An environmentally friendly bio-growth self-assembly method of room temperature phosphorescent mycelium material

Abstract

The application discloses a kind of environmental friendly room temperature phosphorescence mycelium material biological growth self-assembly method, belong to biotechnology field.In the process of fungal growth, sulfonic acid group with different pi conjugated structure is self-assembled into mycelium by biological growth, thereby inhibiting the non-radiative decay of triplet exciton, realizing efficient room temperature phosphorescence emission.And by changing the conjugation degree of chromophore, full color adjustable afterglow from blue to red is realized.Under the above technical scheme, the room temperature phosphorescence mycelium material of the application is self-assembled by biological growth, which simplifies the production process and reduces the generation of carbon footprint in the process, not only can be completely degraded in nature, but also can improve soil fertility to promote plant growth.The method is helpful to develop environmentally friendly room temperature phosphorescence material and expand the application of fungi in the field of functional optics.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material. Background Technology

[0002] Room-temperature phosphorescent (RTP) materials have attracted much attention due to their long lifetime, large Stokes shift, and high luminous efficiency. With these unique photophysical properties, RTP materials have broad application prospects in many fields, including information encryption, sensors, bioimaging, and 3D printing. Inorganic, organic, and natural biomass-based RTP materials have also been extensively studied. However, due to limitations in resources, cost, and preparation methods, the development of truly green, economical, and mass-producible organic RTP materials is urgently needed.

[0003] Bioluminescence is a common phenomenon in nature, with various organisms such as fungi, bacteria, and even fireflies possessing efficient, environmentally friendly, and fully biodegradable luminescence capabilities. These natural systems provide valuable inspiration and a biomimetic basis for the design and construction of sustainable artificial luminescent materials.

[0004] Fungal resources, as a type of biomaterial, are diverse, grow rapidly, and have a near-unlimited reproductive capacity. The supply of raw materials is almost unrestricted, and the production process does not require complex technological steps or toxic chemical reagents. From cultivation to molding, they maintain natural and safe characteristics, making them a truly sustainable and environmentally friendly emerging luminescent material resource. Summary of the Invention

[0005] Inspired by the phenomenon of bioluminescence, this invention proposes a biological growth self-assembly method, which directly obtains environmentally friendly room temperature phosphorescent mycelial materials through the biological growth self-assembly process of fungi.

[0006] This invention provides a bio-growth self-assembly method for environmentally friendly room-temperature phosphorescent mycelial materials. Through the bio-growth self-assembly process, environmentally friendly functional room-temperature phosphorescent mycelial materials are obtained, laying an important foundation for expanding the application of fungi in the field of optics.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solution: The present invention discloses a method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material, which includes the following steps: (1) The fungus *Porphyromonas villiformis* Trametes pubescens The (MTP) strain was inoculated onto a solid culture medium for activation and then incubated at 28 °C for 5 days. (2) The activated strain was inoculated into malt extract liquid culture medium and placed in a constant temperature shaking culture at 28 ℃ and 150 r / min for 5 days to prepare seed fermentation broth; (3) Use an internal homogenizer to stir the seed fermentation liquid at 5000 r / min for 1 min to form a uniform mycelial seed liquid; (4) Inoculate 10 mL of seed culture into a liquid medium containing 10 mg of sulfonic acid chromophores [(SAC): including amino-1,4-benzenedisulfonic acid (BZS), 4,4'-biphenyldisulfonic acid (BPS), 1,5-naphthalenedisulfonic acid (NLS), 4-sulfonic acid-1,8-naphthalenedicarboxylic anhydride (NAS) and 1-pyrenesulfonic acid (PYS)] and 100 mL of malt extract powder, and place it in the dark, at 28 ℃, 50% relative humidity, and 150 r / min constant temperature and humidity shaking culture for 7 days to obtain mycelium; (5) The mycelium was thoroughly rinsed with deionized water to remove the culture medium residue, and then the solid products of room temperature phosphorescent mycelium (SAC-MTP: BZS-MTP, BPS-MTP, NLS-MTP, NAS-MTP and PYS-MTP) were obtained by freeze drying.

[0008] This invention also discloses the application of room temperature phosphorescent mycelium materials in information encryption and luminescent decoration.

[0009] This invention also discloses the biodegradability of room temperature phosphorescent mycelium materials.

[0010] During fungal growth, SACs with different π-conjugated structures self-assemble into the mycelium through biological growth, thereby suppressing the non-radiative decay of triplet excitons and achieving efficient room-temperature phosphorescence emission. Furthermore, by changing the degree of conjugation of the chromophores, a full-color tunable afterglow ranging from blue to red is achieved.

