A fermenting beverage of a genus of the genus of the genus of the genus of the genus of the genus of the genus of the genus of the

Abstract

The application provides a fermenting beverage of Sparassis and a preparation method thereof, which is prepared by fermenting Sparassis extraction liquid with microcapsules. The microcapsules have an asymmetric structure, one side of which is a dense hydrophobic layer composed of Brazil palm wax, shea kernel oil and microcrystalline cellulose, and the inside of which is embedded with glucose, sodium bicarbonate and glucose oxidase, and the other side of which is a loose hydrophilic layer composed of inulin and a composite gel of gellan gum, and the inside of which is loaded with composite lactobacillus. The microcapsules can protect lactobacillus from the stress of inhibiting components and high viscosity polysaccharides in the extraction liquid in the early stage of fermentation, guarantee the rapid proliferation of the bacterial body, trigger the gas production reaction through the metabolic products, and realize self-driven fermentation. The application effectively breaks the mass transfer barrier, relieves the environmental stress, thereby maintaining the high activity of the bacterial body, and comprehensively improves the fermentation efficiency.

Description

Technical Field

[0001] This invention belongs to the field of food fermentation technology, specifically relating to a fermented beverage made from *Hydrangea macrophylla* and its preparation method. Background Technology

[0002] As a rare edible and medicinal fungus, *Hydrangea macrophylla* is rich in active ingredients such as β-glucan and has extremely high development value. Fermenting *Hydrangea macrophylla* with compound microbial agents to prepare functional beverages is an important direction for increasing its added value. However, this process faces three core technological bottlenecks:

[0003] 1. Inhibited cell activity: Certain natural antibacterial components or polyphenols and osmotic pressure contained in the *Hydrangea macrophylla* extract inhibit cell growth and metabolism, resulting in slow fermentation initiation and insufficient cell proliferation. Environmental stress caused by the accumulation of metabolic acid during the later stages of fermentation severely inhibits cell activity, leading to low fermentation efficiency.

[0004] 2. Low mass transfer efficiency: Polysaccharides in *Hydrangea macrophylla* (such as β-glucan) increase the viscosity of the extract, forming a dense liquid network. This severely hinders the diffusion of oxygen and nutrients, leading to a rapid deterioration of the microenvironment around the cells (nutrient depletion, accumulation of metabolic waste), and a significant reduction in fermentation efficiency.

[0005] 3. Uncontrollable and unstable fermentation process: Due to the above-mentioned inhibition and mass transfer problems, there are large differences between batches of traditional fermentation, making it difficult to accurately control the fermentation rate and endpoint acidity, which affects the uniformity and stability of product quality.

[0006] 4. Insufficient conversion of functional components: Insufficient contact between the bacteria and the macromolecular active substances of Hydrangea macrophylla (such as polysaccharides) limits the biotransformation of the bacteria, which may prevent the full release or generation of new functional factors, thus reducing the core value of the product.

[0007] 5. Existing encapsulation technologies (such as sodium alginate and chitosan microspheres) mostly provide only static physical isolation. Although they protect the cells to a certain extent, they often come at the cost of mass transfer efficiency, hindering the efficient transport of substrates, nutrients and metabolites, severely inhibiting cell activity and fermentation efficiency, and failing to intelligently regulate the fermentation process. Summary of the Invention

[0008] Technical Problem to be Solved: To address the aforementioned technical problems, the purpose of this invention is to provide a *Hydrangea macrophylla* fermented beverage and its preparation method. The beverage is prepared by fermenting *Hydrangea macrophylla* extract using microcapsules. The microcapsules have an asymmetric structure, with a dense hydrophobic layer on one side and a loose hydrophilic layer on the other. In the early stages of fermentation, the microcapsules effectively prevent contact between the bacteria and inhibitors in the external *Hydrangea macrophylla* extract. As lactic acid accumulates and bacteria proliferate during fermentation, a gas-producing reaction is triggered by enzymatic reactions within the core, increasing mass transfer efficiency and achieving self-driven fermentation.

[0009] Technical solution: A fermented beverage made from *Hydrangea macrophylla* extract using microcapsules. The microcapsules have an asymmetric structure, with one side being a dense hydrophobic layer composed of carnauba wax, shea butter, and microcrystalline cellulose, containing glucose, sodium bicarbonate, and glucose oxidase. The other side is a loose hydrophilic layer composed of inulin and gellan gum composite gel, containing composite lactobacillus.

[0010] The preparation method of the above-mentioned *Hydrangea macrophylla* fermented beverage includes the following steps:

[0011] S1. Select fresh, unrotten *Hydrangea macrophylla*, wash and tear into 1-3 cm pieces, add to water at 90-95℃ at a solid-liquid ratio of 1:4-10, extract for 60 min, filter through a 300-400 mesh filter cloth to obtain *Hydrangea macrophylla* extract.

[0012] S2. Add 7-15% of the mass of the hydrangea extract with white sugar, pasteurize in a constant temperature magnetic stirring water bath at 80-85℃ for 10-15 minutes, transfer to a sterile fermenter, and cool to room temperature for later use.

[0013] S3. Under aseptic conditions, add 0.8-2.5% by weight of the *Hydrangea macrophylla* extract microcapsules to the fermenter and let it ferment at 35-38°C for 36-50 hours.

[0014] S4. After fermentation, pasteurize in a constant temperature magnetic stirring water bath at 80~85℃ for 10~15 minutes, then fill and seal.

[0015] Preferably, the number of live bacteria in the microcapsules in step S3 is 7 × 10⁻⁶. 7 ~1×10 8 CFU / g.

[0016] The preparation method of microcapsules in step S3 above includes the following steps:

[0017] S31. Weigh out food-grade glucose, food-grade sodium bicarbonate and glucose oxidase, mix them evenly, and seal the mixed powder in a light-proof and moisture-proof container for later use.

[0018] S32. Weigh out carnauba wax and shea butter, heat them in a water bath to melt them, add sodium stearoyl lactylate, stir to form a homogeneous oil phase, then add microcrystalline cellulose and continue stirring to form a uniform hydrophobic wall material dispersion.

[0019] S33. Using the mixed powder prepared in S31 as the core, and the hydrophobic wall material dispersion prepared in S32 as the coating material, a bottom spray coating is performed in a fluidized bed coating machine to form hydrophobic microspheres. The hydrophobic microspheres are dispersed in water and dropped onto a silicon wafer. After slight centrifugation or natural sedimentation, the hydrophobic microspheres only contact the lower side of the substrate to form a monolayer distribution. Excess liquid is gently blown away and the microspheres are allowed to dry naturally in the air. A silicon-based hard mask is placed above the hydrophobic microsphere layer, ensuring that the mask holes exactly cover the microspheres. The hydrophobic microspheres are then treated with a plasma etching machine to obtain plasma-treated hydrophobic microspheres.

