Functional living body material prepared by adopting photosynthetic microcarrier and preparation method of functional living body material

Abstract

The invention provides a functional living body material prepared by adopting a photosynthetic microcarrier and a preparation method of the functional living body material. The preparation method comprises the following steps: respectively preparing first microbial microspheres encapsulated with microalgae and second microbial microspheres encapsulated with filamentous fungi by adopting a droplet microfluidic mode; after the first microbial microspheres and the second microbial microspheres are mixed to form a composite microcarrier, a functional living body material with a specific shape is constructed in a 3D printing mode, and the functional living body material is placed under a culture environment condition to be cultured. The microalgae as a photosynthetic active core can continuously perform photosynthesis under a suitable environmental condition, and the filamentous fungi enhance the stability of the microalgae through a supporting effect, so that the structural strength and long-term operability of the composite material are improved. Through the combination of the photosynthetic microcarriers, the functionality and operability of the material can be effectively improved.

Description

Technical Field

[0001] This application belongs to the field of biomaterial preparation and functionalized materials technology, specifically relating to a functional living material prepared using photosynthetic microcarriers and its preparation method. Background Technology

[0002] Functional living materials are produced through additive manufacturing, using cells as a substrate and biopolymers as carriers. Their functions originate from the physiological activities of cells and their synergistic effects with the biopolymers. These materials possess multiple functions, including self-repair, responsiveness, degradability, biosynthesis, pollution degradation, and carbon sequestration. They can also provide dynamic regulation through intelligent feedback and environmental adaptability, and are widely used in environmental remediation, medicine, energy production, and other fields.

[0003] However, functional living materials face challenges in their development, including poor stability, limited functionality, and insufficient adaptability. These problems mainly stem from their dependence on the physiological activities of organisms and external environmental conditions, resulting in a lack of sufficient stability and multifunctionality. Changes in the external environment, the limitations of the organism's life cycle, single-function design, and the lack of adaptive mechanisms restrict the widespread application of traditional living materials. Therefore, improving the stability, adaptability, and multifunctionality of living materials has become a major challenge for the industry's development. Summary of the Invention

[0004] To address the aforementioned issues, this application provides a functional living material prepared using photosynthetic microcarriers and its preparation method. By constructing photosynthetic microcarriers based on the symbiosis of filamentous fungal microspheres and microalgal microspheres, a functional living material with multiple functions, high stability, and excellent adaptability is designed and prepared.

[0005] To achieve the above objectives, one aspect of this application provides a method for preparing functional living materials using photosynthetic microcarriers, the method comprising the following steps: A first microbial microsphere and a second microbial microsphere were prepared using a droplet microfluidic method, wherein the first microbial microsphere encapsulated microalgae and the second microbial microsphere encapsulated filamentous fungi. The first microbial microspheres and the second microbial microspheres are mixed to form a composite microcarrier; The composite microcarrier is constructed into a functional living material with a specific shape by 3D printing and then placed in a culture environment for cultivation, so that the microalgae and the filamentous fungi form a symbiotic system.

[0006] In the functional living material prepared using photosynthetic microcarriers according to this application, microalgae serve as the photosynthetic active core, capable of continuously performing photosynthesis under suitable environmental conditions; filamentous fungi enhance the stability of the microalgae through a supporting role, improving the structural strength and long-term operability of the composite material. This combination of photosynthetic microcarriers effectively improves the functionality and operability of the material. This functional living material exhibits excellent biocompatibility and long-term stability, making it suitable for fields such as ecological restoration, resource recycling, and intelligent manufacturing, and possessing significant application value. This embodiment, by optimizing the preparation and application process of photosynthetic microcarriers, enhances the performance and practicality of the composite living material, demonstrating broad industrial application prospects.