[0011] Compared with the prior art, the beneficial effects of the present invention are: This invention discloses a method for the bio-growth and self-assembly of room-temperature phosphorescent mycelial materials. Under the same conditions, the prepared room-temperature phosphorescent mycelial materials, compared with other common room-temperature phosphorescent materials, exhibit complete biodegradability and minimal environmental impact. This method simplifies the preparation process of room-temperature phosphorescent materials, reduces production costs, minimizes environmental impact, and aligns with green environmental protection principles. The prepared room-temperature phosphorescent mycelial materials achieve full-color tunable long-lifetime afterglow emission from blue to cyan to green to yellow to red. The materials of this invention show promising applications in information encryption, anti-counterfeiting, and luminescent decoration. Attached Figure Description

[0012] Figure 1Figure 1 shows the control group of MTP and SAC-MTP. Figure 2 shows stereomicroscopic and scanning electron microscope images of MTP (control group) and SAC-MTP (SAC concentration: 0.1 mg / mL) cultured; Figure 3 shows the diameter of hyphal balls and hyphae of MTP and SAC-MTP (SAC concentration: 0.1 mg / mL); Figure 4 shows the content of extracellular polysaccharides secreted by MTP and BPS-MTP (different concentrations of BPS); Figure 5 shows the biomass of MTP and SAC-MTP (SAC concentration: 0.1 mg / mL). Figure 2 This section compares the steady-state photoluminescence, delayed emission, and phosphorescence lifetime performance of SAC-MTP. In this section, a represents the steady-state photoluminescence (dashed line) and delayed emission (solid line) spectra, corresponding to BZS-MTP (excitation wavelength: 310 nm), BPS-MTP (excitation wavelength: 254 nm), NLS-MTP (excitation wavelength: 310 nm), NAS-MTP (excitation wavelength: 365 nm), and PYS-MTP (excitation wavelength: 365 nm), respectively; delay time: 1 ms; b represents the afterglow image of SAC-MTP; c represents the International Commission on Illumination (CIE) coordinates corresponding to the afterglow emission of SAC-MTP; and d represents the time-resolved decay curve of SAC-MTP. Figure 3 This section describes the performance and applications of SAC-MTP materials. Specifically: a) large-area manufacturing and recyclable SAC-MTP; b) an example of SAC-MTP used as ink for anti-counterfeiting applications; c) mushroom-shaped paper made with SAC-MTP, capable of repeated writing with water as ink; d) various shapes of luminescent decorations made with SAC-MTP; and e) the phosphorescence intensity of BZS-MTP, BPS-MTP, NAS-MTP, and PYS-MTP over 35 days. Figure 4 The biodegradability and environmental impact of SAC-MTP are shown in Figure a. a represents the biodegradability of SAC-MTP (right), wood pulp paper (W paper) (left), and polyvinyl chloride (PVC) film (middle) in a soil environment; b represents the growth of Chinese cabbage; c represents the biodegradability of BZS-MTP, a-CDs / BA, and Cd3Al2Ga3O4. 12 :Ce 3+ Environmental impact. Detailed Implementation

[0013] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0014] This invention provides the application of a strain in the preparation of environmentally friendly room-temperature phosphorescent mycelial materials.

[0015] In some embodiments, the strains include not only *Fomitopsis pinicola*. Trametes pubescens (MTP), also includes the genus *Pterocarpus*. Trametes Ganoderma genus Ganoderma Pleurotus Pleurotus These fungi, whose strains and cultures are all white, belong to the white decay fungi family and can also be used to prepare environmentally friendly room-temperature phosphorescent mycelial materials. These materials are completely biodegradable and have minimal environmental impact. They also show promising applications in information encryption, anti-counterfeiting, and luminescent decoration.

