Edible mushroom mycelium particle stabilized lycopene pickering emulsion and preparation method thereof

Abstract

The application discloses edible mushroom mycelium particle stable lycopene Pickering emulsion and a preparation method thereof, and relates to the field of food technology. The method uses edible mushroom mycelium whole biomass particles retaining protein, polysaccharide, dietary fiber, chitin and cell wall structure as stabilizers, and obtains a particle dispersion liquid by sequentially performing hydration, low-speed pre-dispersion, high-speed shearing and cooling. Lycopene is prepared into an oil phase, the particle dispersion liquid is subjected to vacuum pulse rehydration and a small amount of oil phase pre-wetting, the remaining oil phase is added in batches and alternately subjected to short-time shearing and light-shielded standing, and finally, terminal whole-grain and low-shear interface rearrangement treatment are performed to obtain an O / W type Pickering emulsion. The method does not need to extract protein or polysaccharide from the mycelium and perform chemical modification, can improve the adsorption and coverage of the particles at the oil-water interface, and improves the uniformity of emulsion droplets, the storage stability, and the lycopene encapsulation and retention effect.

Description

Technical Field

[0001] This invention relates to the field of food technology, and in particular to a lycopene Pickering emulsion stabilized in edible fungi mycelium particles and its preparation method. Background Technology

[0002] Lycopene is a fat-soluble carotenoid with antioxidant activity and nutritional functions, and can be used in functional foods, nutritional supplements, and natural coloring systems. However, lycopene has poor water solubility, and its molecule contains multiple conjugated double bonds, making it prone to oxidative degradation or isomerization under light, oxygen, temperature changes, and during food processing. When lycopene powder, crystals, or lycopene-containing oil phases are directly added to aqueous food products, it is usually difficult to form a uniform and stable dispersion system, easily leading to problems such as oil droplet aggregation, layering, oil separation, uneven color, and decreased lycopene retention during storage. Existing technologies typically use small-molecule surfactants, proteins, modified starches, colloidal polysaccharides, or protein-polysaccharide complexes as emulsifying stabilizers, and prepare lycopene emulsions through high-speed shearing and high-pressure homogenization. However, some systems suffer from problems such as large emulsifier usage, reliance on chemically modified materials, complex preparation processes, and low overall utilization of natural raw materials.

[0003] Pickering emulsions utilize solid particles adsorbed at the oil-water interface to form a particle-stabilized layer, reducing the possibility of direct contact and aggregation between adjacent oil droplets. Existing food-grade pickering stabilizers mainly include starch particles, cellulose particles, protein particles, polysaccharide particles, and protein-polysaccharide composite particles. These particle systems typically require extraction, purification, modification, compounding, or particle size control to obtain suitable dispersibility and interfacial stability. King oyster mushroom mycelium contains proteins, polysaccharides, dietary fiber, chitin, and cell wall structures, making it a potential food-grade pickering stabilizer. However, its irregular particle morphology, porosity, and fibrous nature make it prone to aggregation during direct hydration, and air may remain inside the particles. Under conditions of single-phase addition of the oil phase and continuous high-speed shearing, the rate of oil droplet formation is difficult to match with the rate of particle migration, adsorption, and rearrangement at the oil-water interface, easily leading to incomplete particle coverage on the oil droplet surface, uneven droplet size distribution, and insufficient emulsion storage stability. Therefore, how to improve the particle dispersibility and oil-water interface adsorption capacity of Pleurotus eryngii mycelium while preserving its natural multi-component structure, and form a Pickering emulsion with uniform droplets, stable interface, and beneficial protection of lycopene, is a technical problem that needs to be solved in this field. Summary of the Invention

[0004] This application provides a method for preparing a lycopene Pickering emulsion stabilized in edible fungi mycelium particles, comprising the following steps: S1. Take the whole biomass of edible fungi mycelium, and after washing, dehydration, drying, crushing and sieving, obtain granules of whole biomass of edible fungi mycelium; S2. Add the edible fungus mycelium biomass particles to water and stir to hydrate them, to obtain an initial biomass particle dispersion; S3. The initial whole biomass particle dispersion is subjected to low-speed pre-dispersion, high-speed shear dispersion and ice bath cooling treatment in sequence to obtain the whole biomass particle dispersion. S4. Add lycopene to edible oil and stir to dissolve or disperse it under light-protected conditions to obtain the lycopene oil phase; S5. The whole biomass particle dispersion obtained in step S3 is subjected to vacuum pulse rehydration treatment. Part of the lycopene oil phase obtained in step S4 is added to the whole biomass particle dispersion that has been vacuum pulse rehydration treatment for pre-wetting treatment, so as to obtain the edible fungus mycelium whole biomass particle dispersion with pre-activated interface. S6. Add the remaining lycopene oil phase obtained in step S4 to the pre-activated edible fungus mycelium whole biomass particle dispersion obtained in step S5 in batches. After each batch of lycopene oil phase is added, short-time shearing and intermittent standing are performed in sequence. After all the lycopene oil phase is added, terminal granulation is performed to obtain lycopene Pickering emulsion. S7. The lycopene Pickering emulsion is subjected to low-shear interface rearrangement treatment to obtain a lycopene Pickering emulsion with stable whole biomass particles of edible fungi mycelium.

[0005] It should be noted that step S1, through washing, drying, pulverizing, and sieving, forms whole biomass particles with a porous fibrous structure while retaining the natural complex state of proteins, polysaccharides, dietary fiber, chitin, and cell wall structure in the mycelium; step S2 allows the aqueous phase to gradually enter the pores of the particles and the gaps between the mycelial bundles, promoting water absorption and swelling of the particles and reducing aggregation; the low-speed pre-dispersion in step S3 ensures uniform distribution of hydrated particles and establishes stable shear transfer conditions, while high-speed shearing further dissociates the mycelial bundles and particle aggregates, exposing natural hydrophilic and hydrophobic sites; ice bath cooling dissipates shear heat in a timely manner and provides conditions for continued hydration and structural relaxation of the particles, thereby obtaining a uniformly dispersed whole biomass particle dispersion that retains its natural complex structure; step S4 uniformly loads lycopene into edible oil under light-protected conditions; and step S5 involves vacuum pulse rehydration. Residual air in the particle pores is expelled and the aqueous phase is further penetrated into the particle interior. Then, a small amount of lycopene oil phase is used to pre-wet the particle surface, inducing the natural amphiphilic component to form an orientation state suitable for adsorption at the oil-water interface. In step S6, the remaining oil phase is added in batches, and short-term shearing and intermittent standing are performed between batches to gradually match the formation of oil droplets with the migration, adsorption, and rearrangement of particles to the oil-water interface. Then, the final particle sizing makes the size and distribution of emulsion droplets tend to be uniform. In step S7, low-shear interface rearrangement promotes the further arrangement, interlocking, and locking of adsorbed particles, and finally forms a relatively continuous particle interface covering structure on the surface of lycopene oil droplets, thereby improving the spatial barrier effect between oil droplets, thus improving the overall emulsification uniformity, physical stability, and encapsulation and protection effect of lycopene Pickering emulsion.

[0006] Furthermore, in step S1, the edible fungus mycelium is king oyster mushroom mycelium.

[0007] It should be noted that the proteins, polysaccharides, dietary fiber, chitin, and cell wall structures in the whole biomass of Pleurotus eryngii mycelium coexist in a natural complex state. After hydration and physical dispersion, they can form particles that have both aqueous dispersibility and oil-water interface adsorption capacity, providing an interfacial stability basis for the subsequent construction of a stable lycopene O / W type Pickering emulsion.

[0008] Furthermore, in step S2, the mass concentration of edible fungi mycelium whole biomass particles in the initial whole biomass particle dispersion is 3-7 g / 100 mL, and the stirring hydration time is 60 min.

[0009] It should be noted that the combination of a particle mass concentration of 3–7 g / 100 mL and 60 min of stirring and hydration allows the aqueous phase to fully enter the pores and interfiber spaces of the mycelial particles, and maintains the appropriate fluidity of the dispersion while ensuring the number of particles and the viscoelasticity of the continuous phase, thereby obtaining an initial whole biomass particle dispersion with uniform hydration and low degree of agglomeration.

[0010] Furthermore, step S3 includes the following steps: S31. The initial whole biomass particle dispersion is pre-dispersed at 600 r / min for 10 min to obtain a pre-dispersed solution; S32. The pre-dispersed liquid is subjected to high-speed shearing at 16000 r / min for 5 min to obtain a sheared dispersion; S33. Cool the shear dispersion in an ice bath for 3 minutes; Steps S32 and S33 constitute a shear-cooling cycle, which is performed twice to obtain the whole biomass particle dispersion.

