare a stock solution, which was then serially diluted to obtain the working solution of the desired concentration. In the experiment, 500 μL of a 0.5 mg / L SMX working solution was precisely transferred into the reaction system containing the functionalized algal microrobots, thoroughly mixed, and incubated at 23 °C. After incubation, the sample was centrifuged at low speed for 3 minutes, and the supernatant was filtered through a 0.22 μm nylon membrane into a sample vial. The concentration of SMX in the filtrate was determined by high-performance liquid chromatography (HPLC).

[0032] Antibiotic concentrations were determined using a Waters HPLC system equipped with an autosampler and a UV detector, with the detection wavelength set at 254 nm. Chromatographic separation was performed using a Lotus C18 column (5.0 μm, Chromai) to ensure high separation efficiency. 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 (volume ratio 99.9:0.1). Elution was performed in isocratic elution mode, with the volume ratio of mobile phase A to B set at 50:50, and the flow rate controlled at 1 mL / min. -1 The method runs for 10 minutes. During sample analysis, the injection volume is precisely controlled at 20 μL to ensure the accuracy and repeatability of the results.

[0033] Example 1

[0034] Preparation of bioactive hybrid microrobots for removing antibiotic pollution from water bodies

[0035] This study employed a copper-free click chemistry (SPAAC) strategy to achieve surface functionalization modification of algae. The specific steps were as follows:

[0036] (1) Cultivation of Chlamydomonas reinhardtii: Chlamydomonas reinhardtii was cultured in TAP medium at room temperature for 24-36 h. Cells were collected by centrifugation at 3000 rpm for 3 minutes and washed 3 times with ultrapure water. The purpose was to thoroughly remove residual culture medium components and avoid impurities from interfering with the subsequent DBCO modification reaction.

[0037] (2) The Chlamydomonas reinhardtii cells obtained in step (1) were resuspended in ultrapure water and mixed with DBCO-PEG4-NHS solution and cultured at 23-25℃ for 1-2 h. After that, the algal cells were separated and collected by centrifugation for 3 min and washed with ultrapure water to obtain DBCO-modified algal cells.

[0038] (3) The obtained DBCO-modified algal cells were resuspended in ultrapure water and mixed with p-azidobenzoic acid solution and incubated at room temperature for 1-1.5 h to obtain the final product.

[0039] In step (2), the volume ratio of *Chlamydomonas reinhardtii* cell resuspension to DBCO-PEG4-NHS solution is 1:1; the concentration of DBCO-PEG4-NHS solution is 20 μM, and the cell concentration of *Chlamydomonas reinhardtii* cell resuspension is 1.0 × 10⁻⁶. 7 per mL.

[0040] In step 3, the volume ratio of the DBCO-modified algal cell resuspension to the p-azidobenzoic acid solution is 1:1; the concentration of the p-azidobenzoic acid solution is 20 μM; and the cell concentration of the DBCO-modified algal cell resuspension is 1.0 × 10⁻⁶ cells / mL. 7 per mL.

[0041] In the above preparation method, the extracellular polymeric material of algal cells is first chemically modified using the DBCO-PEG4-NHS reagent. The N-hydroxysuccinimide (NHS) ester group in the DBCO-PEG4-NHS molecule undergoes a nucleophilic substitution reaction with the amino or hydroxyl groups on the surface of the algal cells, covalently coupling the dibenzocyclooctylene (DBCO) group to the surface of the algal extracellular polymeric material. As a highly efficient bioorthogonal reaction module, the DBCO group can form stable triazole bonds with molecules containing azide groups through cycloaddition reactions under copper-free conditions.

[0042] The molecular design of DBCO-PEG4-NHS has unique advantages: the DBCO part, as a cyclooctyne derivative, provides a highly efficient click reaction site, while the PEG4 spacer arm significantly improves the water solubility and biocompatibility of the reagent, enabling it to react mildly with biological components in an aquatic environment, while minimizing the impact on algal cell activity.

