A coconut protein-based fermented cheese containing blueberry seed oil and a method of making the same

Abstract

This invention provides a coconut protein-based fermented cheese containing kumquat seed oil and its preparation method. The steps include: (1) matrix preparation: mixing coconut milk and coconut powder to obtain a mixed matrix; (2) adding kumquat seed oil nanoemulsion to the mixed matrix and stirring evenly; (3) adding lactic acid bacteria starter and CaCl2, fermenting at a constant temperature of 38-42℃, monitoring pH changes, and when the pH drops to 5.5-5.8, adding rennet, continuing to stand at a constant temperature for 40-60 min until curdling is complete, removing whey, and obtaining solids, which is the coconut protein-based fermented cheese containing kumquat seed oil. This invention uses coconut as the sole protein source, completely eliminating the beany taste problem of traditional plant-based cheese. The fat in coconut powder generates characteristic coconut flavor substances through lactone reaction during fermentation and maturation, while the trace terpenes in kumquat seed oil produce a synergistic aroma-enhancing effect with coconut lactones, further enriching the flavor layers of the product.

Description

Technical Field

[0001] This invention relates to the field of food processing technology, specifically to a coconut protein-based fermented cheese containing kumquat seed oil and its preparation method.

[0002] With the rapid growth of the plant-based dairy alternative market, plant-based cheese has gained attention due to its advantages such as meeting the needs of lactose-intolerant individuals and reducing saturated fat and cholesterol intake. Currently, there is research on fermented cheese based on soy, pea, and other legume proteins. However, legume proteins are prone to producing volatile substances such as hexanal and hexenal during fermentation, resulting in a typical beany odor that significantly reduces consumer acceptance.

[0003] Coconut protein, lacking lipoxygenase, does not produce a beany flavor during fermentation. Furthermore, coconut fat is rich in medium-chain fatty acids such as lauric acid, caprylic acid, and capric acid, which can generate characteristic coconut flavor compounds like γ-nonanolactone and δ-decanolactone through lactone formation during fermentation and maturation. However, current coconut fermentation products are mostly concentrated in yogurt or fermented beverages, and research on using coconut as the sole protein source to prepare semi-hard or hard plant-based cheeses remains lacking.

[0004] Kumquat seeds have a high oil content (approximately 35% on a dry basis), with over 73% unsaturated fatty acids in the seed oil. They are also rich in total flavonoids, total limonene, sterols, and other accompanying oil compounds, making them a potential candidate for developing functional oils. However, kumquat seed oil has poor water solubility and low oral bioavailability. Direct addition to fermentation systems can easily lead to oxidation, uneven dispersion, and flavor incompatibility.

[0005] Furthermore, existing technologies lack systematic research on how the composition of solids in coconut substrate affects lactic acid bacteria metabolism, gel network formation, and final product quality by regulating fermentation substrate concentration, protein / fat ratio, and water activity. Summary of the Invention

[0006] Technical problems to be solved: In view of the flavor defects (such as beany taste) and poor stability of functional oils in existing plant-based cheeses, the purpose of this invention is to provide a coconut protein-based fermented cheese containing kumquat seed oil and its preparation method. Using coconut as the sole protein source, by controlling the compound ratio of coconut milk and coconut powder and the stabilizing addition of kumquat seed oil nanoemulsion, the texture and flavor of the product are synergistically optimized, resulting in a semi-hard or hard plant-based cheese with good gel properties, rich coconut flavor and functional nutrients.

[0007] Technical solution: A method for preparing coconut protein-based fermented cheese containing kumquat seed oil, comprising the following steps: (1) Matrix preparation: Mix coconut milk and coconut powder to obtain a mixed matrix; (2) Add the kumquat seed oil nanoemulsion to the mixed matrix and stir evenly; (3) Add lactic acid bacteria starter and CaCl2, ferment at a constant temperature of 38-42℃, monitor the pH value change, when the pH drops to 5.5-5.8, add rennet, continue to stand at a constant temperature for 40-60 min until the curd is complete, remove the whey, and obtain the solids, which is coconut protein-based fermented cheese containing kumquat seed oil.