[0020] S34. Heat the phosphate buffer solution, weigh out inulin and gellan gum and dissolve them in the phosphate buffer solution. Let them stand to swell. After the gel solution cools down, mix Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus acidophilus evenly and add them to the gel solution. Then add skim milk powder and trehalose and stir to form a uniform bacterial gel suspension.

[0021] S35. Disperse the plasma-treated hydrophobic microspheres prepared in S33 in light mineral oil. Add the bacterial gel suspension prepared in S34 dropwise under slow stirring to form a water / oil type primary emulsion. Slowly drop the primary emulsion into a pre-cooled 2% CaCl2 solution, stir to crosslink and solidify. Collect the microcapsules through a 100-200 mesh standard sieve. Rinse the microcapsules several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, seal and store.

[0022] Preferably, in step S31, the mass ratio of food-grade glucose, food-grade sodium bicarbonate, and glucose oxidase is 35~50:20~30:1~4.

[0023] Preferably, in step S32, the mass ratio of carnauba wax, shea butter, and microcrystalline cellulose is 4~8:2~6:1~3.

[0024] Preferably, the amount of sodium stearoyl lactylate added in step S32 is 0.4 to 0.8 wt% of the total mass of carnauba wax and shea butter.

[0025] Preferably, in step S34, the mass ratio of inulin to gellan gum is 7~12:1~3.

[0026] Preferably, the mass concentration of hydrophobic microspheres in light mineral oil in step S35 is 5-15%.

[0027] Beneficial effects:

[0028] 1. The microcapsules prepared by this invention effectively block the stress of inhibitory components and high-viscosity polysaccharide environment in the *Hydrangea rubra* extract on *Lactobacillus* in the early stage of fermentation, creating a protected microenvironment for *Lactobacillus*, enabling it to preferentially utilize external nutrients for rapid proliferation, thus overcoming the bottlenecks of slow fermentation start-up and insufficient cell growth.

[0029] 2. This invention can trigger a gas-generating reaction during fermentation through enzymatic reactions within the microcapsules. Gas is directionally released from the micropores of the weakest hydrophobic layer or from channels disrupted by internal pressure, generating a reaction force that propels the microcapsules into random, autonomous movement within the *Hydrangea rubra* extract. This random, autonomous movement disrupts the high-viscosity liquid network created by *Hydrangea rubra* polysaccharides, improving the diffusion and mass transfer efficiency of O2, nutrients, and metabolites in the fermentation system, thereby significantly enhancing fermentation efficiency. Simultaneously, this continuous movement significantly increases the collision and contact opportunities between *Lactobacillus* and the macromolecular active substances of *Hydrangea rubra*, thus promoting the biodegradation and transformation of these substances by *Lactobacillus*, more fully releasing or generating new functional factors, and enhancing the core value of the product.

[0030] 3. The microcapsule provided by this invention serves as an independent and controllable micro-bioreactor. The gas production power and movement duration are precisely determined by the amount of pre-encapsulated core substrate. Its initiation depends on the metabolism of internal lactobacilli. This introduces a stable and predictable time control node for the fermentation process, fundamentally eliminating batch differences in traditional fermentation, and achieving precise control over the fermentation speed and endpoint, greatly ensuring the uniformity and stability of product quality.

[0031] 4. This invention overcomes the limitations of static physical isolation in traditional encapsulation technology, achieving dynamic intelligent protection. In the early stages of fermentation, the microcapsules remain stationary, protecting the lactobacilli from external environmental inhibition. In the later stages of fermentation, when the lactobacilli proliferate to a high density, dynamic movement is automatically triggered, actively improving external mass transfer conditions. This sequential design resolves the inherent contradiction in traditional methods where mass transfer efficiency is sacrificed to protect the cells, ensuring efficient fermentation while maintaining cell activity throughout the entire process. Attached Figure Description

[0032] Figure 1 A process flow diagram for fermented beverages made from Hydrangea macrophylla;

[0033] Figure 2 Photo of a fermented beverage made from Hydrangea macrophylla;

[0034] Figure 3 The viable cell counts of *Hydrangea macrophylla* fermented beverages in Examples 5 and 5-10;

[0035] Figure 4 The pH value and titratable acid content of *Hydrangea macrophylla* fermented beverages in Examples 5 and Comparative Examples 4-10;

[0036] Figure 5 The soluble solids content of the *Hydrangea macrophylla* fermented beverages in Examples 5 and Comparative Examples 4-10;

[0037] Figure 6 The soluble protein content of the *Hydrangea macrophylla* fermented beverages in Examples 5 and 4-10;

[0038] Figure 7 The free amino acid content of *Hydrangea macrophylla* fermented beverages in Example 5 and Comparative Examples 4-10;

[0039] Figure 8 The total phenolic content of the *Hydrangea macrophylla* fermented beverages in Examples 5 and 4-10;

[0040] Figure 9 The sensory evaluation results of the *Hydrangea macrophylla* fermented beverages of Example 5 and Comparative Examples 4-10 are as follows;

[0041] Figure 10 The results of electronic nose tests on the *Hydrangea macrophylla* fermented beverages of Example 5 and Comparative Examples 4-10 are shown.

[0042] Figure 11 O2 for the *Hydrangea macrophylla* fermented beverages of Examples 5 and 4-10 − • Free radical scavenging rate;

[0043] Figure 12 The ·OH radical scavenging rates of *Hydrangea macrophylla* fermented beverages in Example 5 and Comparative Examples 4-10 are shown.

[0044] Figure 13 The DPPH free radical scavenging rate of *Hydrangea macrophylla* fermented beverages in Example 5 and Comparative Examples 4-10;

[0045] Figure 14 The ABTS free radical scavenging rate of *Hydrangea macrophylla* fermented beverages in Example 5 and Comparative Examples 4-10 is given. Detailed Implementation

[0046] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments:

[0047] Glucose oxidase was purchased from Hebei Zhongzhisheng Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g; Lactobacillus paracasei ( L. paracasei Lactobacillus plantarum ( L. plantarum ), Lactobacillus acidophilus ( L. acidophilus Lactobacillus bulgaricus ( L. bulgaricus Bifidobacterium bifidum ( B.bifidum Lactobacillus rhamnosus ( L.rhamnosus All samples were purchased from Minsheng Zhongke Jiayi (Shandong) Biotechnology Co., Ltd., and the live bacteria count was 10 billion CFU / g.

[0048] Example 1

[0049] This embodiment describes a method for preparing microcapsules, including the following steps:

[0050] S1. Weigh 300 g of food-grade glucose, 200 g of food-grade sodium bicarbonate and 8 g of glucose oxidase. Mix them at 300 rpm for 20 min under ambient temperature of 25℃ and relative humidity of 30% to ensure that the materials are fully uniform. Then seal the mixed powder in a light-proof and moisture-proof container for later use.

[0051] S2. Weigh 175 g of carnauba wax and 50 g of shea butter, heat and melt them in a 90°C water bath, add 1.13 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 25 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a uniform hydrophobic wall material dispersion.