[0011] It should be noted that in step S31, pre-dispersion at 600 r / min for 10 min is performed to evenly spread particles with different degrees of hydration, adjust the particle spacing, and establish uniform shear transfer conditions. Based on this, in step S32, high-speed shearing at 16000 r / min for 5 min is performed to dissociate the hyphae and particle aggregates using shear force and turbulence, making the particle size more uniform and forming new particle surfaces and pore channels, promoting the appropriate exposure of hydrophilic and hydrophobic sites of natural components such as proteins, polysaccharides, dietary fiber, and chitin. Step S33 is then followed by ice bath cooling for 3 minutes. The process not only removes the heat generated by shearing in a timely manner, but also allows moisture to continue to enter the newly exposed surfaces and pores after shearing, providing time for the fiber structure to relax and the particle hydration state to rebalance. This allows the particles formed by the first shearing to enter the second shearing with a more stable structure. The second shearing further depolymerizes the fully rehydrated residual hyphae bundles and aggregated regions. Subsequent ice bath cooling stabilizes the resulting particle structure, ensuring that particle depolymerization, surface interface activation, and the natural composite structure are coordinated. Without extracting single components or performing chemical modifications, the mycelial biomass is transformed into a particle dispersion system with a relatively uniform particle size distribution, low degree of aggregation, and suitable oil-water interface adsorption capacity, providing a stable foundation for subsequent vacuum pulse rehydration, oil phase prewetting, and particle interface film formation.

[0012] Furthermore, in step S4, the edible oil is sunflower seed oil, and the lycopene content in the lycopene oil phase is 0.10% by mass; the lycopene oil phase is prepared by stirring at 500 r / min for 30 min under light-protected conditions.

[0013] It should be noted that sunflower seed oil provides a compatible lipid carrier for lycopene. A 0.10% mass percentage content balances lycopene loading with uniform oil phase dispersion. Lycopene and sunflower seed oil are thoroughly mixed under light-protected conditions with stirring at 500 r / min for 30 min, ensuring a stable dissolved or dispersed state of lycopene in the oil phase. The resulting oil phase is used for subsequent pre-wetting and emulsification treatments. The actual solubility of lycopene in the oil phase can be confirmed through centrifugation, microscopic observation, or quantitative analysis.

[0014] Furthermore, step S5 includes the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08MPa and maintain for 3min, restore to normal pressure and let stand for 3min. Repeat the evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase obtained in step S4 and add it to the whole biomass particle dispersion after vacuum pulse rehydration treatment. Stir at 800 r / min for 8 min and then let stand for 15 min in the dark to obtain the pre-activated edible fungus mycelium whole biomass particle dispersion.

[0015] It should be noted that in step S51, the residual gas in the pores and interfiber gaps of the mycelial biomass particles expands and is expelled by maintaining a pressure of −0.08 MPa for 3 min. After restoring to normal pressure and standing for 3 min, the pressure recovery is used to push the aqueous phase further into the particle interior. This process is repeated 3 times to complete the degassing, water infiltration, and structural relaxation in stages, so that the particles form a relatively uniform deep hydration state. On this basis, in step S52, 5% of the total amount of lycopene oil phase is taken and stirred at 800 r / min for 8 min to make a small amount of homologous oil phase evenly contact the particle surface and wet its natural hydrophobic region, while avoiding the formation of a large number of new oil droplets in the pre-activation stage. After standing in the dark for 15 min, the hydration layer and the oil phase pre-wetting layer on the particle surface reach a relative equilibrium. On this basis, in step S52, 5% of the total amount of lycopene oil phase is taken and stirred at 800 r / min for 8 min to make a small amount of oil phase fully contact the particle surface and improve the wettability of the particles to the oil phase. After standing in the dark for 15 min, the particles and the oil phase reach a relatively stable contact state. This treatment facilitates the migration and adsorption of particles to the interface of newly formed oil droplets during subsequent emulsification. However, the specific interfacial interaction mechanism is still related to the particle surface composition, wettability, and dispersion state. This allows for faster migration and adsorption onto the surface of newly formed oil droplets during subsequent batch emulsification, matching the particle adsorption rate with the oil droplet formation rate and providing a basis for forming a more continuous particle interface coverage structure.

[0016] Furthermore, in step S6, the volume ratio of the total amount of lycopene oil phase obtained in step S4 to the volume ratio of the whole biomass particle dispersion obtained in step S3 is 3:7, 4:6, 5:5, 6:4 or 7:3.

[0017] It should be noted that the above five oil-water volume ratios are adjusted stepwise by varying the lycopene oil phase loading and the ratio of the continuous water phase to match the interfacial area of ​​the newly formed oil droplets, the number of whole biomass particles that can participate in interfacial adsorption, and the viscoelasticity of the continuous phase. This results in an adjustable and stable range between lycopene loading capacity, particle interfacial coverage, and emulsion processing fluidity.

[0018] Furthermore, in step S6, 5% of the total amount of lycopene oil phase is used for the pre-wetting treatment in step S5, and the remaining lycopene oil phase is divided into a first oil phase, a second oil phase, and a third oil phase according to 30%, 35%, and 30% of the total amount of lycopene oil phase. After adding the first oil phase, shear at 10000 r / min for 60 s and let stand for 5 min in the dark. After adding the second oil phase, shear at 12000 r / min for 60 s and let stand for 5 min in the dark. After adding the third oil phase, the mixture was sheared at 12000 r / min for 90 s and then allowed to stand for 5 min in the dark. After adding all the lycopene oil phase, the final granulation process was carried out at 10000 r / min for 60 s; In step S7, the low-shear interface rearrangement treatment involves stirring the lycopene Pickering emulsion at 150 r / min for 15 min at 25°C under light-protected conditions.

[0019] It should be noted that the 5% lycopene oil phase used in step S5 pre-imparts suitable oil-water interface affinity to the whole biomass particles. The remaining 95% oil phase is added in three batches of 30%, 35%, and 30% to match the rate of increase in oil droplet interface area with the rate of particle migration, adsorption, and film formation. After the first 30% oil phase is added, it is sheared at 10,000 r / min for 60 s to allow the pre-activated particles to participate in the formation and coverage of the initial oil droplets. Then, it is allowed to stand for 5 min to allow the particles to complete interfacial adsorption and rearrangement of the primary particle layer. On the basis of the formation of the primary interface structure, the second 35% oil phase is added, and the shearing speed is increased to 12,000 r / min for 60 s to adapt to the increase in system viscosity and the area of ​​the interface to be stabilized after the increase of oil phase. Then, it is allowed to stand for 5 min to allow the newly formed oil droplets to be covered by particles in time. The third oil phase is controlled at 30% to limit the instantaneous increase in interface area in the later stage of emulsification, and the shearing is extended to 90% at 12,000 r / min. Step S7 involves stirring the emulsion at 150 r / min for 15 min under conditions of high oil content and viscosity to fully disperse the remaining oil phase. After standing for 5 min, the remaining oil phase is fully dispersed, and then allowed to stand for 5 min to complete the interfacial film formation of the last batch of oil droplets. After all the oil phase is added, the emulsion is granulated at 10,000 r / min for 60 s to homogenize the remaining coarse oil droplets and local aggregate structures, while reducing the disturbance of the formed particle interfacial layer caused by continuous high-intensity shear. On this basis, step S7 involves stirring the emulsion at 150 r / min for 15 min at 25℃ under light-protected conditions to perform low-shear interfacial rearrangement treatment. This keeps the formed oil droplets in a relatively uniform dispersion state and provides conditions for further rearrangement of interfacial particles and stability of the interfacial structure. This is conducive to the formation of a more continuous particle interfacial coverage structure, reduces the structural disturbance of the emulsion in subsequent processing, and reduces the risk of lycopene degradation caused by light exposure during the processing.

[0020] This application also provides a lycopene Pickering emulsion stabilized by whole biomass particles of Pleurotus ostreatus mycelium, an O / W type Pickering emulsion stabilized by whole biomass particles of edible fungi mycelium, comprising a continuous aqueous phase, lycopene-containing oil droplets dispersed in the continuous aqueous phase, and whole biomass particles of edible fungi mycelium located at the interface region between the oil droplets and the continuous aqueous phase, wherein the whole biomass particles of edible fungi mycelium are used to form a particle covering structure at the oil droplet interface; the lycopene Pickering emulsion is prepared by the above preparation method.