[0008] Furthermore, the volume-to-mass ratio of coconut milk to coconut powder in step (1) is 1-3 mL: 1 g.

[0009] Furthermore, the volume-to-mass ratio of coconut milk to coconut powder in step (1) is 1 mL: 1 g.

[0010] Furthermore, the preparation method of the kumquat seed oil nanoemulsion in step (2) is any one of the following: System 1 (Tween 80 single system): Using kumquat seed oil as the oil phase, Tween 80 as the emulsifier, and the rest as the aqueous phase, after high-speed shearing and homogenization at 10000-15000 rpm for 2-5 min, the mixture was homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion. System 2 (Tween 80-Sodium Caseinate Composite System): Using kumquat seed oil as the oil phase, sodium caseinate and Tween 80 as emulsifiers, and the remainder as the aqueous phase, the mixture was homogenized by high-speed shearing at 10,000-15,000 rpm for 2-5 min, and then homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion. System 3 (Tween 80-Sodium Caseinate-Mono- and Diglyceride Fatty Acid Ester Ternary Composite System): Using kumquat seed oil as the oil phase, sodium caseinate, Tween 80 and mono- and diglyceride fatty acid esters as emulsifiers, and the remainder as the aqueous phase, after high-speed shearing homogenization at 10000-15000 rpm for 2-5 min, it is homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion (this system has both emulsifying and thickening effects, and is more suitable for high solids fermentation substrates).

[0011] Furthermore, in System 1, the amount of kumquat seed oil is 5 wt.%-10 wt.% of the total emulsion mass, and the amount of Tween 80 added is 0.6 wt.%-3.6 wt.% of the total emulsion mass.

[0012] Furthermore, in System 2, the amount of kumquat seed oil is 5 wt.%-10 wt.% of the total emulsion mass, the amount of sodium caseinate added is 0.5 wt.%-5.0 wt.% of the total emulsion mass, and the amount of Tween 80 added is 1.0 wt.%-2.5 wt.% of the total emulsion mass.

[0013] Furthermore, in the third system, the amount of kumquat seed oil is 5 wt.%-10 wt.% of the total emulsion mass, the amount of sodium caseinate added is 0.5 wt.%-5.0 wt.% of the total emulsion mass, the amount of Tween 80 added is 1.0 wt.%-2.5 wt.% of the total emulsion mass, and the amount of mono- and diglyceride fatty acid esters added is 0.5 wt.%-5.0 wt.% of the total emulsion mass; the mono- and diglyceride fatty acid esters are pre-dissolved in a water bath at 50-70℃ before use.

[0014] Furthermore, the amount of kumquat seed oil nanoemulsion added in step (2) is 1wt.%-8wt.% of the mixed matrix.

[0015] Furthermore, the amount of the kumquat seed oil nanoemulsion added is 5 wt.% of the mixed matrix.

[0016] Furthermore, in step (3), the amount of lactic acid bacteria starter added is 0.25wt.%-1.5wt.%, the amount of CaCl2 added is 0.01wt.%-0.05wt.%, and the amount of rennet added is 0.1wt.%-1.0wt.%.

[0017] The present invention also includes coconut protein-based fermented cheese containing kumquat seed oil prepared by the above method. Beneficial effects

[0018] 1. This invention uses coconut as the sole protein source, completely eliminating the beany taste problem of traditional plant-based cheese. Volatile flavor compound analysis shows that, at a CM:CP ratio of 1:1, the content of coconut aldehyde (γ-nonalactone), a characteristic flavor compound of coconut, reaches 54.12 μg / g, with an odor activity value (OAV) as high as 994.7, which is 2.5 times that of a 5:1 ratio (400.6); the content of δ-decylactone reaches its peak, with an OAV of 31.9, and gamma-octylactone reaches an OAV of 47.6. Creamy aroma lactones account for more than 53%, forming a typical rich coconut and creamy aroma characteristic of this product.