[0052] S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, a bottom-spray coating was performed in a fluidized bed coating machine. The inlet air temperature was controlled at 40℃, the atomization pressure at 1.5 Bar, the material temperature at 30℃, and the spray rate at 3 mL / min to form hydrophobic microspheres. The hydrophobic microspheres were then taken out and evenly spread in a sterile stainless steel tray and cooled to below 30℃. The hydrophobic microspheres were dispersed in deionized water at a concentration of 0.1% (w / v) and dropped onto a silicon wafer. The wafer was centrifuged or allowed to settle naturally so that the microspheres only contacted the lower side of the substrate, forming a monolayer distribution. Excess liquid was gently blown away, and the wafer was allowed to air dry naturally for 5 min. A silicon-based hard mask was placed above the microsphere layer, ensuring that the mask holes exactly covered the microspheres. A low-pressure oxygen plasma etching machine was used with the following settings: gas: O2, flow rate: 30 sccm, working pressure: 0.2 Pa, plasma power: 20 W, time: 35 s, to obtain plasma-treated hydrophobic microspheres.

[0053] S4. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 37.5 g of inulin and 7.5 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0054] S5. The hydrophobic microspheres prepared in S3 were dispersed in light mineral oil at 35℃ at a concentration of 10% (w / v). The bacterial gel suspension prepared in S4 was added dropwise under slow stirring at 250 rpm and emulsified for 25 min. Then, it was slowly added dropwise to a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stirred at 400 rpm for 30 min to fully crosslink and solidify. The microcapsules were collected through a 200-mesh standard sieve and rinsed several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, the microcapsules were sealed and stored.

[0055] Example 2

[0056] This embodiment describes a method for preparing microcapsules, including the following steps:

[0057] S1. Weigh 300 g of food-grade glucose, 200 g of food-grade sodium bicarbonate and 8 g of glucose oxidase. Mix them at 300 rpm for 20 min under ambient temperature of 25℃ and relative humidity of 30% to ensure that the materials are fully uniform. Then seal the mixed powder in a light-proof and moisture-proof container for later use.

[0058] S2. Weigh 139 g of carnauba wax and 83 g of shea butter, heat and melt them in a 90°C water bath, add 1.33 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 28 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a uniform hydrophobic wall material dispersion.

[0059] S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, a bottom-spray coating was performed in a fluidized bed coating machine. The inlet air temperature was controlled at 40℃, the atomization pressure at 1.5 Bar, the material temperature at 30℃, and the spray rate at 3 mL / min to form hydrophobic microspheres. The hydrophobic microspheres were then taken out and evenly spread in a sterile stainless steel tray and cooled to below 30℃. The hydrophobic microspheres were dispersed in deionized water at a concentration of 0.1% (w / v) and dropped onto a silicon wafer. The wafer was centrifuged or allowed to settle naturally so that the microspheres only contacted the lower side of the substrate, forming a monolayer distribution. Excess liquid was gently blown away, and the wafer was allowed to air dry naturally for 5 min. A silicon-based hard mask was placed above the microsphere layer, ensuring that the mask holes exactly covered the microspheres. A low-pressure oxygen plasma etching machine was used with the following settings: gas: O2, flow rate: 30 sccm, working pressure: 0.2 Pa, plasma power: 20 W, time: 35 s, to obtain plasma-treated hydrophobic microspheres.

[0060] S4. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 37.5 g of inulin and 7.5 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0061] S5. The hydrophobic microspheres prepared in S3 were dispersed in light mineral oil at 35℃ at a concentration of 10% (w / v). The bacterial gel suspension prepared in S4 was added dropwise under slow stirring at 250 rpm and emulsified for 25 min. Then, it was slowly added dropwise to a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stirred at 400 rpm for 30 min to fully crosslink and solidify. The microcapsules were collected through a 200-mesh standard sieve and rinsed several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, the microcapsules were sealed and stored.

[0062] Example 3

[0063] This embodiment describes a method for preparing microcapsules, including the following steps:

[0064] S1. Weigh 300 g of food-grade glucose, 200 g of food-grade sodium bicarbonate and 8 g of glucose oxidase. Mix them at 300 rpm for 20 min under ambient temperature of 25℃ and relative humidity of 30% to ensure that the materials are fully uniform. Then seal the mixed powder in a light-proof and moisture-proof container for later use.

[0065] S2. Weigh 175 g of carnauba wax and 50 g of shea butter, heat and melt them in a 90°C water bath, add 0.9 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 25 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a uniform hydrophobic wall material dispersion.

[0066] S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, a bottom-spray coating was performed in a fluidized bed coating machine. The inlet air temperature was controlled at 40℃, the atomization pressure at 1.5 Bar, the material temperature at 30℃, and the spray rate at 3 mL / min to form hydrophobic microspheres. The hydrophobic microspheres were then taken out and evenly spread in a sterile stainless steel tray and cooled to below 30℃. The hydrophobic microspheres were dispersed in deionized water at a concentration of 0.1% (w / v) and dropped onto a silicon wafer. The wafer was centrifuged or allowed to settle naturally so that the microspheres only contacted the lower side of the substrate, forming a monolayer distribution. Excess liquid was gently blown away, and the wafer was allowed to air dry naturally for 5 min. A silicon-based hard mask was placed above the microsphere layer, ensuring that the mask holes exactly covered the microspheres. A low-pressure oxygen plasma etching machine was used with the following settings: gas: O2, flow rate: 30 sccm, working pressure: 0.2 Pa, plasma power: 20 W, time: 35 s, to obtain plasma-treated hydrophobic microspheres.

[0067] S4. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 35 g of inulin and 10 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0068] S5. The hydrophobic microspheres prepared in S3 were dispersed in light mineral oil at 35℃ at a concentration of 10% (w / v). The bacterial gel suspension prepared in S4 was added dropwise under slow stirring at 250 rpm and emulsified for 25 min. Then, it was slowly added dropwise to a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stirred at 400 rpm for 30 min to fully crosslink and solidify. The microcapsules were collected through a 200-mesh standard sieve and rinsed several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, the microcapsules were sealed and stored.

[0069] Example 4

[0070] This embodiment describes a method for preparing microcapsules, including the following steps:

[0071] S1. Weigh 300 g of food-grade glucose, 200 g of food-grade sodium bicarbonate and 8 g of glucose oxidase. Mix them at 300 rpm for 20 min under ambient temperature of 25℃ and relative humidity of 30% to ensure that the materials are fully uniform. Then seal the mixed powder in a light-proof and moisture-proof container for later use.

[0072] S2. Weigh 150 g of carnauba wax and 50 g of shea butter, heat and melt them in a 90°C water bath, add 1.0 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 50 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a uniform hydrophobic wall material dispersion.