[0021] It should be noted that the lycopene Pickering emulsion is not a simple mixture of lycopene oil phase, continuous aqueous phase, and mycelial particles. Rather, it is an interface assembly system with a specific spatial distribution formed through the above-mentioned hydration dispersion, interface pre-activation, batch emulsification, and interface rearrangement processes. In this system, lycopene is dissolved or dispersed inside the oil droplets. The whole biomass particles of Pleurotus eryngii mycelium utilize their naturally coexisting proteins, polysaccharides, dietary fiber, chitin, and cell wall structures to form hydrophilic and hydrophobic regions, which are directionally adsorbed at the interface between the lycopene oil droplets and the continuous aqueous phase. After arrangement, intercalation, and locking, they form a particle interface layer that is difficult to detach, thereby creating spatial barriers to adjacent oil droplets and restricting their migration and aggregation. At the same time, it allows the lycopene oil phase to be uniformly dispersed in the continuous aqueous phase in the form of oil droplets, which helps to reduce emulsion stratification and oil separation. It may also provide a certain physical shielding effect on lycopene through the oil droplet and interface particle structure.

[0022] The core invention of this application lies in using whole biomass particles of edible fungi mycelium that have not undergone protein or polysaccharide extraction, enzymatic purification, or chemical modification as an interface stabilizer. It sequentially couples interface functionalization dispersion through low-speed pre-dispersion-high-speed shearing and cooling cycles, interface pre-activation through vacuum pulse rehydration-pre-wetting with a small amount of lycopene oil phase, segmented film-locking emulsification through batch oiling-short-time shearing-intermittent settling-final granulation, and low-shear interface rearrangement after emulsification. This allows porous, fibrous, and irregular mycelial particles to undergo sufficient hydration and physical dispersion during oil droplet formation. Through oil phase pre-wetting and batch emulsification, the particles migrate to the interface region of the newly formed oil droplets, where they undergo adsorption and rearrangement, overcoming the limitations of conventional methods. This method addresses the issues of particle adsorption lag, insufficient interface coverage, oil droplet aggregation, and widened particle size distribution that are easily caused by single-stage oiling and continuous high-speed shearing. It simplifies raw material processing and improves the overall utilization rate of mycelium while forming a more continuous particle interface coverage structure, enhancing the anti-aggregation, anti-stratification, and anti-oil separation capabilities of the lycopene Pickering emulsion. Performance tests further show that the particle mass-volume concentration is approximately 5%–6%, with 6% exhibiting superior lycopene encapsulation and storage retention performance. An oil-water ratio of 5:5–6:4 is most suitable, and the resulting O / W emulsion demonstrates good comprehensive performance in terms of droplet uniformity, emulsification stability, lycopene encapsulation, and storage retention. Attached Figure Description

[0023] Figure 1 Image showing the appearance stability of lycopene Pickering emulsions with different concentrations of whole biomass particles of Pleurotus ostreatus mycelium and oil-water ratios during storage at 0d, 7d and 14d. Figure 2 Optical microscope images of lycopene Pickering emulsions at different particle concentrations and oil-water ratios; Figure 3Confocal laser scanning microscopy images of lycopene Pickering emulsions with particle concentrations of 4%, 5%, and 6% at different oil-water ratios; Figure 4 Figure 1 shows the results of Emulsion Stability Index (ESI) and Emulsifying Activity Index (EAI) of lycopene Pickering emulsions under different particle concentrations and oil-water ratios. Figure 5 The three-phase contact angle test diagrams are shown for interfacial particles at different oil-water ratios with particle concentrations of 4%, 5%, and 6%. Figure 6 Figure 1 shows the test results of lycopene encapsulation efficiency and storage retention rate under different particle concentrations and oil-water ratios; Figure 7 The graph shows the results of shear thinning, frequency scanning, and thixotropic recovery tests of a lycopene Pickering emulsion with an oil-water ratio of 5:5 and a particle concentration of 3%–7%. Figure 8 The shear stress and apparent viscosity of the whole biomass particle aqueous dispersion reference group and the lycopene Pickering emulsions with different oil-water ratios are shown as changes with shear rate. Figure 9 The graph shows the particle size distribution, zeta potential, three-phase contact angle, and dynamic changes of oil-water interfacial tension of the whole biomass particles of Pleurotus eryngii mycelium. Detailed Implementation

[0024] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0025] To investigate the effects of the mass concentration of Pleurotus eryngii mycelium biomass particles in the initial biomass particle dispersion and the volume ratio of lycopene oil phase to particle aqueous phase on the performance of lycopene Pickering emulsions, the particle mass concentrations were set to 3 g / 100 mL, 4 g / 100 mL, 5 g / 100 mL, 6 g / 100 mL and 7 g / 100 mL, respectively. Under the above particle mass concentration conditions, the volume ratios of lycopene oil phase to particle aqueous phase were set to 3∶7, 4∶6, 5∶5, 6∶4 and 7∶3, respectively, resulting in the preparation of 25 groups of lycopene Pickering emulsions.

[0026] Wherein, the particle mass concentration refers to the mass of Pleurotus eryngii mycelium whole biomass particles contained in every 100 mL of initial whole biomass particle dispersion; the particle aqueous phase refers to the whole biomass particle dispersion obtained in step S3; the oil-water ratio refers to the volume ratio of the total amount of lycopene oil phase obtained in step S4 to the whole biomass particle dispersion obtained in step S3; each emulsion group is calculated as 100 mL based on the sum of the volumes of the lycopene oil phase and the particle aqueous phase. Except for the particle mass concentration and oil-water ratio, the types of raw materials, lycopene oil phase concentration, particle hydration conditions, shear-cooling conditions, vacuum pulse rehydration conditions, oil phase pre-wetting conditions, batch emulsification conditions, terminal granulation conditions, and low-shear interface rearrangement conditions used in each group are kept consistent.

[0027] Example 1: Lycopene Pickering emulsion with a particle mass concentration of 3 g / 100 mL This embodiment provides a set of lycopene Pickering emulsions with different oil-water ratios, containing 3 g / 100 mL of whole biomass particles from Pleurotus eryngii mycelium in an initial whole biomass particle dispersion, including the following steps: S1. Take the whole biomass of Pleurotus eryngii mycelium, wash it three times with deionized water to remove culture medium residue and visible impurities; centrifuge at 4000 r / min for 10 min, and collect the mycelial precipitate; place the obtained mycelial precipitate in a 40℃ forced-air drying oven to constant weight, then pulverize and sieve to collect Pleurotus eryngii mycelium whole biomass particles with a particle size of 75–150 μm. The obtained particles retain the natural components of Pleurotus eryngii mycelium, such as protein, polysaccharide, dietary fiber, chitin, and cell wall fragments, without protein extraction, polysaccharide extraction, enzymatic purification, or chemical modification.

[0028] S2. Weigh out the whole biomass particles of Pleurotus ostreatus mycelium according to Table 1, add them to deionized water and make up to the corresponding particle aqueous phase volume, stir and hydrate at 600 r / min for 60 min to allow the particles to absorb water and swell, and obtain an initial whole biomass particle dispersion with a particle mass concentration of 3 g / 100 mL.

[0029] Table 1. Raw material usage for emulsions with different oil-to-water ratios in Example 1

[0030] S3. Process each initial whole biomass particle dispersion according to the following steps: S31. The initial whole biomass particle dispersion is pre-dispersed at 600 r / min for 10 min to obtain a pre-dispersed solution; S32. The pre-dispersed liquid is subjected to high-speed shearing at 16000 r / min for 5 min to obtain a sheared dispersion; S33. Cool the shear dispersion in an ice bath for 3 min; Steps S32 and S33 constitute a shear-cooling cycle, which is performed twice to obtain a whole biomass particle dispersion.

[0031] S4. Weigh the raw materials according to the mass ratio of lycopene to sunflower seed oil of 0.10:99.90, stir at 500 r / min for 30 min under light-protected conditions to make the lycopene uniformly dissolved or dispersed in the sunflower seed oil, and obtain a lycopene oil phase with a lycopene mass percentage of 0.10%.

[0032] S5. Perform interface pre-activation treatment on each whole biomass particle dispersion according to the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08 MPa and maintain for 3 min, restore to normal pressure and let stand for 3 min. Repeat the above evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase in each group and add it to the corresponding whole biomass particle dispersion after vacuum pulse rehydration treatment. For Examples 1-1 to 1-5, add 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of lycopene oil phase respectively, stir at 800 r / min for 8 min, and then let stand for 15 min in the dark to obtain a pre-activated Pleurotus eryngii mycelium whole biomass particle dispersion.