[0019] During the fermentation and maturation process, the fat in coconut powder undergoes lactone formation to generate characteristic coconut flavor compounds. Meanwhile, the trace terpenes in kumquat seed oil have a synergistic aroma-enhancing effect with coconut lactones, further enriching the flavor profile of the product.

[0020] 2. This invention allows for precise adjustment of product hardness within the range of 18-105 g by controlling the blending ratio of coconut milk and coconut powder (Table 1). Specifically, the hardness reaches 104.9 g at a 1:1 ratio, which is 5.6 times that of the 5:1 ratio (18.6 g); the chewiness reaches 29.5, and the gelatinous texture reaches 39.1, both the highest values ​​among all tested ratios. The product at this ratio exhibits typical textural characteristics of semi-hard or hard cheese, with good slicing and spreading properties.

[0021] 3. After nano-encapsulation, the functional components of the kumquat seed oil of this invention are effectively preserved and enhanced in the final product: total flavonoid content ≥15 mg / 100g, total limonene content ≥5 mg / 100g, and unsaturated fatty acids accounting for ≥73% of total fatty acids; simultaneously, the protein content increases significantly with the increase of coconut powder ratio (Table 2), reaching 3.8-3.9 g / 100g in 1:1 and 1:2 ratios, significantly higher than other ratios. Free amino acid analysis (Table 4) shows that with the increase of coconut powder ratio, the content of all 19 free amino acids shows a significant upward trend, with L-arginine content ranking first in all ratios (63.1-221.6 mg / kg), and L-glutamic acid reaching 50.6 mg / kg in the 1:2 ratio, which is 15 times that of the 1:1 ratio (3.3 mg / kg), significantly contributing to the fresh and fragrant flavor of the product.

[0022] 4. The kumquat seed oil of this invention, after being processed by nano-encapsulation technology, exhibits excellent stability during fermentation and storage: (1) Significantly improved oxidative stability: the peroxide value is reduced by more than 50% compared to directly added crude oil; (2) Stable emulsion particle size: no significant change within 30 days of storage at 4℃; (3) Improved flavor stability: the problems of demulsification, oxidation, and flavor conflict caused by direct addition are resolved. In addition, the natural antibacterial activity of limonene can inhibit the growth of miscellaneous bacteria during fermentation, indirectly improving the microbial safety of the product.

[0023] 5. Organic acid analysis in this invention (Table 3) shows that the lactic acid content reaches 3157.4 mg / kg at a 1:1 ratio, the highest among all ratios, exhibiting a typical inverted "U"-shaped trend of first increasing and then decreasing. Lactic acid is the main metabolite of lactic acid bacteria, and its content directly reflects fermentation activity. The moderate content of organic acids such as acetic acid (382.5 mg / kg) and citric acid (1149.9 mg / kg) is beneficial for forming a harmonious flavor profile. Attached Figure Description

[0024] Figure 1 The effect of different Tween addition amounts on the physicochemical properties of kumquat seed oil nanoemulsion in Example 1 is shown; where A is the average particle size; B is the particle size distribution; and C is the polydispersity index (PDI). Note: Different letters represent significant differences between groups (p<0.05). Figure 2 The effect of different Tween addition amounts on the stability of kumquat seed oil nanoemulsion in Example 1; where A is the clarification index and B is the stability coefficient; Figure 3The effect of different sodium caseinate additions on the physicochemical properties of kumquat seed oil nanoemulsion in Example 2 is shown; where A is the average particle size; B is the particle size distribution; and C is the polydispersity index (PDI). Figure 4 The effect of different amounts of sodium caseinate added on the stability of kumquat seed oil nanoemulsion in Example 2; where A is the clarification index and B is the stability coefficient; Figure 5 The effect of different amounts of mono- and diglyceride fatty acid esters added in Example 3 on the physicochemical properties of kumquat seed oil nanoemulsion; where A is the average particle size; B is the particle size distribution; and C is the polydispersity index (PDI). Figure 6 The effect of different amounts of mono- and diglyceride fatty acid esters added on the stability of kumquat seed oil nanoemulsion in Example 3; where A is the clarification index and B is the stability coefficient; Figure 7 The image shows the viscosity and frequency scan of the cheese under different ratios of coconut milk and coconut powder in Example 4; where A represents viscosity and B represents frequency scan. Figure 8 The images show the appearance of the cheese under different ratios of coconut milk and coconut powder in Example 4. Figure 9 The image shows scanning electron microscope (SEM) images of cheese produced in Example 4 with different ratios of coconut milk to coconut powder; Note: magnified 300 times, scale bar is 100 μm. Detailed Implementation