[0073] S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, a bottom-spray coating was performed in a fluidized bed coating machine. The inlet air temperature was controlled at 40℃, the atomization pressure at 1.5 Bar, the material temperature at 30℃, and the spray rate at 3 mL / min to form hydrophobic microspheres. The hydrophobic microspheres were then taken out and evenly spread in a sterile stainless steel tray and cooled to below 30℃. The hydrophobic microspheres were dispersed in deionized water at a concentration of 0.1% (w / v) and dropped onto a silicon wafer. The wafer was centrifuged or allowed to settle naturally so that the microspheres only contacted the lower side of the substrate, forming a monolayer distribution. Excess liquid was gently blown away, and the wafer was allowed to air dry naturally for 5 min. A silicon-based hard mask was placed above the microsphere layer, ensuring that the mask holes exactly covered the microspheres. A low-pressure oxygen plasma etching machine was used with the following settings: gas: O2, flow rate: 30 sccm, working pressure: 0.2 Pa, plasma power: 20 W, time: 35 s, to obtain plasma-treated hydrophobic microspheres.

[0074] S4. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 41 g of inulin and 4 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0075] S5. The hydrophobic microspheres prepared in S3 were dispersed in light mineral oil at 35℃ at a concentration of 5% (w / v). The bacterial gel suspension prepared in S4 was added dropwise under slow stirring at 250 rpm and emulsified for 25 min. Then, it was slowly added dropwise to a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stirred at 400 rpm for 30 min to fully crosslink and solidify. The microcapsules were collected through a 200-mesh standard sieve and rinsed several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, the microcapsules were sealed and stored.

[0076] Table 1 Encapsulation efficiency of microcapsules

[0077]

[0078] As shown in Table 1, the encapsulation rates of the microcapsules prepared in Examples 1-4 for the complex lactobacilli (Lactobacillus paracasei, Lactobacillus plantarum, and Lactobacillus acidophilus) were all above 85%, indicating that the method provided by this invention can achieve efficient encapsulation of complex lactobacilli under different process parameters.

[0079] Example 5

[0080] This embodiment describes a method for preparing a fermented beverage made from *Hydrangea macrophylla*, including the following steps:

[0081] S1. Select fresh, non-rotten *Hydrangea macrophylla*, wash and tear into 1 cm pieces, add to 100℃ hot water at a solid-liquid ratio of 1:10, extract for 60 min, filter the extract through a 400-mesh filter cloth to obtain *Hydrangea macrophylla* extract.

[0082] S2. Add 7% of the mass of the hydrangea extract to white sugar, and pasteurize in a constant temperature magnetic stirring water bath at 85°C for 15 min. After sterilization, transfer the extract to a sterile fermenter in a clean bench and cool to room temperature for later use.

[0083] S3. Under aseptic conditions, add 2% by mass of the extract of the microcapsules prepared in Example 1 to the fermenter and let the microcapsules ferment at a constant temperature of 37°C for 36 h.

[0084] S4. After fermentation, sterilize in a constant temperature magnetic stirring water bath at 85℃ for 15 minutes, then hot fill and cap. For industrial production with a long shelf life, UHT sterilization for about 5 seconds can be performed first, followed by sterilization at 85~90℃ for 5~10 minutes after filling.

[0085] Example 6.

[0086] The difference between this embodiment and embodiment 5 is that in this embodiment, the amount of microcapsules added is 0.8% of the mass of the extract, and the remaining steps are the same as in embodiment 5.

[0087] Example 7

[0088] The difference between this embodiment and embodiment 5 is that in this embodiment, the amount of microcapsules added is 1.5% of the mass of the extract, and the remaining steps are the same as in embodiment 5.

[0089] Example 8

[0090] The difference between this embodiment and embodiment 5 is that in this embodiment, the amount of microcapsules added is 2.5% of the mass of the extract, and the remaining steps are the same as in embodiment 5.

[0091] Example 9

[0092] This embodiment describes a method for preparing a fermented beverage made from *Hydrangea macrophylla*, including the following steps:

[0093] S1. Select fresh, non-rotten *Hydrangea macrophylla*, wash and tear into 1 cm pieces, add 95℃ hot water at a solid-liquid ratio of 1:4, extract for 60 min, filter the extract through a 400-mesh filter cloth to obtain *Hydrangea macrophylla* extract.

[0094] S2. Add 12% of the mass of the hydrangea extract to white sugar, and pasteurize it in a constant temperature magnetic stirring water bath at 85°C for 15 min. After sterilization, transfer the extract to a sterile fermenter in a clean bench and cool it to room temperature for later use.

[0095] S3. Under aseptic conditions, add 2% by weight of the extract of the microcapsules prepared in Example 1 to the fermenter and let it ferment at a constant temperature of 36°C for 40 h.

[0096] S4. After fermentation, sterilize in a constant temperature magnetic stirring water bath at 85℃ for 15 minutes, then fill and seal.

[0097] Example 10

[0098] This embodiment describes a method for preparing a fermented beverage made from *Hydrangea macrophylla*, including the following steps:

[0099] S1. Select fresh, non-rotten *Hydrangea macrophylla*, wash and tear into 1 cm pieces, add to 90℃ hot water at a solid-liquid ratio of 1:8, extract for 60 min, filter the extract through a 400-mesh filter cloth to obtain *Hydrangea macrophylla* extract.

[0100] S2. Add 8% of the mass of the hydrangea extract to white sugar, and pasteurize in a constant temperature magnetic stirring water bath at 85°C for 15 min. After sterilization, transfer the extract to a sterile fermenter in a clean bench and cool to room temperature for later use.

[0101] S3. Under aseptic conditions, add 2% by weight of the extract of the microcapsules prepared in Example 1 to the fermenter and let it ferment at a constant temperature of 37°C for 45 h.

[0102] S4. After fermentation, sterilize in a constant temperature magnetic stirring water bath at 85℃ for 15 minutes, then fill and seal.

[0103] Example 11

[0104] This embodiment describes a method for preparing a fermented beverage made from *Hydrangea macrophylla*, including the following steps:

[0105] S1. Select fresh, unrotted *Hydrangea macrophylla*, wash and tear into 1 cm pieces, add 95°C hot water at a solid-liquid ratio of 1:10, extract for 60 min, filter the extract through a 400-mesh filter cloth to obtain *Hydrangea macrophylla* extract.

[0106] S2. Add 15% of the mass of the hydrangea extract to white sugar, and pasteurize in a constant temperature magnetic stirring water bath at 85°C for 15 min. After sterilization, transfer the extract to a sterile fermenter in a clean bench and cool to room temperature for later use.

[0107] S3. Under aseptic conditions, 2% by weight of the extract of the microcapsules prepared in Example 1 was added to the fermenter and fermented at a constant temperature of 38°C for 36 h.

[0108] S4. After fermentation, sterilize in a constant temperature magnetic stirring water bath at 85℃ for 15 minutes, then fill and seal.

[0109] Comparative Example 1

[0110] The difference between this comparative example and Example 5 is that in this comparative example, unencapsulated compound lactobacillus is directly added. The compound lactobacillus is a premix of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus acidophilus in a mass ratio of 1:1:1. The remaining steps are the same as in Example 5.