[0007] Optionally, in conjunction with any of the above aspects, in another implementation of this aspect, the first microbial microspheres and the second microbial microspheres further include biopolymer materials, said biopolymer materials including one or more of thiolated hyaluronic acid (SH-HA), hyperbranched polyethylene glycol diacrylate (HB-PEGDA), thiolated sodium alginate (SH-SA), double-bonded gelatin (Gel-MA), and double-bonded F127 (F127-DA).

[0008] Alternatively, in conjunction with any of the above aspects, in another implementation of this aspect, the microalgae include one or more of Spirulina, Chlorella, cyanobacteria, genetically modified Spirulina, genetically modified Chlorella, and genetically modified cyanobacteria, which are capable of photosynthesis in a suitable culture environment.

[0009] Alternatively, in conjunction with any of the above aspects, in another implementation of this aspect, the filamentous fungi include Aspergillus niger, Polyporus genus, Sclerotium sclerotiorum, genetically engineered Aspergillus niger, or genetically engineered Polyporus genus.

[0010] Optionally, in conjunction with any of the above aspects, in another implementation of this aspect, the droplet microfluidic method includes: uniformly dispersing the microalgae or the filamentous fungi in the biopolymer material as a dispersed phase, dispensing the dispersed phase into microdroplets using continuous phase shearing, and then cross-linking and solidifying to form the first microbial microsphere and the second microbial microsphere.

[0011] Optionally, in conjunction with any of the above aspects, in another implementation of this aspect, the crosslinking and curing process includes any one of Michael addition reaction, photocrosslinking, esterification reaction, and ionic crosslinking, wherein the diameters of the first microbial microspheres and the second microbial microspheres are 100-300 μm, respectively.

[0012] Alternatively, in combination with any of the above aspects, in another implementation of this aspect, the first microbial microspheres and the second microbial microspheres are mixed at a volume ratio of 4:1 to form the composite microcarrier.

[0013] Optionally, in conjunction with any of the above aspects, in another implementation of this aspect, the culture environment conditions are a culture medium prepared by mixing BG11 and PDA in a 2:1 ratio, a temperature of 28-30°C, a light intensity of 5000-10000 Lux, a light-dark ratio of 12:12 hours, and a humidity of 50-60% RH.

[0014] Optionally, in conjunction with any of the above aspects, in another implementation of this aspect, the 3D printing method involves filling the composite microcarrier into a printing syringe and printing a scaffold model with a specific shape using a 3D printing device; wherein the 3D printing device includes any one of an extrusion printing device, an inkjet printing device, or a photopolymerization printing device. These additive manufacturing methods and technologies can precisely construct the microstructure and macroscopic morphology of functional living materials according to different needs.

[0015] Photosynthetic microcarriers are viscoelastic materials composed of hydrogel microspheres. When the volume fraction of microcarriers in a syringe exceeds 58%, a physical jamming effect occurs between the microspheres, causing the material to be in a solid state under low shear force. However, when the external force exceeds a certain level, the particles will slide and exhibit flow characteristics.

[0016] Another aspect of this application provides a functional living material prepared using photosynthetic microcarriers. The functional living material is prepared according to the method described in any of the above aspects. The microalgae in the functional living material produce oxygen and organic matter through photosynthesis, and the filamentous fungi secrete hyphae using carbon sources in the external environment and extend and grow along a preset printing path.

[0017] Furthermore, the functional living materials, through the precise combination of photosynthetic microcarriers, can significantly enhance the functionality, stability, and operability of the materials. This method enables the materials to exhibit excellent performance in a variety of application scenarios, covering fields such as biocarbon fixation, biomedicine, and environmental remediation, and its potential applications extend far beyond these, meeting more complex and diverse needs. The innovation and scalability of this technology provide broader application prospects for materials science, environmental engineering, and life sciences.

[0018] The beneficial effects of this application are reflected in the following aspects: Through an innovatively designed method for constructing photosynthetic microcarriers, and by combining the stability of filamentous fungi with the photosynthetic advantages of microalgae through a symbiotic structure between two different types of bacteria, this approach overcomes the challenges faced by traditional functional living materials, such as poor stability, limited functionality, and insufficient adaptability. This biomimetic design enhances the material's multifunctionality and long-term operability, expanding its application potential in fields such as ecological restoration, resource recycling, and intelligent manufacturing.