[0025] This invention proposes a coconut protein-based fermented cheese containing kumquat seed oil and its preparation method. To make the objectives, technical solutions, and effects of this invention clearer and more explicit, the following will provide a more detailed description of the invention with reference to specific embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention.

[0026] Lactic acid bacteria starter culture was purchased from Ubit (brand name); it was cultured to a concentration of 1×10⁻⁶ before use. 6 -1×10 7 cfu / mL.

[0027] Example 1 The preparation method of kumquat seed oil nanoemulsion (Tween emulsion stabilized system) includes the following steps: Tween 80 is added as an emulsifier to the emulsion system, with its mass accounting for 0.6%, 1.2%, 1.8%, 2.4%, 3.0%, and 3.6% of the total emulsion mass. 45 mL of distilled water and 5 g of kumquat seed oil are added, and the mixture is homogenized at 10,000 rpm for 3 min, followed by homogenization three times at 115 MPa using a high-pressure microfluidic jet to obtain a stable nanoemulsion.

[0028] Performance testing: 1. Determination of average particle size and potential of nanoemulsions The particle size distribution of the emulsion was determined using laser diffraction. Stagnant, bubble-free distilled water was poured into the sample cell of the laser diffractometer. Spherical particles were selected, and the emulsion was added dropwise to achieve an opacity of 7%-8%. The sample refractive index was 1.414, and the medium refractive index was 1.333. The emulsion was diluted 100-fold with deionized water at pH 7, and its potential was measured using a Zetasizer NanoZSE with an equilibration time of 120 s. All samples were measured at 25°C.

[0029] 2. Determination of the centrifugal stability and clarification index of nanoemulsions Emulsions with different CMPI concentrations were placed in LUM sample tubes and centrifugal stability and clarification index were determined using a LUMiSizer stability analyzer. The wavelength was set to 870 nm and the optical factor was 1.00. The contour lines were set to 500. The samples were centrifuged at 3000 rpm for 50 min, and the transmittance curves were recorded every 5 s.

[0030] The results are as follows Figure 1 As shown in Figure A, as the concentration of Tween-80 increased from 0.6% to 3.6%, the average particle size of the emulsion decreased from 215.5 nm to approximately 155.4 nm. Tween, as a nonionic emulsifier, significantly reduced the interfacial tension of the system, forming a dense protective film on the droplet surface. This film effectively suppressed droplet collisions and aggregation through a strong steric hindrance effect, thereby maintaining the microscopic stability of the system. Figure 1 The particle size distribution curve of group B further corroborates the above mechanism. The distribution peak of the low concentration group (0.6%) is broader and shows signs of right shift, reflecting that a small amount of emulsifier is insufficient to completely cover the newly formed interface, causing some droplets to aggregate under Brownian motion and exhibit polydisperse characteristics.

[0031] like Figure 2 A represents the clarifying index of the emulsion. When the Tween addition amount was 0.6%, the clarifying index was the lowest (0.007), significantly lower than other treatment groups. P<0.05). As the concentration increases to 1.8%-2.4%, the index decreases significantly, reaching its lowest value (0.022) in the 2.4% group, exhibiting optimal stability. The stability coefficient R value measured by the colorimetric-turbidity method is consistent with the above trend: the R value increases significantly from 0.45 with increasing concentration, reaching a peak value (0.99) at 2.4%, and then decreases slightly in the high concentration range. This is because under low concentration (0.6%) conditions, the sparse surfactant molecules are insufficient to completely cover the interface of newly formed oil droplets, resulting in high interfacial tension, making oil droplets prone to collision, aggregation, and migration. With increasing concentration, Tween molecules form a highly dense and ordered adsorption layer at the interface, effectively blocking the contact between oil droplets through the strong steric repulsion force generated by the polyoxyethylene chains.