[0111] Comparative Example 2

[0112] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example do not contain a core composed of food-grade glucose, food-grade sodium bicarbonate, and glucose oxidase, and include the following steps:

[0113] S1. Weigh 175 g of carnauba wax and 50 g of shea butter, heat and melt them in a 90°C water bath, add 1.13 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 25 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a homogeneous oil phase;

[0114] S2. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 37.5 g of inulin and 7.5 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0115] S3. Stir the bacterial gel suspension and oil phase evenly and emulsify for 25 min to form a water / oil type pre-emulsion. Slowly drip the pre-emulsion into a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stir at 400 rpm for 30 min to fully cross-link and solidify. Collect the microcapsules through a 200-mesh standard sieve. Rinse the microcapsules several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, seal and store.

[0116] S4. The preparation method of the *Hydrangea macrophylla* fermented beverage is the same as in Example 5.

[0117] Comparative Example 3

[0118] The difference between this comparative example and Example 5 is that this comparative example does not involve plasma treatment of the hydrophobic microspheres, but includes the following steps:

[0119] S1. Weigh 300 g of food-grade glucose, 200 g of food-grade sodium bicarbonate and 8 g of glucose oxidase. Mix them at 300 rpm for 20 min under ambient temperature of 25℃ and relative humidity of 30% to ensure that the materials are fully uniform. Then seal the mixed powder in a light-proof and moisture-proof container for later use.

[0120] S2. Weigh 175 g of carnauba wax and 50 g of shea butter, heat and melt them in a 90°C water bath, add 1.13 g of sodium stearoyl lactylate, stir to form a homogeneous oil phase, lower the temperature to 55°C, add 25 g of microcrystalline cellulose, stir at 150 rpm for 10 min, increase the speed to 1000 rpm and stir for 45 min to form a uniform hydrophobic wall material dispersion.

[0121] S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, perform bottom spray coating in a fluidized bed coating machine. Control the inlet air temperature at 40℃, the atomization pressure at 1.5 Bar, the material temperature at 30℃, and the spray rate at 3 mL / min to form hydrophobic microspheres. Take out the hydrophobic microspheres and spread them evenly in a sterile stainless steel tray, and cool them to below 30℃.

[0122] S4. Heat 1000 mL of phosphate buffer (0.05 mol / L, pH 6.8) to 50°C. Weigh 37.5 g of inulin and 7.5 g of gellan gum and dissolve them in the phosphate buffer. Let the solution swell for 1.5 h. After the gel solution cools to 35°C, mix 16 g of skim milk powder, 4 g of trehalose, 1 g of Lactobacillus paracasei, 1 g of Lactobacillus plantarum and 1 g of Lactobacillus acidophilus evenly and add them to the gel solution. Stir at 80 rpm for 20 min to form a homogeneous bacterial gel suspension.

[0123] S5. The hydrophobic microspheres prepared in S3 were dispersed in light mineral oil at 35℃ at a concentration of 10% (w / v). The bacterial gel suspension prepared in S4 was added dropwise under slow stirring at 250 rpm and emulsified for 25 min to form a water / oil type primary emulsion. The primary emulsion was slowly added dropwise to a pre-cooled (4℃) 2% (w / v) CaCl2 solution and stirred at 400 rpm for 30 min to fully crosslink and solidify. The microcapsules were collected through a 200-mesh standard sieve and rinsed several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, the microcapsules were sealed and stored.

[0124] S6. The preparation method of the *Hydrangea macrophylla* fermented beverage is the same as in Example 5.

[0125] Table 2 Physicochemical indicators of *Hydrangea macrophylla* fermentation broth

[0126]

[0127] As shown in Table 2, compared with Comparative Example 1 (unencapsulated Lactobacillus complex), the viable cell count in the *Hydrangea macrophylla* fermentation broth of Examples 5-11 (microcapsules) was lower during the 0-4 h period. This indicates that the hydrophobic wall material (carnauba wax, shea butter microcrystalline cellulose) effectively blocked the contact between the internal bacteria and the inhibitors in the external *Hydrangea macrophylla* extract. At this time, all the bacterial agents were in a sedimentation state, indicating that the gas-producing core of the microcapsule had not yet been activated. The viable cell counts of Comparative Example 2 (without glucose oxidase core) and Comparative Example 3 (encapsulation structure defect) were between those of Examples 5-11 and Comparative Example 1, indicating that although simple encapsulation (Comparative Example 2) or imperfect encapsulation (Comparative Example 3) had a certain protective effect on the bacteria, the effect was not as good as that of Examples 5-11. In the early stage of fermentation, water and nutrients in the *Hydrangea macrophylla* extract permeated into the microcapsules through the hydrophilic loose layer (inulin-gellan gum layer), providing nutrients for the internal Lactobacillus. The Lactobacillus began to proliferate and produce acid. When fermentation progressed to 4-10 h... h. In Examples 5-11, the bacterial agent changed from sedimentation to suspension. This is because the microenvironment inside the microcapsule gradually erodes the dense hydrophobic layer due to lactic acid accumulation and the lactobacillus' own lipase, while enhancing the swelling capacity of the loose hydrophilic layer. Water enters the core, activating glucose oxidase, initiating the enzymatic reaction, and catalyzing the conversion of glucose inside the microcapsule into gluconic acid and H2O2. At this time, because the bacterial cells are already in a high-density state, their abundant H2O2 system can immediately decompose the H2O2 diffused to the gel layer into water and O2, while gluconic acid undergoes a neutralization reaction with sodium bicarbonate. The continuous generation of CO2 gas creates high pressure within the cavity, eventually leading to its directional discharge from the weakest point of the hydrophobic layer (micropores) or channels disrupted by internal pressure. This generates a reaction force that propels the microcapsules to move autonomously within the *Hydrangea rubra* extract. The internally generated gas, produced and released from within the microcapsules, provides initial momentum. This continuous movement and efficient mass transfer increase the collision frequency and contact opportunities between the encapsulated *Lactobacillus* and the macromolecules of *Hydrangea rubra* in the extract, thereby promoting the degradation and transformation of these macromolecules by the *Lactobacillus* enzyme system and enhancing the product's functionality. In contrast, none of the comparative examples achieved this synergistic effect: Comparative Example 1, lacking encapsulation protection, suffered from impaired bacterial activity; Comparative Example 2, lacking a gas-generating core, showed an increase in viable bacteria count but constant sedimentation of the bacterial agent; Comparative Example 3, due to defects in its encapsulation structure, behaved more like Comparative Example 2 and also failed to achieve autonomous movement and controlled release.

[0128] Comparative Example 4

[0129] This comparative example is the extract of *Hydrangea macrophylla* without the addition of inoculants.

[0130] Comparative Example 5

[0131] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Lactobacillus plantarum (LPT), and the specific process is the same as in Example 5.

[0132] Comparative Example 6

[0133] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Lactobacillus paracasei (LPC), and the specific process is the same as in Example 5.