[0019] The prepared functional living materials exhibit superior performance in multiple applications, demonstrating high stability, multifunctionality, and excellent adaptability. Employing a symbiotic structure of filamentous fungi and microalgae enhances the material's stability and functionality. The photosynthesis of microalgae and the supporting role of filamentous fungi complement each other, strengthening the material's biocompatibility and long-term operability. By precisely controlling the construction process of photosynthetic microcarriers, the material can self-repair under different environments and achieve functions such as pollution degradation and biological carbon sequestration, significantly improving its application efficiency in ecological restoration and resource recycling. Furthermore, the modular design of the material enables flexible production and promotion capabilities, providing innovative solutions for intelligent manufacturing and environmental protection, and possessing significant academic value and practical application potential.

[0020] The above summary provides a simplified overview of some concepts, which will be further described in detail in the following specific embodiments. The above summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to help determine the scope of the claimed subject matter. The claimed subject matter is not limited to embodiments that address any or all the shortcomings pointed out in the background art.

[0021] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0022] The accompanying drawings, incorporated in and forming part of this specification, illustrate embodiments consistent with this application and, together with the description, serve to explain the principles of this application. To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, those skilled in the art can obtain other drawings based on these drawings without any creative effort. These drawings and textual descriptions are not intended to limit the scope of the concept of this application in any way, but rather to illustrate the concepts of this application to those skilled in the art by referring to specific embodiments.

[0023] Figure 1 This is a schematic diagram of the preparation method of the functional living material in the embodiments of this application; Figure 2 This is a schematic diagram illustrating the preparation of microalgae microspheres in Example 1; Figure 3 This is a schematic diagram of the growth effect of microalgae microspheres in Example 1; Figure 4 This is a schematic diagram of the preparation of filamentous fungal microspheres in Example 1; Figure 5 This is a schematic diagram illustrating the growth effect of filamentous fungal microspheres in Example 1; Figure 6 The shear rate-viscosity curve of the bio-ink shear thinning in Example 1; Figure 7 This is a schematic diagram of the functional living material in Example 1; Figure 8 This is a schematic diagram of the symbiotic relationship between microalgae and fungi in the scaffold in Example 1. Detailed Implementation

[0024] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings. When the following description relates to the drawings, unless otherwise indicated, the same numbers in different drawings denote the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with this application. Rather, they are merely examples of apparatuses and methods consistent with some aspects of this application as detailed in the appended claims.

[0025] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element. It should be further understood that, as used herein, the singular forms "a," "an," and "the" are intended to also include the plural forms, unless the context indicates otherwise. Furthermore, the terms "or," "and / or," "including at least one of the following," etc., as used herein, can be interpreted as inclusive, or mean any one or any combination thereof. Exceptions to this definition only arise when combinations of elements, functions, steps, or operations are inherently mutually exclusive in some manner.

[0026] It should be understood that the specific embodiments described herein are merely illustrative of this application and are not intended to limit the scope of this application.

[0027] Bionics mimics the mechanisms of multicellular symbiotic organisms in nature. Symbionts in nature, such as microalgae and fungi, can perform a variety of ecological and physiological functions through microbial cooperation, providing inspiration for the innovation of functional living materials. Biomimetic design can overcome the limitations of traditional living materials through multicellular synergistic effects, enhanced self-repair capabilities, and improved environmental adaptability. This design not only helps improve the stability and multifunctionality of materials but also opens up new application prospects in fields such as environmental remediation, medical treatment, and energy conversion.