[0032] Example 2 The preparation method of kumquat seed oil nanoemulsion (Tween-sodium caseinate emulsion stabilization system) includes the following steps: sodium caseinate is added to beakers at 0.5%, 1.0%, 2.0%, 3.0%, 4.0%, and 5.0% of the total emulsion mass, respectively. Distilled water is added to make the total mass of sodium caseinate and distilled water in each group 43.8 g. The mixture is magnetically stirred for 2 h to fully dissolve the sodium caseinate in the distilled water. The pH is adjusted to 7.0. Then, 1.2 g of Tween and 5 g of kumquat seed oil are added. After homogenization at 10,000 rpm for 3 min, the mixture is homogenized three times with a high-pressure microfluidic jet at 115 MPa to obtain a stable nanoemulsion.

[0033] The results are as follows Figure 3 As shown in Figure A, with the addition of sodium caseinate increasing from 0.5% to 3.0%, the average particle size of the emulsion showed a significant decreasing trend, decreasing significantly from the initial 110.2 nm to 90.8 nm. P <0.05). For example... Figure 3 As shown in B, all treatment groups maintained a unimodal distribution, but the peak shape of the 2.0%-3.0% group was the narrowest and sharpest, exhibiting high homogeneity; the distribution curve of the high concentration group (5.0%) showed a significantly broadened base. Figure 3 As shown in C, the PDI of the 2.0% group reaches a minimum (approximately 0.14), while when the concentration exceeds 3.0%, the PDI rises significantly to above 0.18, indicating that the particle dispersion uniformity of the system has decreased.

[0034] The effect of different sodium caseinate additions on the kinetic stability of nanoemulsions is as follows: Figure 4 As shown in A, when the sodium caseinate concentration was in the range of 0.5%-2.0%, the clarification index was extremely low and remained stable (0.02), with no significant difference between groups. P>0.05); However, as the concentration further increased to above 3.0%, the clarification index increased significantly, with the 5.0% group jumping to a peak value (0.37), indicating that the system's resistance to centrifugal stress weakened. Figure 4 The stability coefficient R of B corroborates the above trend: in the concentration range of 1.0%-3.0%, the R value remains consistently above 0.8, reaching its maximum at 3.0%; subsequently, it decreases rapidly at 4.0% and 5.0%, reflecting that emulsion particles are more prone to sedimentation or flocculation under strong centrifugal fields in high protein concentration environments. The underlying mechanism of this initial stability followed by a decrease in stability stems from the physical state of proteins in the aqueous phase. In the low concentration range (<2.0%), sodium caseinate mainly exists as monomers or micromicelles and is adsorbed at the oil-water interface, maintaining the static equilibrium of the system through high absolute value charge repulsion. When the concentration exceeds the saturation adsorption threshold of 3.0%, excess protein molecules in the continuous phase begin to self-assemble into larger aggregates through hydrophobic interactions.

[0035] Example 3 The preparation method of kumquat seed oil nanoemulsion (Tween-sodium caseinate-mono- and diglyceride fatty acid ester emulsion stabilized system) includes the following steps: Monoglyceride and diglyceride fatty acid esters were added to a beaker at 0.5%, 1.0%, 2.0%, 3.0%, 4.0%, and 5.0% of the total emulsion mass, respectively. Then, 1% sodium caseinate was added, and distilled water was added to make the total mass 43.8 g. The mixture was magnetically stirred in a water bath at 60°C for 2 h to fully dissolve the emulsifier in the distilled water, and the pH was adjusted to 7.0. 1.2 g of Tween and 5 g of calamari seed oil were then added. After homogenization at 10,000 rpm for 3 min, the mixture was homogenized three times with a high-pressure microfluidic jet at 115 MPa to obtain a stable nanoemulsion.