[0016] Example 1: Growth of SAC-MTP (1) The fungus *Porphyromonas villiformis* Trametes pubescens The strain was inoculated onto solid culture medium (300 g / L peeled potato, 20 g / L glucose, 20 g / L agar, 1 g / L potassium dihydrogen phosphate, pH 5, 1×10⁻⁶ ppm). 5 Activate on a sterile surface (sterilize for 30 min), and incubate at 28 ℃ for 5 days; (2) The activated strain was inoculated into malt extract liquid medium (malt extract 20 g / L, glucose 10 g / L, potassium dihydrogen phosphate 3 g / L, vitamin B1 0.01 g / L, pH 5, 1×10⁻⁶). 5 The mixture was sterilized in Pa for 30 min and then placed in a constant temperature shaking culture at 28 ℃ and 150 r / min for 5 days to prepare the seed fermentation broth. (3) Use an internal homogenizer to stir the seed fermentation liquid at 5000 r / min for 1 min to form a uniform mycelial seed liquid; (4) Inoculate 10 mL of seed culture into a liquid medium containing 10 mg of sulfonic acid chromophores [SAC: including amino-1,4-benzenedisulfonic acid (BZS), 4,4'-biphenyldisulfonic acid (BPS), 1,5-naphthalenedisulfonic acid (NLS), 4-sulfonic acid-1,8-naphthalenedicarboxylic anhydride (NAS) and 1-pyrenesulfonic acid (PYS)] and 100 mL of malt extract powder, and place it in the dark, at 28 ℃, 50% relative humidity, and 150 r / min constant temperature and humidity shaking culture for 7 days to obtain mycelium; During liquid culture, uniform SAC-MTP and MTP mycelial balls were generated, and no obvious morphological differences were observed. Figure 1 a). The average diameter of SAC-MTP mycelial balls is approximately 0.15 cm ( Figure 1 b). Inside the hyphal ball, the interweaving of hyphae forms a complex three-dimensional network structure, with hyphae diameters of approximately 2 μm ( Figure 1 b). Sulfur in SAC promotes fungal biomass growth and the production of extracellular polysaccharides. In a flowing liquid environment, the secretion of extracellular polysaccharides enables hyphae to adhere to each other, thereby promoting the formation of hyphal balls. Figure 1c). After cultivation, the final SAC-MTP biomass obtained was 0.36-1.39 g ( Figure 1 d). The mycelial balls and highly water-soluble SAC can synergistically enhance each other, enabling the room-temperature phosphorescent mycelium to not only efficiently carry out biological growth and self-assembly, but also to ensure the normal growth and metabolic activities of the host fungus.

[0017] Example 2: Photophysical characterization of SAC-MTP The obtained mycelium was thoroughly rinsed with deionized water to remove culture medium residue, and then freeze-dried to obtain room temperature phosphorescent mycelium (SAC-MTP: BZS-MTP, BPS-MTP, NLS-MTP, NAS-MTP and PYS-MTP) solid products.

[0018] Photophysical properties of SAC-MTP: Steady-state photoluminescence, delayed emission, and phosphorescence lifetime of SAC-MTP were measured using an Edinburgh FLS 980 fluorescence spectrometer equipped with a xenon lamp, LED array, and microsecond lamp. Afterglow photos were taken using an iPhone 14 at room temperature.

[0019] like Figure 2 As shown in the steady-state photoluminescence spectra, the fluorescence peaks of BZS-MTP, BPS-MTP, NLS-MTP, NAS-MTP, and PYS-MTP are located at 476, 402, 487, 471, and 423 nm, respectively. All SAC-MTP materials exhibit blue or blue-violet fluorescence under ultraviolet excitation. Figure 2 b). After removing ultraviolet light, the multicolored afterglow emission (blue, cyan, green, yellow, and red) can be clearly observed. Figure 2 b). The delayed spectra of BZS-MTP, BPS-MTP, NLS-MTP, NAS-MTP, and PYS-MTP show emission peaks at 466, 490, 525, 601, and 635 nm, respectively. Figure 2 a), the corresponding CIE coordinates are (0.18, 0.25), (0.23, 0.38), (0.36, 0.52), (0.47, 0.46) and (0.39, 0.37) respectively. Figure 2 c), consistent with the color of the afterglow as observed with the naked eye ( Figure 2 b). Time-resolved decay curves show that the lifetimes of BZS-MTP, BPS-MTP, NLS-MTP, NAS-MTP, and PYS-MTP are 440, 910, 890, 340, and 200 ms, respectively. Figure 2 d).

[0020] Example 3: Application Potential of SAC-MTP Applications of SAC-MTP: To transform SAC-MTP into luminescent decorations or mushroom-shaped paper, washed SAC-MTP is stirred at 5000 r / min for 2 min using an internal cutting homogenizer. The completely homogenized SAC-MTP slurry is then freeze-dried in a suitable silicone mold to produce luminescent decorations of various shapes. Mushroom-shaped paper is produced by vacuum membrane filtration and freeze-drying of the SAC-MTP slurry.

[0021] Stability of SAC-MTP: The phosphorescence intensity of SAC-MTP was measured periodically using a fluorescence spectrometer within 35 days.

[0022] The cleaned SAC-MTP is uniformly mixed into a slurry, which is then processed into various materials using a scalable molding process. This mushroom-shaped paper exhibits excellent recyclability. After use, it can be cut, ground, and redispersed in warm water to form a homogeneous slurry, which can then be reused as a raw material for manufacturing room-temperature phosphorescent products. Figure 3 a). Amorphous SAC-MTP materials show great potential in information encryption, anti-counterfeiting, and luminescent decoration. SAC-MTP slurry has strong adhesion and can be used directly as ink for writing and drawing. Figure 3 b). SAC-MTP slurry is processed into mushroom paper through vacuum membrane filtration and freeze-drying. Since water inhibits room temperature phosphorescence due to the disruption of hydrogen bonds, water can be used as ink to repeatedly write on the surface of the mushroom paper. Figure 3 c). Various shapes of models can also be created by injecting SAC-MTP slurry into silicone molds, including flowers, cats, hearts, mushrooms, and carrots, etc. Figure 3 d). At room temperature, the phosphorescence intensity of SAC-MTP showed virtually no significant attenuation after more than 35 days. Figure 3 e), thus it can be seen that SAC-MTP materials have reliable long-term stability in room temperature applications.