[0028] like Figure 1 As shown, this application innovatively proposes a photosynthetic microcarrier based on the symbiosis of microalgae microspheres and filamentous fungal microspheres, providing an innovative approach to the design of functional living materials. This unique symbiotic structure fully leverages the stability of filamentous fungi and the advantages of microalgae photosynthesis, forming a highly efficient and synergistic composite carrier that significantly enhances the functionality and operability of the material. Microalgae, as the photosynthetic core, can continuously perform photosynthesis under suitable conditions, while filamentous fungi enhance the stability and structural strength of the composite material through their supporting role, extending its long-term operability. This functional living material exhibits excellent biocompatibility and long-term stability, making it widely applicable in fields such as ecological restoration, resource recycling, and intelligent manufacturing, and possessing significant application potential and practical value.

[0029] The present application will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0030] Example 1: Construction of hyperbranched polyethylene glycol diacrylate (HB-PEGDA) and thiolated hyaluronic acid (SH-HA) photosynthetic carriers and their functional living materials (1) Preparation of biopolymer materials HB-PEGDA: Add 13.625g of poly(ethylene glycol) diacrylate (PEGDA) monomer to a 250mL three-necked flask, followed by 62.5mL of butanone solution. Turn on the magnetic stirrer and stir at 300rpm until completely dissolved. Then, add 0.296g of tetraethylthiuram disulfide (DS) and 0.295g of 2,2-azobisisobutyronitrile (AIBN). Purge with argon gas for 1 hour. After oxygen removal, reduce the solution in the three-necked flask to 70°C. o After reaction C for 10 hours, air is introduced to terminate the reaction.

[0031] SH-HA: Dissolve 94.8 mg HA (0.25 mmol) in 10 mL of 0.1 M MES buffer (pH=4.75) at 25 °C for 2.5 h, add 119 mg DTP (0.5 mmol), turn on the magnetic stirrer, and stir at 300 rpm until completely dissolved. Add 96 mg EDCI (0.5 mmol), maintain the pH of the solution at 4.75, react for 5 h, and add 1 M NaOH dropwise to raise the pH of the reaction mixture to 7.0 to stop the reaction. (2) Cultivation of active units Microalgae: Chlorella was cultured in 40 ml of BG11 liquid medium in an artificial incubator at 28°C with a light intensity of 5000-10000 Lux and a light-dark ratio of 12:12 hours for 7-10 days. 5 ml of the medium was then transferred to a new medium to complete one passage.

[0032] Filamentous fungi: Sclerotium truncatum was cultured in PDA solid medium and incubated in a constant temperature and humidity chamber at 25°C and 80% humidity. After 10 days, 1 ml of water was added to rinse the surface of the medium. The seed culture was collected with sterile filter paper and stored in a refrigerator at 4°C.

[0033] (3) Microsphere preparation Weigh 76.8 mg of hyperbranched polyethylene glycol diacrylate (HB-PEGDA) and dissolve it in 960 μL of bacterial culture medium to prepare an 8% (w / v) dispersed phase P1; weigh 38.4... mg of thiolated hyaluronic acid (SH-HA) was dissolved in 1920 μL of bacterial culture medium to prepare a 2% (w / v) dispersed phase P2, and the pH was adjusted to 7.4. Surfactant FE-surf was dissolved in fluorinated oil to prepare a 2% (w / w) continuous phase, designated as oil phase P3. 1000 μL of *Chlorella vulgaris* in its logarithmic growth phase was taken, with the algal concentration adjusted to an OD680 value between 0.3 and 0.5. The algal solution was centrifuged at 4500 rpm for 5 min, and the supernatant was removed. Dispersed phase P1 was added to the remaining substrate, and the mixture was gently shaken to disperse it in P1, yielding a new dispersed phase P4. For filamentous fungi, 100 μL of *Sclerotium sclerotium* bacterial culture activated in seed culture for two days was taken, and the culture was centrifuged at 8000 rpm for 5 min. The supernatant was removed, and the mixture was gently shaken to disperse it in P1, yielding a new dispersed phase P4. Connect the prepared solution to the microfluidic chip via a PET pipe. Adjust the flow rate of dispersed phase P2 to 4 μL / min, the flow rate of continuous phase (oil phase) P3 to 48 μL / min, and the flow rate of dispersed phase P4 to 4 μL / min. Figure 2-5 As shown, the microspheres were collected and placed in a 4°C refrigerator for 3 hours to solidify. After solidification, they were washed out.