[0134] Comparative Example 7

[0135] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Lactobacillus acidophilus (LA), and the specific process is the same as in Example 5.

[0136] Comparative Example 8

[0137] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Lactobacillus rhamnosus (LR), and the specific process is the same as in Example 5.

[0138] Comparative Example 9

[0139] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Bifidobacterium bifidum (BB), and the specific process is the same as in Example 5.

[0140] Comparative Example 10

[0141] The difference between this comparative example and Example 5 is that the microcapsules in this comparative example are replaced with unencapsulated Lactobacillus bulgaricus (LB), and the specific process is the same as in Example 5.

[0142] Table 3 Color difference of *Hydrangea macrophylla* fermented beverage

[0143]

[0144] As shown in Table 3, Comparative Example 4 has the highest L* value, indicating that the unfermented extract itself is brighter. Example 5 has a lower L* value, and the L* values ​​of Comparative Examples 5-10 are all lower than those of Comparative Example 4, indicating that lactic acid bacteria fermentation generally reduces brightness. Comparative Example 10 (unencapsulated Lactobacillus bulgaricus) has a relatively high L* value, while Example 5 and Comparative Examples 5-7 have lower L* values. Reasons: (1) Organic acids (such as lactic acid) are produced during lactic acid bacteria fermentation, which lowers pH and may promote Maillard reaction or enzymatic browning. These reactions produce brown substances, thereby reducing the L* value. Example 5 and Comparative Examples 5-7, which use a single Lactobacillus, fermented more thoroughly, resulting in a more significant decrease in brightness; (2) Lactobacillus bulgaricus (Comparative Example 10) may have a milder metabolism, producing less acid or enzymes, resulting in a lower degree of browning and therefore a higher L* value. Microcapsules (Example 5) make fermentation more uniform and slower, promote pigment degradation, and further reduce brightness; (3) Encapsulation protects the cells, prolongs fermentation time, and may enhance the release of metabolites, leading to more obvious browning. The fermentation of unencapsulated strains (Comparative Examples 5-10) may be faster but not thorough, so the L* values ​​vary. Lactic acid bacteria fermentation may degrade chlorophyll, resulting in a weakening of green. The a* values ​​of Example 5 and Comparative Examples 5, 6, 7, and 9 are closer to 0 than the blank control, indicating that fermentation reduces green. It is also possible that different lactic acid bacteria produce different enzymes (such as chlorophyllase) or metabolites, affecting pigment stability. Comparative Example 10 may produce fewer enzymes that degrade chlorophyll, or secrete substances to protect green pigment, so the a* value is more negative. Encapsulation may control the metabolic rate and reduce the drastic degradation of pigment, so the b* value of Example 5 is similar to that of Comparative Examples 5-7. Example 5 balances metabolism and avoids excessive degradation. Comparative Example 10 may have higher oxidative activity, accelerating the degradation of yellow pigment, so the b* value is the lowest. Comparative Examples 5 and 6 showed weaker degradation ability and higher b values.

[0145] like Figure 3As shown, Example 5 had the highest viable count, possibly because the encapsulation carrier (such as sodium alginate) constructed a "microecological barrier" for the lactic acid bacteria, buffering the stress on the strains caused by pH decrease and metabolic product accumulation during the fermentation of *Lactobacillus acidophilus* extract, reducing damage to viable bacteria, and improving the survival and proliferation capacity of the strains. The synergistic growth of the complex bacteria: the metabolic products of different lactic acid bacteria (such as organic acids and extracellular polysaccharides) can be mutually utilized, creating a more suitable growth environment for each other, further improving the viable bacteria retention rate. Comparative Example 7 and Example 5 showed no significant difference, possibly because *Lactobacillus acidophilus* itself has strong resistance and can adapt well to the fermentation environment of *Lactobacillus acidophilus* extract even without encapsulation. Its own metabolic characteristics (such as mild acid production and high nutrient utilization efficiency) are well-suited to the extract, thus achieving a viable count comparable to that of the encapsulated complex bacteria. The reason why Comparative Example 9 showed no difference from Example 5 is that the growth characteristics of *Bifidobacterium bifidum* are highly compatible with the *Hydrangea breviscapus* extract. The nutrients in the extract (such as polysaccharides and small peptides) can meet its proliferation requirements. Furthermore, this strain has strong tolerance to environmental stress and can maintain a high viable cell retention rate during fermentation even without encapsulation. Comparative Example 10 had the lowest viable cell count, possibly because the optimal growth conditions for *Lactobacillus bulgaricus* (such as nutritional requirements and pH tolerance range) did not match the environment of the *Hydrangea breviscapus* extract. This made the strain susceptible to environmental stress, ultimately resulting in weak proliferation and the lowest viable cell count.

[0146] like Figure 4 As shown, the microcapsules prepared by this invention (Example 5) performed best—ensuring the activity of the strains. The synergistic metabolism of the compound bacteria not only improved the survival rate of live bacteria but also made full use of the nutrients in the extract to produce a large amount of acid (low pH, high TA), ultimately achieving a better sensory experience. In contrast, Comparative Example 4, which did not add bacterial powder, had a higher pH and extremely low TA due to the lack of fermentation. Comparative Examples 5 to 10, due to the lack of encapsulation protection and differences in strain characteristics (stress resistance, acid production capacity) and compatibility with the extract, showed varying degrees of fluctuation in live bacteria count, pH / TA, and sensory scores. Among them, some strains with strong stress resistance (such as Lactobacillus rhamnosus) could achieve a higher live bacteria count, but their fermentation acid production and sensory performance were still weaker than those of the encapsulated compound bacteria group.

[0147] like Figure 5 As shown, Comparative Example 4 had the highest TSS level because no fermentation process was involved, and the soluble substances in the extract were not metabolized and utilized. Example 5 had the lowest TSS level because the encapsulation technology ensured the activity of the strains, and the synergistic metabolism of the compound bacteria efficiently consumed the soluble substances (such as sugars and polysaccharides) in the extract. Comparative Examples 5-10 showed fluctuations in TSS due to differences in the metabolic capacity of the strains and their compatibility with the extracts. Strains with strong metabolic capacity (such as Lactobacillus plantarum) utilized soluble substances more fully and had lower TSS levels, while strains with poor compatibility (such as Lactobacillus bulgaricus) had weak metabolic efficiency and relatively higher TSS levels.

[0148] like Figure 6As shown, Comparative Example 10 had the highest soluble protein content due to the high protease activity and low protein consumption of the strain; Example 5 had a relatively high soluble protein content due to the synergistic decomposition of protein by the complex bacterial enzyme system; Comparative Example 4 had the lowest soluble protein content because there was no fermentation and no protease activity; and the soluble protein content of the unencapsulated single bacterial group fluctuated due to the differences in protease activity and metabolic consumption of the strains.