[0033] Table 2. Pre-wetting amount and batch addition amount of lycopene oil phase under different oil-water ratio conditions in Example 1

[0034] In Table 2, the amounts of the pre-wetting oil phase, the first oil phase, the second oil phase, and the third oil phase are 5%, 30%, 35%, and 30% of the total amount of lycopene oil phase, respectively. The sum of the amounts of the four oil phases is the total amount of lycopene oil phase. The data in each row correspond to Examples 1-1 to Examples 1-5 in sequence.

[0035] S6. According to the dosages shown in Table 2, the remaining lycopene oil phase after pre-wetting treatment in each group was divided into three oil phases: the first oil phase, the second oil phase, and the third oil phase. After adding the first oil phase, it was sheared at 10,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the second oil phase, it was sheared at 12,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the third oil phase, it was sheared at 12,000 r / min for 90 s, followed by standing in the dark for 5 min. After adding all the lycopene oil phase, it was subjected to terminal granulation treatment at 10,000 r / min for 60 s to obtain the lycopene Pickering emulsion.

[0036] S7. Each group of lycopene Pickering emulsions was placed at 25°C and in the dark, and stirred at a low speed of 150 r / min for 15 min to obtain the lycopene Pickering emulsions of Pleurotus eryngii mycelium whole biomass particles stabilized in Examples 1-1 to 1-5.

[0037] Example 2: Lycopene Pickering emulsion with a particle mass concentration of 4 g / 100 mL This embodiment provides a set of lycopene Pickering emulsions with different oil-water ratios, containing 4 g / 100 mL of whole biomass particles of Pleurotus ostreatus mycelium in an initial whole biomass particle dispersion, including the following steps.

[0038] S1. Take the whole biomass of Pleurotus ostreatus mycelium and wash it three times with deionized water; centrifuge it at 4000 r / min for 10 min and collect the mycelium precipitate; place the obtained mycelium precipitate in a 40℃ forced-air drying oven and dry it to constant weight, then crush and sieve it to collect Pleurotus ostreatus mycelium whole biomass particles with a particle size of 75-150 μm.

[0039] S2. Weigh out the whole biomass particles of Pleurotus ostreatus mycelium according to Table 3, add them to deionized water and make up to the corresponding particle aqueous phase volume, stir and hydrate at 600 r / min for 60 min to obtain an initial whole biomass particle dispersion with a particle mass concentration of 4 g / 100 mL.

[0040] Table 3. Raw material usage for emulsions with different oil-to-water ratios in Example 2

[0041] S3. Each initial whole biomass particle dispersion was pre-dispersed at 600 r / min for 10 min, then sheared at 16000 r / min for 5 min, followed by ice bath cooling for 3 min; the high-speed shearing and ice bath cooling were combined as a shear-cooling cycle, and the cycle was performed twice to obtain the whole biomass particle dispersion.

[0042] S4. Weigh the raw materials according to the mass ratio of lycopene to sunflower seed oil of 0.10:99.90, stir at 500 r / min for 30 min under light-protected conditions to obtain a lycopene oil phase with a lycopene mass percentage of 0.10%.

[0043] S5. Perform interface pre-activation treatment on each whole biomass particle dispersion according to the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08 MPa and maintain for 3 min, restore to normal pressure and let stand for 3 min. Repeat the above evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase in each group and add it to the corresponding whole biomass particle dispersion after vacuum pulse rehydration treatment. For Examples 2-1 to 2-5, add 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of lycopene oil phase respectively, stir at 800 r / min for 8 min, and then let stand for 15 min in the dark to obtain a pre-activated Pleurotus eryngii mycelium whole biomass particle dispersion.

[0044] Table 4. Pre-wetting amount and batch addition amount of lycopene oil phase under different oil-water ratio conditions in Example 2

[0045] In Table 4, the amounts of the pre-wetting oil phase, the first oil phase, the second oil phase, and the third oil phase are 5%, 30%, 35%, and 30% of the total amount of lycopene oil phase, respectively. The sum of the amounts of the four oil phases is the total amount of lycopene oil phase. The data in each row correspond to Examples 2-1 to 2-5 in sequence.

[0046] S6. Following the dosages shown in Table 4, the remaining lycopene oil phase after pre-wetting treatment in each group was sequentially divided into three oil phases: the first oil phase, the second oil phase, and the third oil phase. After adding the first oil phase, it was sheared at 10,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the second oil phase, it was sheared at 12,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the third oil phase, it was sheared at 12,000 r / min for 90 s, followed by standing in the dark for 5 min. After adding all the lycopene oil phase, terminal granulation was performed at 10,000 r / min for 60 s to obtain the lycopene Pickering emulsion.

[0047] S7. Each emulsion was stirred at 150 r / min for 15 min at 25°C in the dark to obtain the lycopene Pickering emulsions of Pleurotus ostreatus mycelium whole biomass particles stabilized in Examples 2-1 to 2-5.

[0048] Example 3: Lycopene Pickering emulsion with a particle mass concentration of 5 g / 100 mL This embodiment provides a set of lycopene Pickering emulsions with different oil-water ratios, containing 5 g / 100 mL of whole biomass particles of Pleurotus ostreatus mycelium in an initial whole biomass particle dispersion, including the following steps.

[0049] S1. Take the whole biomass of Pleurotus ostreatus mycelium, wash it three times with deionized water, centrifuge at 4000 r / min for 10 min to dehydrate it, dry it at 40℃ with forced air until constant weight, crush it and sieve it to obtain Pleurotus ostreatus mycelium whole biomass particles with a particle size of 75-150 μm.

[0050] S2. Weigh out the whole biomass particles of Pleurotus ostreatus mycelium according to Table 5, add them to deionized water and make up to the corresponding particle aqueous phase volume, stir and hydrate at 600 r / min for 60 min to obtain an initial whole biomass particle dispersion with a particle mass concentration of 5 g / 100 mL.

[0051] Table 5. Raw material dosage for emulsions with different oil-to-water ratios in Example 3

[0052] S3. Each initial whole biomass particle dispersion was pre-dispersed at 600 r / min for 10 min, then sheared at 16000 r / min for 5 min, followed by cooling in an ice bath for 3 min; the shear-cooling cycle was performed twice to obtain the whole biomass particle dispersion.

[0053] S4. Weigh the raw materials according to the mass ratio of lycopene to sunflower seed oil of 0.10:99.90, stir at 500 r / min for 30 min under light-protected conditions to obtain a lycopene oil phase with a lycopene mass percentage of 0.10%.

[0054] S5. Perform interface pre-activation treatment on each whole biomass particle dispersion according to the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08 MPa and maintain for 3 min, restore to normal pressure and let stand for 3 min. Repeat the above evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase in each group and add it to the corresponding whole biomass particle dispersion after vacuum pulse rehydration treatment. For Examples 3-1 to 3-5, add 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of lycopene oil phase sequentially, stir at 800 r / min for 8 min, and then let stand for 15 min in the dark to obtain a pre-activated Pleurotus eryngii mycelium whole biomass particle dispersion.

[0055] Table 6. Pre-wetting amount and batch addition amount of lycopene oil phase under different oil-water ratio conditions in Example 3

[0056] In Table 6, the amounts of the pre-wetting oil phase, the first oil phase, the second oil phase, and the third oil phase are 5%, 30%, 35%, and 30% of the total amount of lycopene oil phase, respectively. The sum of the amounts of the four oil phases is the total amount of lycopene oil phase. The data in each row correspond to Examples 3-1 to 3-5 in sequence.

[0057] S6. Following the dosages shown in Table 6, the remaining lycopene oil phase after pre-wetting treatment in each group was sequentially divided into three oil phases: the first oil phase, the second oil phase, and the third oil phase. After adding the first oil phase, it was sheared at 10,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the second oil phase, it was sheared at 12,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the third oil phase, it was sheared at 12,000 r / min for 90 s, followed by standing in the dark for 5 min. After adding all the lycopene oil phase, it was subjected to terminal granulation at 10,000 r / min for 60 s to obtain the lycopene Pickering emulsion.

[0058] S7. Each emulsion was stirred at 150 r / min for 15 min at 25°C in the dark to obtain the lycopene Pickering emulsions of Pleurotus ostreatus mycelium whole biomass particles stabilized in Examples 3-1 to 3-5.

[0059] Example 4: Lycopene Pickering emulsion with a particle mass concentration of 6 g / 100 mL This embodiment provides a set of lycopene Pickering emulsions with different oil-water ratios, containing 6 g / 100 mL of whole biomass particles of Pleurotus eryngii mycelium in an initial whole biomass particle dispersion, including the following steps.