[0034] Photosynthetic microcarrier assembly The washed microspheres containing Chlorella and Sclerotium tumefaciens were mixed at a volume ratio of 4:1, and the mixture was gently shaken with a shaker to ensure uniform mixing, thus obtaining a composite photosynthetic microcarrier based on microalgae and fungi.

[0035] (5) Preparation of bio-ink The assembled composite photosynthetic microcarrier is placed in a 10μm sieve, and absorbent paper is placed at the bottom of the sieve to absorb excess water, thus obtaining 3D printing bio-ink. Figure 6 As shown, the photosynthetic microcarrier is a viscoelastic material composed of hydrogel microspheres. When the volume fraction of the microcarrier in the syringe exceeds 58%, a physical jamming effect occurs between the microspheres, causing the material to be in a solid state under low shear force. However, when the external force exceeds a certain level, the particles will slide and exhibit flow characteristics, thus possessing good printability.

[0036] (6) Construction of functional living materials The microcarrier bio-ink obtained in step 5 was filled into a cartridge and printed using an extrusion bioprinter. The printing speed was set to 400 mm / min, and the extrusion pressure was 20 psi. Extrusion was performed using a 21G needle to construct functional living materials, such as… Figure 7 As shown.

[0037] (7) Co-cultivation Weigh 0.102g BG11 and 1.21g PDA and add them to 60mL of pure water. After high temperature and high pressure sterilization, pour them into a petri dish. The ratio of algae culture medium obtained is 1:1. Place the functional scaffold in step (6) into the algae-bacteria mixed culture medium and place the petri dish in a constant temperature and humidity chamber. Set the temperature to 28℃, the light intensity to 5000-10000 Lux, and the light-dark ratio to 12:12 hours in a shaker at 25℃. Figure 8 The symbiotic relationship between algae and bacteria is shown after 24 hours of cultivation.

[0038] Example 2: Construction of hyperbranched polyethylene glycol diacrylate (HB-PEGDA) and thiolated hyaluronic acid thiolated sodium alginate (SH-SA) photosynthetic carriers and their functional living materials The method for preparing functional active materials in this embodiment is the same as that in Example 1. The difference is that the biopolymer material thiolated hyaluronic acid (SH-HA) in step (1) is replaced with thiolated sodium alginate (SH-SA).

[0039] Example 3: Construction of Gel-MA photosynthetic carrier and its functional living material (1) Preparation of biopolymer materials 10 g of type A gelatin was placed in a 250 mL flask, and 100 mL of ultrapure water was added. The mixture was then heated and stirred at 65 °C to dissolve. The temperature was then lowered to 55 °C, and 6 g of methacrylic anhydride was added dropwise. The reaction was continued for 1.5 h. The product was diluted with 100 mL of ultrapure water at 65 °C, centrifuged at 3500 rpm for 10 min, and the supernatant was dialyzed (molecular weight 10000) for one week. The pH of the product solution was adjusted to 6.5-7, and then freeze-dried.

[0040] (2) Cultivation of active units The same as step (2) in Example 1.

[0041] (3) Microsphere preparation The surfactant (FE-surf) was dissolved in fluorinated oil to prepare a 2% (w / v) solution, denoted as the continuous phase P1 (oil phase). 80 mg of double-bonded gelatin (Gel-MA) was dissolved in 1000 μL of bacterial culture medium, and 5 mg of photoinitiator LAP was added to prepare an 8% (w / v) solution, denoted as the dispersed phase P2. Microalgae and fungi were dispersed evenly in P2 to obtain a new dispersed phase P3. The prepared solution was connected to a microfluidic chip through a PET tube. The flow rate of oil phase P1 was adjusted to 40 μL / min, and the flow rate of dispersed phase P3 was adjusted to 4 μL / min. After collection, the solution was cured under blue light for 300 s, and the microspheres were washed out.