[0149] like Figure 7 As shown, the activity of the encapsulated complex bacteria in Example 5 is stable, and the proteases and peptidases secreted by different strains work synergistically to efficiently decompose proteins and peptides in the extract, generating a large amount of free amino acids. At the same time, the metabolic consumption of the complex bacteria is less than the decomposition efficiency, so the content of free amino acids is the highest. Although Comparative Example 7 has a certain protease activity, the consumption of free amino acids by the growth of the strain is much greater than the amount generated by its decomposition, resulting in its free amino acid content being even lower than that of the non-fermented Comparative Example 4.

[0150] like Figure 8 As shown, in Example 5, the activity was stable under encapsulation protection, and the metabolic process of the compound bacteria promoted the release / conversion of phenolic substances in the *Hydrangea macrophylla* extract: on the one hand, enzymes secreted by the strain (such as glycosidases) could decompose phenolic compounds (such as phenolic glycosides) in the extract, converting them into free phenols; on the other hand, the synergistic metabolism of the compound bacteria created a suitable environment, reducing the oxidative loss of phenolic substances, thus resulting in the highest total phenol content. In contrast, Example 4 had the lowest total phenol content due to the lack of enzymatic hydrolysis and natural loss of phenols; the total phenol content of the unencapsulated single-strain group fluctuated due to differences in the enzymatic hydrolysis ability of the strains and the consumption / degradation of phenols.

[0151] like Figure 9As shown, Example 5 received the highest score, possibly because the encapsulation protected the activity of the lactic acid bacteria, resulting in stronger strain stability and more sustained metabolism during fermentation. This allowed for more efficient utilization of the nutrients in the *Hydrangea macrophylla* extract, generating more metabolites that improve flavor and texture (such as organic acids and esters). The complementary metabolic pathways of different lactic acid bacteria not only adjusted the acidity of the extract but also enriched the flavor profile, ultimately leading to better sensory performance. Comparative Example 4 received the lowest score because the flavor of the *Hydrangea macrophylla* extract itself (such as the raw or fishy taste of the raw materials) was not improved, and there were no pleasant flavor compounds produced by lactic acid bacteria metabolism, resulting in the worst sensory experience. The scores of Comparative Examples 5-10 were lower than those of Example 5, possibly because the lack of encapsulation made the lactic acid bacteria more susceptible to environmental factors (such as the pH and osmotic pressure of the *Lactobacillus caudatus* extract) during fermentation, resulting in rapid loss of activity, low fermentation efficiency, insufficient metabolites, and weaker flavor / texture improvement compared to the encapsulated group. Furthermore, the flavor profile of single-strain fermentation was thin: the metabolic capacity of different single strains was limited (for example, some strains produced strong acid but had a monotonous flavor, while others had a weak flavor), making it impossible to synergistically optimize the sensory experience like a complex strain. Additionally, the fermentation characteristics of some strains (such as *Lactobacillus bulgaricus* in Comparative Example 10) were poorly compatible with the *Lactobacillus caudatus* extract, potentially producing undesirable flavors (such as excessive acidity or bitterness) after single fermentation, thus resulting in lower scores than Example 5.

[0152] like Figure 10 As shown, Example 5 exhibits significantly higher response values ​​at sensors S1, S20, and S28, indicating a richer variety and higher content of flavor compounds. The synergistic fermentation of the encapsulated complex bacteria generates a more diverse range of metabolites (such as organic acids, esters, and aldehydes), while the encapsulation technology ensures strain activity and a more complete fermentation process, resulting in a fuller flavor profile. Comparative Example 4 shows generally lower and more concentrated sensor responses, indicating a lack of flavor compounds. Without a fermentation process, the *Hydrangea macrophylla* extract only retains the basic flavor of the raw material, lacking the characteristic flavor compounds produced by lactic acid bacteria metabolism. The flavor responses of some single bacteria (such as Comparative Examples 5 and 6) are close to those of Example 5, but the overall intensity is weaker. The fermentation of these strains can produce some characteristic flavors, but due to the lack of encapsulation protection and synergistic effect, the variety and content of flavor compounds are weaker than those of the encapsulated complex bacteria.

[0153] like Figure 11As shown, Example 5 exhibited the highest scavenging rate and stable activity under encapsulation protection, with synergistic fermentation generating more antioxidants: the metabolites of the complex bacteria (such as phenols, organic acids, and extracellular polysaccharides) possess strong antioxidant properties; the encapsulation technology enabled the strains to fully utilize the nutrients in the *Hydrangea macrophylla* extract (such as polysaccharides and polyphenol precursors), converting them into more antioxidant active substances, thus resulting in the highest superoxide anion scavenging rate. The reason for the extremely low scavenging rate in Comparative Example 4 is that there was no fermentation process; it relied solely on the basic antioxidants in the *Hydrangea macrophylla* extract itself, lacking additional antioxidant components produced by lactic acid bacteria metabolism, thus its scavenging rate was significantly lower than the fermentation group. Comparative Examples 6 and 10 (*Lactobacillus paracasei* and *Lactobacillus bulgaricus*): the strains produced fewer antioxidants through metabolism and consumed more of the original antioxidant components in the extract, resulting in lower scavenging rates.

[0154] like Figure 12 As shown, in Example 5, the activity was stable under encapsulation protection, and the synergistic fermentation enriched more antioxidant active substances: the metabolites of the complex bacteria (such as phenols, extracellular polysaccharides, and small molecule peptides) synergistically enhanced the scavenging ability of •OH by working with the original components of the *Hydrangea macrophylla* extract (such as polysaccharides); at the same time, the encapsulation technology ensured the full metabolism of the strain, further increasing the amount of antioxidant substances produced, thus the scavenging rate was at a high level. The reason for the low scavenging rate of Comparative Example 4 may be because there was no fermentation process, and it relied only on the basic antioxidant components of the *Hydrangea macrophylla* extract itself, without additional antioxidant substances produced by lactic acid bacteria metabolism, so the •OH scavenging rate was significantly lower than that of most fermentation groups. The antioxidant substances (such as organic acids and peptides) produced by the fermentation of Comparative Examples 6 and 8 were well compatible with the components of the extract, so the scavenging rate was close to that of Example 5; the antioxidant substances produced by the metabolism of Comparative Example 10 were few, and the consumption / destruction effect on the original antioxidant components of the extract was strong, so the scavenging rate was the lowest.