[0060] S1. Take the whole biomass of Pleurotus ostreatus mycelium, wash it three times with deionized water, centrifuge at 4000 r / min for 10 min to dehydrate it, dry it at 40℃ with forced air until constant weight, crush it and sieve it to obtain Pleurotus ostreatus mycelium whole biomass particles with a particle size of 75-150 μm.

[0061] S2. Weigh out the whole biomass particles of Pleurotus ostreatus mycelium according to Table 7, add them to deionized water and make up to the corresponding volume of the particle aqueous phase, stir and hydrate at 600 r / min for 60 min to obtain an initial whole biomass particle dispersion with a particle mass concentration of 6 g / 100 mL.

[0062] Table 7 Raw material dosage for emulsions with different oil-water ratios in Example 4

[0063] S3. Each initial whole biomass particle dispersion was pre-dispersed at 600 r / min for 10 min, then sheared at 16000 r / min for 5 min, followed by cooling in an ice bath for 3 min; the shear-cooling cycle was performed twice to obtain the whole biomass particle dispersion.

[0064] S4. Weigh the raw materials according to the mass ratio of lycopene to sunflower seed oil of 0.10:99.90, stir at 500 r / min for 30 min under light-protected conditions to obtain a lycopene oil phase with a lycopene mass percentage of 0.10%.

[0065] S5. Perform interface pre-activation treatment on each whole biomass particle dispersion according to the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08 MPa and maintain for 3 min, restore to normal pressure and let stand for 3 min. Repeat the above evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase in each group and add it to the corresponding whole biomass particle dispersion after vacuum pulse rehydration treatment. For Examples 4-1 to 4-5, add 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of lycopene oil phase respectively, stir at 800 r / min for 8 min, and then let stand for 15 min in the dark to obtain a pre-activated Pleurotus eryngii mycelium whole biomass particle dispersion.

[0066] Table 8. Pre-wetting amount and batch addition amount of lycopene oil phase under different oil-water ratio conditions in Example 4

[0067] In Table 8, the amounts of the pre-wetting oil phase, the first oil phase, the second oil phase, and the third oil phase are 5%, 30%, 35%, and 30% of the total amount of lycopene oil phase, respectively. The sum of the amounts of the four oil phases is the total amount of lycopene oil phase. The data in each row correspond to Examples 4-1 to 4-5 in sequence.

[0068] S6. Following the dosages shown in Table 8, the remaining lycopene oil phase after pre-wetting treatment in each group was sequentially divided into three oil phases: the first oil phase, the second oil phase, and the third oil phase. After adding the first oil phase, it was sheared at 10,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the second oil phase, it was sheared at 12,000 r / min for 60 s, followed by standing in the dark for 5 min. After adding the third oil phase, it was sheared at 12,000 r / min for 90 s, followed by standing in the dark for 5 min. After adding all the lycopene oil phase, terminal granulation was performed at 10,000 r / min for 60 s to obtain the lycopene Pickering emulsion.

[0069] S7. Each emulsion was stirred at 150 r / min for 15 min at 25°C in the dark to obtain the lycopene Pickering emulsions of Pleurotus ostreatus mycelium whole biomass particles stabilized in Examples 4-1 to 4-5.

[0070] Example 5: Lycopene Pickering emulsion with a particle mass concentration of 7 g / 100 mL This embodiment provides a set of lycopene Pickering emulsions with different oil-water ratios, containing 7 g / 100 mL of whole biomass particles of Pleurotus eryngii mycelium in an initial whole biomass particle dispersion, including the following steps.

[0071] S1. Take the whole biomass of Pleurotus ostreatus mycelium, wash it three times with deionized water, centrifuge at 4000 r / min for 10 min to dehydrate it, dry it at 40℃ with forced air until constant weight, crush it and sieve it to obtain Pleurotus ostreatus mycelium whole biomass particles with a particle size of 75-150 μm.

[0072] S2. Weigh out the whole biomass particles of Pleurotus ostreatus mycelium according to Table 9, add them to deionized water and make up to the corresponding particle aqueous phase volume, stir and hydrate at 600 r / min for 60 min to obtain an initial whole biomass particle dispersion with a particle mass concentration of 7 g / 100 mL.

[0073] Table 9. Raw material usage for emulsions with different oil-to-water ratios in Example 5

[0074] S3. Each initial whole biomass particle dispersion was pre-dispersed at 600 r / min for 10 min, then sheared at 16000 r / min for 5 min, followed by cooling in an ice bath for 3 min; the shear-cooling cycle was performed twice to obtain the whole biomass particle dispersion.

[0075] S4. Weigh the raw materials according to the mass ratio of lycopene to sunflower seed oil of 0.10:99.90, stir at 500 r / min for 30 min under light-protected conditions to obtain a lycopene oil phase with a lycopene mass percentage of 0.10%.

[0076] S5. Perform interface pre-activation treatment on each whole biomass particle dispersion according to the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08 MPa and maintain for 3 min, restore to normal pressure and let stand for 3 min. Repeat the above evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase in each group and add it to the corresponding whole biomass particle dispersion after vacuum pulse rehydration treatment. For Examples 5-1 to 5-5, add 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL, and 3.5 mL of lycopene oil phase sequentially, stir at 800 r / min for 8 min, and then let stand for 15 min in the dark to obtain a pre-activated Pleurotus eryngii mycelium whole biomass particle dispersion.

[0077] Table 10. Pre-wetting amount and batch addition amount of lycopene oil phase under different oil-water ratio conditions in Example 5

[0078] In Table 10, the amounts of the pre-wetting oil phase, the first oil phase, the second oil phase, and the third oil phase are 5%, 30%, 35%, and 30% of the total amount of lycopene oil phase, respectively. The sum of the amounts of the four oil phases is the total amount of lycopene oil phase. The data in each row correspond to Examples 5-1 to 5-5 in sequence.

[0079] S6. According to the dosages shown in Table 10, the remaining lycopene oil phase after pre-wetting treatment in each group was divided into three oil phases: the first oil phase, the second oil phase, and the third oil phase. After adding the first oil phase, it was sheared at 10,000 r / min for 60 s, and then allowed to stand in the dark for 5 min. After adding the second oil phase, it was sheared at 12,000 r / min for 60 s, and then allowed to stand in the dark for 5 min. After adding the third oil phase, it was sheared at 12,000 r / min for 90 s, and then allowed to stand in the dark for 5 min. After adding all the lycopene oil phase, it was subjected to terminal granulation treatment at 10,000 r / min for 60 s to obtain the lycopene Pickering emulsion.

[0080] S7. Each emulsion was stirred at 150 r / min for 15 min at 25°C in the dark to obtain the lycopene Pickering emulsions of Pleurotus ostreatus mycelium whole biomass particles stabilized in Examples 5-1 to 5-5.

[0081] A total of 25 lycopene Pickering emulsions were obtained in Examples 1 to 5 above. The resulting emulsions used an aqueous phase as the continuous phase and sunflower seed oil droplets containing lycopene as the dispersed phase. Pleurotus eryngii mycelium biomass particles were distributed at the interface between the lycopene oil droplets and the continuous aqueous phase. These 25 emulsions were used for subsequent performance testing.

[0082] Performance testing methods In the following tests, Examples 1-1 to 1-5, Examples 2-1 to 2-5, Examples 3-1 to 3-5, Examples 4-1 to 4-5, and Examples 5-1 to 5-5 correspond to emulsions with particle concentrations of 3%, 4%, 5%, 6%, and 7%, respectively; sub-numbers 1 to 5 of each example correspond to oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3, respectively. Unless otherwise stated, all test samples are prepared according to the above relationships and corresponding appendices. Figure 1 One-to-one correspondence.

[0083] 1. Emulsion Appearance Stability Test—Corresponding Figure 1 Take 20 mL of each emulsion and place it in a transparent sealed sample bottle of the same specification. Store it at 25±1℃ in the dark. Take photos of the appearance of the emulsion on day 0, day 7 and day 14 of storage. Figure 1 The samples were divided into five groups based on particle concentration: 3%, 4%, 5%, 6%, and 7%. The five samples in each particle concentration group corresponded to oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3 from left to right. The three rows of photos corresponded to the appearance of the emulsion after storage for 0 days, 7 days, and 14 days.