[0042] (4) — (7) The steps (4) to (7) are the same as in Example 1.

[0043] Example 4: Construction of double-bonded F127 (F127-DA) photosynthetic carrier and its functional living material (1) Preparation of biopolymer materials Weigh 10g of F127 and place it in a 250mL flask. Place the flask in an 80℃ vacuum drying oven for 5 hours, then cool to room temperature. Add 0.119g (0.99mmol) of chloroform and stir in an ice bath to dissolve. After F127 is completely dissolved, add 0.7968g (0.249mmol) of triethylamine. Add 0.11g of acryloyl chloride to 20mL of chloroform dropwise through a constant pressure burette. React at room temperature for 48 hours. After the reaction is complete, filter off the precipitate. After rotary evaporation, add excess diethyl ether and filter again. Collect the white precipitate on the filter paper, dissolve it again in a small amount of chloroform, add diethyl ether, and filter again. Dry the final white precipitate at room temperature.

[0044] (2) Cultivation of active units The same as step (2) in Example 1.

[0045] (3) Microsphere preparation The surfactant (FE-surf) was dissolved in fluorinated oil to prepare a 2% (w / v) continuous phase, denoted as P1 (oil phase). 150 mg of double bond F127 (F127-DA) was weighed and dissolved in 1000 μL of bacterial culture medium, and 5 mg of photoinitiator LAP was added to prepare a 15% (w / v) dispersed phase, denoted as P2. Microalgae and fungi were dispersed evenly in P2 to obtain a new dispersed phase, P3. The prepared solution was connected to a microfluidic chip through a PET tube. The flow rate of oil phase P1 was adjusted to 20 μL / min, and the flow rate of dispersed phase P3 was adjusted to 2 μL / min. The algae-coated gel microspheres were collected after 1 h, cured with blue light for 300 s, and the microspheres were washed out.

[0046] (4) — (7) The steps (4) to (7) are the same as in Example 1.

[0047] Example 5: Construction of double-bonded sodium alginate (SA-MA) photosynthetic carrier and its functional living material Preparation of biopolymer materials 1 g of sodium alginate and 7.5 mL of methacrylic anhydride were added to 100 mL of deionized water and mixed thoroughly to obtain a reaction solution. The pH of the reaction solution was adjusted to 8 with 5 mol / L NaOH solution, and then the reaction was stirred at 0 °C for 24 h (600 rpm / min). After the reaction was completed, the polymer product was washed with ethanol, dialyzed for 3 days, and freeze-dried to obtain double-bonded sodium alginate (SA-MA). Cultivation of active units The same as step (2) in Example 1.

[0048] (3) Microsphere preparation The surfactant (FE-surf) was dissolved in fluorinated oil to prepare a 2% (w / v) continuous phase, denoted as P1 (oil phase). 30 mg of double bond F127 (F127-DA) was weighed and dissolved in 1000 μL of bacterial culture medium, and 5 mg of photoinitiator LAP was added to prepare a 15% (w / v) dispersed phase, denoted as P2. Microalgae and fungi were dispersed evenly in P2 to obtain a new dispersed phase, P3. The prepared solution was connected to a microfluidic chip through a PET tube. The flow rate of oil phase P1 was adjusted to 20 μL / min, and the flow rate of dispersed phase P3 was adjusted to 2 μL / min. The algae-coated gel microspheres were collected after 1 h, cured with blue light for 300 s, and the microspheres were washed out.

[0049] (4) — (7) The steps (4) to (7) are the same as in Example 1.

[0050] The sequence numbers of the embodiments in this application are for descriptive purposes only and do not represent the superiority or inferiority of the embodiments.