[0155] like Figure 13As shown, the co-fermentation in the examples generated a richer amount of antioxidants: the metabolites of the complex bacteria (such as phenols, organic acids, and extracellular polysaccharides) and the original components of the *Hydrangea macrophylla* extract (such as polysaccharides) formed a synergistic antioxidant effect. Simultaneously, the encapsulation technology allowed the strain to fully utilize the nutrients in the extract, further increasing the amount of antioxidants produced. Therefore, the DPPH scavenging rate was significantly higher than in other groups. *Lactobacillus acidophilus* (Comparative Example 7) can secrete enzymes such as proteases and glycosidases to break down macromolecules (such as proteins and phenolic glycosides) in the *Hydrangea macrophylla* extract into smaller active ingredients (such as antioxidant peptides and free phenols). At the same time, the strain's own metabolism can produce extracellular polysaccharides, short-chain fatty acids, and other antioxidants, which can scavenge DPPH free radicals through hydrogen atom transfer and electron transfer. The reason why the DPPH scavenging rate of Comparative Example 10 was lower than that of the unfermented group is twofold: Firstly, this strain has a stronger utilization / degradation effect on the original antioxidant components (such as total phenols and anthocyanins) in the *Hydrangea macrophylla* extract—studies have found that some lactic acid bacteria (including *Lactobacillus bulgaricus*) consume antioxidants such as phenols and anthocyanins when fermenting plant substrates; secondly, this strain generated less antioxidants in this system, which could not compensate for the loss of the original components. The reason why Comparative Example 4 had a low scavenging rate is that it did not undergo fermentation and relied solely on the basic antioxidant components of the *Hydrangea macrophylla* extract itself, without additional antioxidants produced by lactic acid bacteria metabolism. Therefore, its DPPH scavenging rate was much lower than that of the fermented group.

[0156] like Figure 14 As shown, the ABTS scavenging rate decreased significantly after fermentation. This may be because the natural antioxidants (such as phenols, polysaccharides, and flavonoids) in the *Hydrangea macrophylla* extract are the core substances for ABTS scavenging. However, during lactic acid bacteria fermentation, the strain consumes these components through enzymatic hydrolysis and metabolism. Glycosidases and esterases secreted by some lactic acid bacteria (such as *Lactobacillus plantarum* and *Lactobacillus bulgaricus*) degrade phenolic substances (such as phenolic glycosides), converting them into small molecules with no antioxidant activity. The antioxidant activity of the newly generated substances (such as short-chain fatty acids) is weaker than that of the original natural components and cannot equivalently replace their ABTS scavenging ability. The reason for the largest decrease in ABTS scavenging rate in Comparative Example 10 is twofold: firstly, the enzymes secreted by this strain (such as phenolic esterases) degrade a large amount of the core antioxidant components such as phenols and flavonoids in the extract; secondly, its compatibility with the *Hydrangea macrophylla* extract is poor, and the content of active substances such as extracellular polysaccharides and antioxidant peptides generated during growth and metabolism is extremely low, unable to compensate for the loss of the original components, ultimately leading to the largest decrease in ABTS scavenging rate.

[0157] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A fermented beverage made from *Hydrangea macrophylla*, characterized in that: The microcapsules were prepared by fermenting *Hydrangea macrophylla* extract using microcapsules. The microcapsules have an asymmetric structure: one side is a dense hydrophobic layer composed of carnauba wax, shea butter, and microcrystalline cellulose, containing glucose, sodium bicarbonate, and glucose oxidase; the other side is a loose hydrophilic layer composed of inulin and gellan gum composite gel, loaded with a composite of *Lactobacillus*. The preparation method of the microcapsules includes the following steps: S1. Weigh out food-grade glucose, food-grade sodium bicarbonate and glucose oxidase, mix them evenly, and seal the mixed powder in a light-proof and moisture-proof container for later use. S2. Weigh out carnauba wax and shea butter, heat them in a water bath to melt them, add sodium stearoyl lactylate, stir to form a homogeneous oil phase, then add microcrystalline cellulose and continue stirring to form a uniform hydrophobic wall material dispersion. S3. Using the mixed powder prepared in S1 as the core, and the hydrophobic wall material dispersion prepared in S2 as the coating material, a bottom-spray coating is performed in a fluidized bed coating machine to form hydrophobic microspheres. The hydrophobic microspheres are dispersed in water and dropped onto a silicon wafer. After slight centrifugation or natural sedimentation, the hydrophobic microspheres only contact the lower side of the substrate to form a monolayer distribution. Excess liquid is gently blown away and the microspheres are allowed to dry naturally in the air. A silicon-based hard mask is placed above the hydrophobic microsphere layer, ensuring that the mask holes exactly cover the microspheres. The hydrophobic microspheres are then treated with a plasma etching machine to obtain plasma-treated hydrophobic microspheres. S4. Heat the phosphate buffer solution, weigh out inulin and gellan gum and dissolve them in the phosphate buffer solution. Let them stand to swell. After the gel solution cools down, mix Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus acidophilus evenly and add them to the gel solution. Then add skim milk powder and trehalose and stir to form a homogeneous bacterial gel suspension. S5. Disperse the plasma-treated hydrophobic microspheres prepared in S3 in light mineral oil. Add the bacterial gel suspension prepared in S4 dropwise under slow stirring to form a water / oil type primary emulsion. Slowly drop the primary emulsion into a pre-cooled 2% CaCl2 solution, stir to crosslink and solidify. Collect the microcapsules through a 100-200 mesh standard sieve. Rinse the microcapsules several times with sterile water containing 0.1% food-grade Tween 80 and sterile water. After vacuum freeze-drying, seal and store.

2. The *Hydrangea macrophylla* fermented beverage according to claim 1, characterized in that: In step S1, the mass ratio of food-grade glucose, food-grade sodium bicarbonate, and glucose oxidase is 35~50:20~30:1~4.

3. The *Hydrangea macrophylla* fermented beverage according to claim 1, characterized in that: In step S2, the mass ratio of carnauba wax, shea butter, and microcrystalline cellulose is 4~8:2~6:1~3; the amount of sodium stearoyl lactylate added is 0.4~0.8 wt% of the total mass of carnauba wax and shea butter.

4. The *Hydrangea macrophylla* fermented beverage according to claim 1, characterized in that: In step S4, the mass ratio of inulin to gellan gum is 7~12:1~3.

5. A *Hydrangea macrophylla* fermented beverage according to claim 1, characterized in that: In step S5, the mass concentration of hydrophobic microspheres in light mineral oil is 5-15%.

6. A method for preparing a *Hydrangea macrophylla* fermented beverage according to any one of claims 1 to 5, characterized in that, Includes the following steps: (1) Select fresh, unrotten *Hydrangea macrophylla*, wash and tear into 1-3 cm pieces, add to water at 90-95℃ at a solid-liquid ratio of 1:4-10, extract for 60 min, filter with 300-400 mesh filter cloth to obtain *Hydrangea macrophylla* extract. (2) Add 7-15% of the mass of the hydrangea extract with white sugar, pasteurize in a constant temperature magnetic stirring water bath at 80-85℃ for 10-15 min, transfer to a sterile fermenter, and cool to room temperature for later use. (3) Under aseptic conditions, add microcapsules of 0.8-2.5% by weight of the extract of *Hydrangea fuciformis* to the fermenter and let it ferment at 35-38°C for 36-50 h. (4) After fermentation, pasteurize in a constant temperature magnetic stirring water bath at 80~85℃ for 10~15 min, then fill and seal.

7. The preparation method according to claim 6, characterized in that: In step (3), the number of live bacteria in the microcapsules is 7 × 10⁻⁶. 7 ~1×10 8 CFU / g.

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