[0084] Combined with Examples 1 to 5 and Figure 1 It can be seen that when stored for 0 days, the emulsions prepared under the conditions of particle mass volume concentration and oil-water ratio are uniformly orange-red and have not yet shown obvious macroscopic stratification. After 7 days of storage, some samples gradually showed an upper emulsion layer and a lower precipitate layer. As the storage time was extended to 14 days, the precipitate or stratification phenomenon became more obvious, indicating that the macroscopic appearance stability of the emulsion decreased with the extension of storage time. In comparison, samples with a particle volume concentration of 5%–6% and an oil-water ratio of 5:5 or 6:4 exhibited relatively better emulsion continuity and uniformity, with less localized flocculation. This suggests that appropriate particle addition is beneficial for improving the macroscopic stability of the emulsion, and its effect may be related to particle coverage at the oil droplet interface and changes in the continuous phase structure. When the particle volume concentration was 3%–4%, the particle supply was relatively insufficient, and some samples showed obvious separation or stratification earlier. When the particle volume concentration increased to 7%, more obvious flocculent or lumpy aggregation appeared in the upper emulsion layer after 7 and 14 days of storage, possibly due to enhanced interparticle interactions and increased system viscosity. Therefore, within the range investigated in this experiment, both excessively low and excessively high particle concentrations are detrimental to maintaining good appearance stability of the emulsion during a 14-day storage period. When the particle volume concentration is 5%–6% and the oil-water ratio is 5:5–6:4, there is a more suitable match between the particle supply and the requirements for oil-water interface stability.

[0085] 2. Optical microscope observation—corresponding Figure 2 Take 10 μL of each emulsion and drop it onto a glass slide. After covering with a coverslip, observe the morphology, size and distribution of the emulsion droplets under an optical microscope. Take pictures of no less than 5 fields of view for each group. The scale bar in the figure is 100 μm. Figure 2 The images were grouped according to particle concentrations of 3%, 4%, 5%, 6%, and 7%, with the five images in each group corresponding to oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3, respectively.

[0086] Combined with Examples 1 to 5 and Figure 2It is evident that the mass-volume concentration of the whole biomass particles of Pleurotus eryngii mycelium and the volume ratio of lycopene oil phase to particle aqueous phase jointly affect the size, number, and distribution of emulsion droplets. In the 3%–4% low particle concentration samples corresponding to Examples 1 and 2, some fields of view showed fewer droplets, larger droplet sizes, and uneven particle size distribution, indicating that when the particle supply is relatively insufficient, it is difficult to cover the newly formed oil-water interface during shearing in a timely and sufficient manner, and droplets are prone to contact and aggregation. Samples with oil-water ratios of 3:7 or 7:3 in each particle concentration group also showed varying degrees of sparse droplets, large size differences, or local aggregation, indicating that when the oil-water ratio deviates from the moderate range, the oil droplet formation process is related to the particle size distribution. The coordination between particle interfacial migration, adsorption, and the support of the continuous aqueous phase structure decreased. In contrast, in Examples 3-3, 3-4, 4-3, and 4-4, i.e., samples with a particle mass-volume concentration of 5%–6% and an oil-water ratio of 5:5 or 6:4, the oil droplet outlines were clearer, and the droplets were relatively small, dense, and uniformly distributed. This indicates that under these conditions, the particle supply and the oil droplet interfacial area had a good matching relationship, which could improve the interfacial coverage and inhibit droplet aggregation. In the 7% particle concentration sample corresponding to Example 5, more irregular dark areas and local aggregate structures were visible, indicating that excessively high particle concentration would increase the viscosity of the system and the interaction between particles, which was not conducive to uniform droplet dispersion. Therefore, the optical microscopy observation results further show that a particle mass-volume concentration of 5%–6% combined with an oil-water ratio of 5:5–6:4 is more conducive to the formation of lycopene Pickering emulsions with relatively small and uniformly distributed droplets.

[0087] 3. CLSM Observation—Correspondence Figure 3 This test selected 15 groups of emulsions with particle concentrations of 4%, 5%, and 6% for observation. Among them, Examples 2-1 to 2-5 correspond to a particle concentration of 4%, Examples 3-1 to 3-5 correspond to a particle concentration of 5%, and Examples 4-1 to 4-5 correspond to a particle concentration of 6%. Subgroups 1 to 5 in each group correspond to oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3, respectively. Figure 3 The three particle concentration groups correspond to 4%, 5%, and 6% respectively. Each group, from left to right, represents the five oil-water ratios mentioned above. Each sample, from top to bottom, represents the Nile Red channel, Nile Blue channel, and overlay image.

[0088] Take 1 mL of each emulsion, stain the lycopene oil phase with Nile Red, and label the whole biomass particles of Pleurotus eryngii mycelium and its related structures with Nile Blue. After incubating in the dark for 10 min, observe with a confocal laser scanning microscope and collect images of Nile Red channel, Nile Blue channel and the superimposed images of the two.

[0089] Combined with Examples 2 to 4 and Figure 3 It is evident that the Nile Red-labeled oil droplets and the Nile Blue-labeled mycelial particles are adjacent to each other. In the superimposed image, whole biomass particles can be observed distributed around the oil droplets, suggesting that the whole biomass particles of Pleurotus eryngii mycelium and their source structures can accumulate near the oil droplet interface and may participate in the stability of the oil-water interface. Among them, the samples with 5% to 6% particle concentration and 5:5 to 6:4 oil-water ratio corresponding to Examples 3-3, 3-4, 4-3 and 4-4 have relatively complete oil droplet outlines and relatively continuous interface particle signals, while the samples with oil-water ratios of 3:7 or 7:3 show uneven signal distribution or local interface discontinuity. This indicates that under medium particle concentration and oil-water ratio conditions, the distribution of particle-related fluorescence signals around the oil droplets is relatively continuous, suggesting that the interface particle coverage is relatively good under these conditions.

[0090] 4. Emulsifying activity index and emulsion stability index test—corresponding Figure 4 Immediately after the emulsions were prepared, samples were taken and diluted with a 0.1% sodium dodecyl sulfate solution. The initial absorbance A0 of the diluted solution at 500 nm was measured, and the emulsifying activity index EAI was calculated based on A0, dilution factor, particle concentration, and oil phase volume fraction. After the diluted solution was allowed to stand for 10 min, the absorbance A at 500 nm was measured again. 10 The Emulsion Stability Index (ESI) is calculated using the following formula: ESI(min)=A0×10÷(A0−A 10 ).

[0091] Three parallel samples were set up for each group. Figure 4 The top image shows the ESI test results, and the bottom image shows the EAI test results.

[0092] Combined with Examples 1 to 5 and Figure 4It is evident that, under the same oil-to-water ratio, EAI and ESI generally show a trend of first increasing and then decreasing with increasing particle mass volume concentration. Under the same particle mass volume concentration, the EAI and ESI of samples with a medium oil-to-water ratio are generally higher than those with an oil-to-water ratio of 3:7 or 7:3, indicating that particle concentration and oil-to-water ratio have a combined effect on emulsification performance. Among them, the samples in Examples 3-3, 3-4, 4-3, and 4-4, i.e., samples with a particle mass volume concentration of 5% to 6% and an oil-to-water ratio of 5:5 or 6:4, have both indicators at a relatively high level, with EAI and ESI of Example 3-3 being particularly outstanding. The 3% and 7% particle concentration groups corresponding to Examples 1 and 5, as well as some samples in each concentration group with an oil-to-water ratio of 3:7 or 7:3, have relatively low indicators. The results indicate that insufficient particle supply makes it difficult to fully cover the interface of newly formed oil droplets, while excessively high particle concentration enhances the interaction between particles and may lead to local aggregation. When the oil-water ratio deviates from the moderate range, the matching relationship between particle supply and oil droplet interface area will be affected, all of which are detrimental to improving emulsification activity and emulsion stability.

[0093] 5. Three-phase contact angle test—corresponding Figure 5 This test selected 15 groups of samples from Examples 2-1 to 2-5, Examples 3-1 to 3-5, and Examples 4-1 to 4-5, corresponding to particle mass concentrations of 4%, 5%, and 6%, respectively. Sub-numbers 1 to 5 in each of the above particle mass concentration groups correspond to oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3, respectively. Figure 5 From top to bottom, the images show particle mass concentrations of 4%, 5%, and 6%, with each row from left to right displaying the contact angle droplet images for the five oil-water ratios mentioned above.

[0094] The interfacial particles in each emulsion were separated, washed, and dried, and then pressed into granular sheets with a smooth surface. The granular sheets were placed in a sunflower seed oil environment, and 5 μL of deionized water was added to the surface of the granular sheets. After stabilizing for 30 s, the droplet profile was collected, and 5 different positions were measured for each group.