[0051] In this application, the same or similar terms, concepts, technical solutions and / or application scenario descriptions are generally described in detail only when they appear for the first time. When they appear again, they are generally not repeated for the sake of brevity. When understanding the technical solutions and other contents of this application, the same or similar terms, concepts, technical solutions and / or application scenario descriptions that are not described in detail later can be referred to their previous relevant detailed descriptions.

[0052] In this application, the descriptions of the various embodiments have different focuses. For parts that are not described in detail or recorded in a certain embodiment, please refer to the relevant descriptions of other embodiments.

[0053] The technical features of the present application can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of the present application.

[0054] The above are merely preferred embodiments of this application and do not limit the patent scope of this application. Any equivalent structural or procedural transformations made using the content of this application's specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of this application.

Claims

1. A method for preparing functional living materials using photosynthetic microcarriers, characterized in that, The method includes the following steps: A first microbial microsphere and a second microbial microsphere were prepared using a droplet microfluidic method, wherein the first microbial microsphere encapsulated microalgae and the second microbial microsphere encapsulated filamentous fungi. The first microbial microspheres and the second microbial microspheres are mixed to form a composite microcarrier; The composite microcarrier is constructed into a functional living material with a specific shape by 3D printing and then placed in a culture environment for cultivation, so that the microalgae and the filamentous fungi form a symbiotic system.

2. The method according to claim 1, characterized in that, The first and second microbial microspheres further include biopolymer materials, which include one or more of the following: thiolated hyaluronic acid (SH-HA), hyperbranched polyethylene glycol diacrylate (HB-PEGDA), thiolated sodium alginate (SH-SA), double-bonded gelatin (Gel-MA), and double-bonded F127 (F127-DA).

3. The method according to claim 1 or 2, characterized in that, The microalgae include one or more of Spirulina, Chlorella, cyanobacteria, genetically modified Spirulina, genetically modified Chlorella, and genetically modified cyanobacteria.

4. The method according to claim 1 or 2, characterized in that, The filamentous fungi include Aspergillus niger, Polyporus genus, Sclerotium sclerotiorum, genetically engineered Aspergillus niger or genetically engineered Polyporus genus.

5. The method according to claim 2, characterized in that, The droplet microfluidic method includes: uniformly dispersing the microalgae or the filamentous fungi in the biopolymer material as a dispersed phase, dispensing the dispersed phase into microdroplets using continuous phase shearing, and then cross-linking and solidifying to form the first microbial microsphere and the second microbial microsphere.

6. The method according to claim 5, characterized in that, The cross-linking and curing process includes any one of the following cross-linking methods: Michael addition reaction, photocross-linking, esterification reaction, and ionic cross-linking. The diameters of the first microbial microspheres and the second microbial microspheres are 100-300 μm, respectively.

7. The method according to claim 1 or 2, characterized in that, The first microbial microspheres and the second microbial microspheres are mixed at a volume ratio of 4:1 to form the composite microcarrier.

8. The method according to claim 1 or 2, characterized in that, The culture environment conditions are as follows: a culture medium prepared by mixing BG11 and PDA in a 2:1 ratio, a temperature of 28-30℃, a light intensity of 5000-10000 Lux, a light-dark ratio of 12:12 hours, and a humidity of 50-60% RH.

9. The method according to claim 1 or 2, characterized in that, The 3D printing method involves filling the composite microcarrier into a printing syringe and printing a scaffold model with a specific shape using a 3D printing device; wherein the 3D printing device includes any one of an extrusion printing device, an inkjet printing device, or a photopolymerization printing device.

10. A functional living material prepared using photosynthetic microcarriers, characterized in that, The functional living material is prepared by the method according to any one of claims 1-9. The microalgae in the functional living material produce oxygen and organic matter through photosynthesis, and the filamentous fungi secrete hyphae using carbon sources in the external environment and extend and grow along the preset printing path.

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