[0043] In the subsequent modification step, DBCO-modified algal cells were incubated with p-azidobenzoic acid at room temperature for 1 hour. Under these conditions, the azido groups in p-azidobenzoic acid rapidly coupled with the DBCO groups on the algal cell surface via copper-free click chemistry, forming stable covalent bonds. This reaction exhibits high selectivity and efficiency, ensuring the site-specific fixation of p-azidobenzoic acid on the algal cell surface.

[0044] Example 2

[0045] Test and characterization of the bioactive hybrid microrobots prepared in this invention

[0046] To verify the successful fabrication of the bioactive hybrid microrobot prepared in this invention, a fluorescence co-localization experiment was conducted in this embodiment. The results are as follows: Figure 2 As 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.

[0047] 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.

[0048] Example 3

[0049] Motion performance of the bioactive hybrid microrobot prepared in this invention in ultrapure water medium

[0050] AzBA-algal robots were prepared by incubating DBCO-modified Chlamydomonas reinhardtii with p-azidobenzoic acid (AzBA) for 1 hour.

[0051] 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 3 The 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.

[0052] 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.

[0053] Example 4

[0054] The bioactive hybrid microrobot prepared in this invention exhibits improved mobility in different aqueous media.

[0055] 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).

[0056] Example 5

[0057] The activity of AzBA-algal robots obtained in this invention in sulfamethoxazole solution

[0058] In this embodiment, the bioactivity and motion stability of the AzBA-algal robot in a sulfamethoxazole (SMX) contaminated environment were systematically evaluated. Figure 5 Representative 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.

[0059] Example 6

[0060] The removal efficiency of sulfamethoxazole by the AzBA-algal robot obtained in this invention

[0061] 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.

[0062] Example 7

[0063] Application of the AzBA-algae robot obtained in this invention in actual aquatic environments

[0064] 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 area) containing 0.5 mg / L SMX at 23±2℃ for 2 hours. Subsequently, the SMX concentration in the supernatant was 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.

[0065] 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.

[0066] 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 biologically active hybrid micro-robot for removing antibiotic pollution from a water body, characterized in that, It comprises Chlamydomonas reinhardtii and p-azidobenzoic acid modified on the surface of the algae by DBCO-PEG4-NHS reagent coupling; the antibiotic is sulfamethoxazole; The preparation method of the bioactive hybrid microrobot for removing antibiotic pollution in a water body comprises the following steps: (1) Culturing Chlamydomonas reinhardtii: Chlamydomonas reinhardtii is cultured in a TAP medium at room temperature for 24-36 h, the algal cells are collected by centrifugal separation, and the Chlamydomonas reinhardtii cells are obtained by centrifugal washing with ultrapure water; (2) The Chlamydomonas reinhardtii cells obtained in step (1) are resuspended with ultrapure water, mixed with a DBCO-PEG4-NHS solution, and cultured at 23-25 DEG C for 1-2 h, the algal cells are collected by centrifugal separation, and the DBCO-modified algal cells are obtained by centrifugal washing with ultrapure water; (3) The DBCO-modified algal cells are resuspended with ultrapure water, mixed with a p-azidobenzoic acid solution, and incubated at room temperature for 1-1.5 h, and the bioactive hybrid microrobot for removing antibiotic pollution in a water body is obtained.

2. The bioactive hybrid micro-robot for removing antibiotic contamination in water bodies according to claim 1, wherein, The volume ratio of the Chlamydomonas reinhardtii cell suspension obtained after resuspending the Chlamydomonas reinhardtii cells in ultrapure water in step (2) to the DBCO-PEG4-NHS solution 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 8 / mL.

3. The bioactive hybrid microrobot for removing antibiotic contamination in a water body according to claim 1, characterized in that, The volume ratio of the DBCO-modified algal cell suspension obtained after the DBCO-modified algal cells in step (3) were resuspended with ultrapure water to the p-azidobenzoic acid solution 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 x 10 7 9 / mL.

4. Use of the bioactive hybrid microrobot for removing antibiotic pollution from water bodies as claimed in claim 1, wherein, The bioactive hybrid microrobot is used for removing sulfamethoxazole in a water body.

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