[0095] Combined with Examples 2 to 4 and Figure 5As can be seen, the apparent three-phase contact angles of the particles separated from the 15 groups of samples were all less than 90°, indicating that the particles as a whole exhibited a preferential wetting characteristic of the aqueous phase, which is consistent with the wetting tendency of forming O / W type Pickering emulsions. It should be noted that whether the particles can form a stable Pickering emulsion is also related to factors such as particle size, morphology, interfacial adsorption strength, and interfacial coverage. Therefore, the contact angle results are mainly used to characterize the wetting tendency of the particles. Among them, in Examples 3-3, 3-4, 4-3, and 4-4, that is, the samples with a particle mass-volume concentration of 5% to 6% and an oil-water ratio of 5:5 or 6:4, the water droplets on the particle surface showed a relatively suitable limited spreading state, indicating that the particles were neither completely spread by the aqueous phase nor excessively repelled by the aqueous phase. The degree of wetting of the particles by the oil and aqueous phases was relatively coordinated. This wetting state provides a basis for the adsorption of particles at the oil-water interface and the maintenance of the aqueous phase as a continuous phase, which is conducive to the formation of a stable O / W type particle interfacial coverage structure.

[0096] 6. Lycopene encapsulation and retention rate test—corresponding Figure 6 The total lycopene content in the emulsion and the non-oil droplet-related lycopene content in the continuous phase were determined using a standardized method of demulsification, centrifugation, and organic solvent extraction. All samples underwent the same number of extractions, extractant volumes, and volume adjustment methods, and the lycopene extraction recovery rate was verified through a standard recovery test. The total lycopene content (M) in each group of emulsions was determined. 总 And the lycopene content M in the continuous phase that is not encapsulated by oil droplets 游离 The lycopene encapsulation rate is calculated using the following formula: Encapsulation efficiency (%) = (M 总 -M 游离 )÷M 总 ×100%.

[0097] Each emulsion group was stored at 25±1℃ under light-protected conditions. Lycopene content was measured on storage days 0, 7, and 14, and the lycopene retention rate on day t was calculated using the following formula: Retention rate R t (%) = M t ÷M0×100%; In the formula, M0 is the lycopene content in the emulsion on storage day 0, M t The lycopene content in the emulsion on day t of storage is given, where t is 7 days or 14 days. Figure 6 The top image shows the lycopene encapsulation rate, and the bottom image shows the lycopene storage retention rate.

[0098] Combined with Examples 1 to 5 and Figure 6It is evident that Examples 4-3 and 4-4, i.e., samples with a particle mass-volume concentration of 6% and oil-water ratios of 5:5 and 6:4 respectively, exhibited high lycopene encapsulation rates. After storage for 7 and 14 days, the 6% particle concentration group corresponding to Example 4 showed relatively high lycopene retention levels under various oil-water ratio conditions, while the corresponding data for the 3% low particle concentration group corresponding to Example 1 were generally lower. Combined with... Figure 6 The results showed that different particle concentrations and oil-water ratios of *Pleurotus eryngii* mycelium biomass had a certain impact on the lycopene encapsulation efficiency and retention rate during storage. Specifically, Examples 4-3 and 4-4, with a particle concentration of 6% and oil-water ratios of 5:5 and 6:4, exhibited higher lycopene encapsulation efficiency. After 7 and 14 days of storage, the 6% particle concentration group showed relatively higher lycopene retention levels under various oil-water ratio conditions, while the 3% particle concentration group generally showed lower retention levels. These results indicate that, within the scope of this experiment, appropriately increasing the particle concentration is beneficial for improving the encapsulation of lycopene in the emulsion and its storage stability. This phenomenon may be related to the fact that a higher particle supply is conducive to forming more sufficient particle coverage at the oil droplet interface and enhancing the overall structural stability of the emulsion. However, when the particle concentration was further increased to 7%, the encapsulation efficiency and retention rate did not show a further synchronous increasing trend, indicating that there may be an optimal range between the particle addition amount and the structural stability of the emulsion.

[0099] 7. Rheological property testing of emulsions with an oil-to-water ratio of 5:5—corresponding to Figure 7 This test selected five groups of emulsions: Examples 1-3, 2-3, 3-3, 4-3, and 5-3. The oil-to-water ratio of the emulsions was 5:5, and the particle concentrations were 3%, 4%, 5%, 6%, and 7%, respectively. Figure 7 The shear thinning curves, G′ / G″ frequency scan curves, and thixotropic recovery curves were all compared according to the above five particle concentrations.

[0100] The tests were conducted using a rotational rheometer and a parallel plate fixture. The parallel plate diameter was 40 mm, the test gap was 1 mm, and the test temperature was 25℃. The shear rate range for the shear thinning test was 0.1–100 s⁻¹. -1 Before frequency scanning, an oscillatory strain scan was performed to determine the linear viscoelastic region of the system, and strain values ​​within the linear viscoelastic region were selected for frequency scanning; the frequency scanning range was 0.1–10 Hz. The thixotropic recovery test adopted a three-stage mode of low strain—high strain—low strain, with the strain value and duration remaining consistent in each stage, and the change of storage modulus G′ over time was recorded in each stage.

[0101] Combining Examples 1-3, 2-3, 3-3, 4-3, and 5-3, and Figure 7 It is evident that, under the condition of an oil-to-water ratio of 5:5, as the particle mass-volume concentration increased from 3% to 7%, the low-shear viscosity of the emulsion, as well as G′ and G″, generally increased. This indicates that with the increase of particle concentration, the overall structural strength and viscoelastic response of the system are enhanced, which may be related to the continuous phase structure formed by the particles and the enhanced interaction between oil droplets and particles. The apparent viscosity of each group of emulsions decreased with increasing shear rate, indicating that the emulsions exhibit significant shear-thinning behavior. Within the tested frequency range, G′ was generally higher than G″, indicating that the emulsions also possess a viscoelastic structure dominated by elastic response. In the three-stage thixotropic recovery test, G′ decreased significantly in the high-strain stage, indicating that the internal structure of the emulsion was damaged. After recovering to the low-strain stage, G′ increased again, indicating that the system has a certain structural recovery capability. Among them, the 5%–6% particle concentration samples corresponding to Examples 3-3 and 4-3 showed a good balance between structural strength, recovery capability, and processing fluidity. Although Example 5-3 had high viscosity and structural strength, the excessively high viscosity may affect the uniform mixing, transportation, and subsequent processing of the emulsion.

[0102] 8. Test on the relationship between shear rate, shear stress, and apparent viscosity—corresponding Figure 8 Figure 8 The data shown include a reference group of aqueous dispersions of whole biomass particles and five groups of emulsions with oil-water ratios of 3:7, 4:6, 5:5, 6:4 and 7:3, used to compare the changes in shear stress and apparent viscosity with shear rate between emulsions with different oil-water ratios and aqueous dispersions of whole biomass particles. Figure 8 Since the particles were not grouped according to a concentration of 3% to 7%, this test was only used to compare the rheological differences between the aqueous particle dispersion and emulsions with different oil-water ratios, and was not used to make intergroup comparisons of different particle concentrations.

[0103] Using a rotational rheometer, the shear rate was reduced from 0.1 s⁻¹ at 25°C. -1 Continuously increase to 100 s -1 Simultaneously, shear stress and apparent viscosity were recorded.

[0104] Combined with Examples 3-1 to 3-5 and Figure 8It is evident that, under the condition that the particle mass-volume concentration is 5%, compared with the reference group of the whole biomass particle aqueous dispersion obtained in step S3 of Example 3, the shear stress of the five emulsions with oil-water ratios of 3:7, 4:6, 5:5, 6:4, and 7:3 corresponding to Examples 3-1 to 3-5 all increased with increasing shear rate, while the apparent viscosity decreased with increasing shear rate, indicating that each group of emulsions exhibits pseudoplastic shear thinning characteristics. Among them, the 3:7 oil-water ratio emulsion corresponding to Example 3-1 has relatively low low shear viscosity and shear stress, indicating that when the oil phase ratio is low, the oil droplets aggregate. The structural relationships are relatively weak; the 7:3 oil-water ratio emulsions corresponding to Examples 3-5 have relatively high low-shear viscosity and shear stress, indicating that the higher oil phase volume fraction enhances the interaction between oil droplets, but may also increase the processing resistance of the system; the 5:5 and 6:4 oil-water ratio emulsions corresponding to Examples 3-3 and 3-4 have shear stress and apparent viscosity curves in the middle range and change relatively smoothly with shear rate, indicating that when the particle concentration is kept consistent, the medium oil-water ratio of 5:5 to 6:4 is more conducive to maintaining a better balance between the low-shear structural stability of the emulsion and the high-shear processing fluidity.