[0023] Example 4: Biodegradability of SAC-MTP Biodegradability of SAC-MTP: The prepared mushroom paper was buried in soil at a depth of 3 cm along with two common RTP materials [wood pulp paper (W paper) and polyvinyl chloride (PVC) film]. Morphological changes in the paper were monitored regularly to assess the degradation process. 6.25% SAC-MTP was added to the soil, and six Chinese cabbage seeds were sown; seedling growth was systematically monitored.

[0024] Life cycle assessment of SAC-MTP: A life cycle assessment model was established using SimaPro 10.1 software and the Ecoinvent 3.10 database, and the impact was assessed using the ReCiPe 2016 Midpoint (H) V1.08 / World (2010) H method. Taking 1 kg of BZS-MTP mycelial material as an example, it was compared with organic phosphorescent materials (a-CDs / BA) and inorganic phosphorescent materials (Cd3Al2Ga3O). 12 :Ce 3+ (Compare)

[0025] By day 35, SAC-MTP was completely degraded. Figure 4 a right), W paper ( Figure 4 (left) is broken, PVC film ( Figure 4 (a) It retains its original form. This indicates that SAC-MTP can be effectively decomposed by the natural environment at the end of its lifespan. Furthermore, SAC-MTP can effectively improve soil fertility and significantly promote plant growth during its degradation process, demonstrating excellent ecological safety. Figure 4 b). With organic a-CDs / BA and inorganic Cd3Al2Ga3O 12 :Ce 3+ Compared to the two reference phosphorescent materials, BZS-MTP has a significantly lower environmental impact across all categories. Figure 4 c).

[0026] In this invention, an environmentally friendly room-temperature phosphorescent mycelium material was obtained through the self-assembly process of fungal biological growth. This method helps to develop environmentally friendly room-temperature phosphorescent materials and expands the application of fungi in the field of functional optics.

Claims

1. A method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material, characterized in that, The method includes the following steps: (1) The fungus *Porphyromonas villiformis* Trametes pubescens The strain was inoculated onto a solid culture medium for activation and then incubated at 28 ℃ for 5 days. (2) The activated strain was inoculated into malt extract liquid culture medium and placed in a constant temperature shaking culture at 28 ℃ and 150 r / min for 5 days to obtain seed fermentation broth; (3) Use an internal homogenizer to stir the seed fermentation liquid at 5000 r / min for 1 min to form a uniform mycelial seed liquid; (4) Inoculate 10 mL of seed liquid into a liquid culture medium containing 10 mg of sulfonic acid chromophore and 100 mL of malt extract powder, and place it in the dark, at 28 ℃, 50% relative humidity, and 150 r / min constant temperature and humidity shaking culture for 7 days to obtain mycelium; (5) The mycelium was thoroughly rinsed with deionized water to remove the culture medium residue, and then freeze-dried to obtain room temperature phosphorescent mycelium solid product.

2. The method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material according to claim 1, characterized in that, The sulfonic acid chromophore is one of amino-1,4-benzenedisulfonic acid, 4,4'-biphenyldisulfonic acid, 1,5-naphthalenedisulfonic acid, 4-sulfonic acid-1,8-naphthalenedicarboxylic anhydride, and 1-pyrenesulfonic acid.

3. The method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material according to claim 1, characterized in that, Solid culture medium: 300 g / L peeled potato, 20 g / L glucose, 20 g / L agar, 1 g / L potassium dihydrogen phosphate, pH 5, 1×10⁻⁶ 5 Sterilize for 30 minutes using Pa.

4. The method for the bio-growth and self-assembly of an environmentally friendly room-temperature phosphorescent mycelium material according to claim 1, characterized in that, Malt extract liquid culture medium: 20 g / L malt extract, 10 g / L glucose, 3 g / L potassium dihydrogen phosphate, 0.01 g / L vitamin B1, pH 5, 1×10⁻⁶ 5 Sterilize for 30 minutes using Pa.

5. A room-temperature phosphorescent mycelium material assembled by a bio-growth self-assembly method according to any one of claims 1-4.

6. The application of the room temperature phosphorescent mycelium material of claim 5 in information encryption, anti-counterfeiting and luminescent decoration.

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