[0105] 9. Characterization of whole biomass particles—corresponding Figure 9 This test used the whole biomass particle dispersions obtained in step S3 of Examples 1 to 5 as the treatment group, corresponding to particle concentrations of 3%, 4%, 5%, 6% and 7%, respectively; and used the initial whole biomass particle dispersions that had not undergone the interface functionalization dispersion treatment in step S3 as the blank group. Figure 9 The four data points are, in order, particle size distribution, zeta potential, three-phase contact angle, and dynamic changes in oil-water interfacial tension from 0 to 600 s.

[0106] Before laser particle size and zeta potential testing, all samples were dispersed in the same medium and diluted to the instrument's recommended measurement range to ensure consistent test concentration and ionic strength. Each group underwent at least three independent measurements, and the average value was taken. After separating, drying, and pressing the particles into tablets, the three-phase contact angle of the particles was determined using the tablet pressing method. The dynamic interfacial tension between sunflower seed oil and the aqueous dispersion of the particles was determined using the pendant drop method within 0–600 s.

[0107] Combined with the whole biomass particle dispersion obtained in step S3 of Examples 1 to 5 and Figure 9As can be seen, Examples 1 to 5 correspond to treatment groups with particle mass-volume concentrations of 3%, 4%, 5%, 6%, and 7%, respectively. Compared with the blank group without step S3, the main peak particle size of the 3% to 5% treatment groups generally shifted towards smaller particle sizes, suggesting that high-speed shearing can promote the dissociation of hyphae and particle aggregates to a certain extent. The main peak particle size of the 6% and 7% treatment groups rebounded, indicating that factors such as particle re-aggregation or multiple scattering may affect the particle size test results at higher particle concentrations. Therefore, the effect of step S3 on particle size does not change monotonically with particle concentration. This indicates that after the low-speed pre-dispersion in step S31, the high-speed shearing in step S32, and the ice bath cooling in step S33 are implemented in the prescribed order and form a shear-cooling cycle, they can promote the dissociation of mycelial bundles and particle aggregates as a whole, making the resulting particle size more uniform. The above changes are the comprehensive result of the connection and cyclical effect of each operation. The Zeta potential of the particle dispersion in each treatment group is negative, and its absolute value increases with the increase of particle mass volume concentration, indicating that the particle surface has a certain negative charge, and suggesting that the particle dispersion state and interfacial charge may change with the treatment conditions. Since the Zeta potential is affected by particle concentration, ionic strength, pH, and measurement conditions, the Zeta potential result should not be used alone to judge the particle re-aggregation trend. It should be judged in conjunction with particle size distribution and other dispersion stability indicators. The three-phase contact angle of the particles increases with increasing particle concentration, but is less than 90°, indicating that the particles still maintain the preferential wetting characteristic of the aqueous phase and gradually form a more suitable oil-water two-phase wetting equilibrium, which is suitable for stabilizing O / W type Pickering emulsions. After adding the particles obtained in step S3, the oil-water interfacial tension decreased significantly within 0–600 s and gradually tended to stabilize. The dynamic interfacial tension results showed that, compared with the blank group, the oil-water interfacial tension decreased significantly and gradually tended to stabilize after adding the particles obtained in step S3 during the test, indicating that the mycelial whole biomass particles can affect the dynamic properties of the oil-water interface. The interfacial tension changes more significantly at higher particle concentrations, suggesting that the particle supply may affect the interface formation process. The results of particle size, Zeta potential, three-phase contact angle and dynamic interfacial tension corroborate each other, supporting that the whole biomass particles obtained in step S3 have good aqueous dispersibility, are suitable for forming O / W type emulsions, and have the ability to migrate to the oil-water interface and undergo adsorption.

[0108] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a lycopene Pickering emulsion with stabilized edible fungal mycelial particles, characterized in that, Includes the following steps: S1. Take the whole biomass of edible fungi mycelium, and after washing, dehydration, drying, crushing and sieving, obtain granules of whole biomass of edible fungi mycelium; S2. Add the edible fungus mycelium biomass particles to water and stir to hydrate them, to obtain an initial whole biomass particle dispersion; S3. The initial whole biomass particle dispersion is subjected to low-speed pre-dispersion, high-speed shear dispersion and ice bath cooling treatment in sequence to obtain the whole biomass particle dispersion. S4. Add lycopene to edible oil and stir to dissolve or disperse it under light-protected conditions to obtain the lycopene oil phase; S5. The whole biomass particle dispersion obtained in step S3 is subjected to vacuum pulse rehydration treatment. Part of the lycopene oil phase obtained in step S4 is added to the whole biomass particle dispersion that has been vacuum pulse rehydration treatment for pre-wetting treatment, so as to obtain the edible fungus mycelium whole biomass particle dispersion with pre-activated interface. S6. Add the remaining lycopene oil phase obtained in step S4 to the pre-activated edible fungus mycelium whole biomass particle dispersion obtained in step S5 in batches. After each batch of lycopene oil phase is added, short-time shearing and intermittent standing are performed in sequence. After all the lycopene oil phase is added, terminal granulation is performed to obtain lycopene Pickering emulsion. S7. The lycopene Pickering emulsion is subjected to low-shear interface rearrangement treatment to obtain a lycopene Pickering emulsion with stable whole biomass particles of edible fungi mycelium.

2. The preparation method according to claim 1, characterized in that, In step S1, the edible fungus mycelium is king oyster mushroom mycelium.

3. The preparation method according to claim 1, characterized in that, In step S2, the mass concentration of edible fungi mycelium whole biomass particles in the initial whole biomass particle dispersion is 3-7 g / 100 mL, and the stirring hydration time is 60 min.

4. The preparation method according to claim 1, characterized in that, Step S3 includes the following steps: S31. The initial whole biomass particle dispersion is pre-dispersed at 600 r / min for 10 min to obtain a pre-dispersed solution; S32. The pre-dispersed liquid is subjected to high-speed shearing at 16000 r / min for 5 min to obtain a sheared dispersion; S33. Cool the shear dispersion in an ice bath for 3 minutes; Steps S32 and S33 constitute a shear-cooling cycle, which is performed twice to obtain the whole biomass particle dispersion.

5. The preparation method according to claim 1, characterized in that, In step S4, the edible oil is sunflower seed oil, and the lycopene content in the lycopene oil phase is 0.10% by mass; the lycopene oil phase is prepared by stirring at 500 r / min for 30 min under light-protected conditions.

6. The preparation method according to claim 1, characterized in that, Step S5 includes the following steps: S51. Place the whole biomass particle dispersion obtained in step S3 into a sealed container, evacuate to −0.08MPa and maintain for 3min, restore to normal pressure and let stand for 3min. Repeat the evacuation, restoration to normal pressure and standing process 3 times to obtain the whole biomass particle dispersion after vacuum pulse rehydration treatment. S52. Take 5% of the total amount of lycopene oil phase obtained in step S4 and add it to the whole biomass particle dispersion after vacuum pulse rehydration treatment. Stir at 800 r / min for 8 min and then let stand for 15 min in the dark to obtain the pre-activated edible fungus mycelium whole biomass particle dispersion.

7. The preparation method according to claim 1, characterized in that, In step S6, the total volume ratio of the lycopene oil phase obtained in step S4 to the total volume ratio of the whole biomass particle dispersion obtained in step S3 is 3:7, 4:6, 5:5, 6:4 or 7:

3.

8. The preparation method according to claim 7, characterized in that, In step S6, 5% of the total amount of lycopene oil phase is used for the pre-wetting treatment in step S5, and the remaining lycopene oil phase is divided into a first oil phase, a second oil phase, and a third oil phase according to 30%, 35%, and 30% of the total amount of lycopene oil phase. After adding the first oil phase, shear at 10000 r / min for 60 s and let stand for 5 min in the dark. After adding the second oil phase, shear at 12000 r / min for 60 s and let stand for 5 min in the dark. After adding the third oil phase, the mixture was sheared at 12000 r / min for 90 s and then allowed to stand for 5 min in the dark. After adding all the lycopene oil phase, the final granulation process was carried out at 10000 r / min for 60 s; In step S7, the low-shear interface rearrangement treatment involves stirring the lycopene Pickering emulsion at 150 r / min for 15 min at 25°C under light-protected conditions.

9. A lycopene O / W type Pickering emulsion stabilized with whole biomass particles of edible fungi mycelium, characterized in that, The mixture comprises a continuous aqueous phase, lycopene-containing oil droplets dispersed in the continuous aqueous phase, and whole biomass particles of edible fungi mycelium located at the interface region between the oil droplets and the continuous aqueous phase, wherein the whole biomass particles of edible fungi mycelium are used to form a particle covering structure at the oil droplet interface; the lycopene Pickering emulsion is prepared by the preparation method according to any one of claims 1 to 8.

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