[0036] like Figure 5 As shown, different amounts of mono- and diglyceride fatty acid esters significantly affected the emulsion particle size, PDI, and particle size distribution. With the addition amount increasing from 0.5% to 5.0%, the average particle size of the emulsion generally showed a trend of first a small change followed by a significant increase. The particle size was relatively small at the 1.0% addition amount, while it significantly increased at 4.0% and 5.0%. The PDI results indicated some differences in dispersibility among the treatment groups. The highest PDI value was observed at the 2.0% addition amount, indicating a wider system distribution and poorer uniformity. The 1.0% and 3.0% treatment groups showed relatively lower PDIs, indicating a more homogeneous and stable emulsion system.

[0037] like Figure 6As shown in Figure A, as the addition amount increased from 0.5% to 5.0%, the emulsion clarification index generally showed a trend of first decreasing and then increasing, reaching its lowest value at an addition amount of 2.0%, indicating that the system was most stable at this point. When the addition amount continued to increase to 4.0% and 5.0%, the clarification index increased significantly, especially in the 5.0% group, indicating that the emulsion underwent more obvious aggregation or phase separation. Figure 6 As shown in Figure B, the trend of R value changes corresponds to the clarification index: the R value is higher at addition levels of 1.0% and 2.0%, indicating that the emulsion structure is relatively stable.

[0038] The above examples explored the effects of nonionic surfactants (Tween), amphiphilic proteins (sodium caseinate), and lipophilic emulsifiers (monoglycerides and diglycerides) on the physicochemical properties of nanoemulsions, revealing the stabilization mechanisms of different systems and establishing optimal formulations: Tween mainly relies on the hydrated steric hindrance layer formed by polyoxyethylene chains to maintain stability, achieving optimal kinetic stability at an addition of 2.4%, exhibiting significant viscous fluid characteristics; sodium caseinate utilizes the electrostatic repulsion force generated by high-density surface negative charges to construct microdroplets, with an optimal concentration range of 1%, while excessive addition (>3.0%) induces instability and gel-like structural transformation due to continuous phase protein aggregation; while monoglycerides and diglycerides provide physical support by constructing a microcrystalline network in the continuous phase, endowing the emulsion with a strong elastic response, with an optimal addition of 2.0%, while excessive addition easily induces excessive crystal growth and aggregation. In summary, this study clarified the differences in the contributions of various emulsifiers in terms of steric hindrance, electrostatic repulsion, and structured continuous phase, laying a crucial formulation foundation for the subsequent construction of high-performance compound nanoemulsion systems.

[0039] Example 4: Coconut milk and coconut powder in different volume-to-mass ratios A method for preparing coconut protein-based fermented cheese containing calamansi seed oil includes the following steps: (1) Matrix preparation: Coconut milk (CM) and coconut powder (CP) are mixed in volume-to-mass ratios of 5mL:1g, 3mL:1g, 2mL:1g, 1mL:1g, and 1mL:2g respectively (total mass of the mixture is 120g). The mixture is magnetically stirred for 25min to ensure uniform mixing and obtain the mixed matrix. (2) Add 6g of kumquat seed oil nanoemulsion (selecting the best conditions in Example 3, i.e., Tween 2.4%, sodium caseinate 1%, mono- and diglycerides 2.0%) to the mixed matrix and stir evenly; (3) Add 1.0 wt.% lactic acid bacteria starter and 0.02 wt.% CaCl2, and ferment at a constant temperature of 40℃. Measure the pH value every hour. When the pH drops to between 5.5 and 5.8, add 0.5 wt.% rennet and stir magnetically for 15 minutes to dissolve the rennet. Continue to stand at a constant temperature for 50 minutes until the curdling is complete. Remove the whey and obtain the solids, which are coconut protein-based fermented cheese containing kumquat seed oil.

[0040] Performance testing: 1. Solid yield The solids yield (%) is obtained by dividing the solids content after filtering the whey by the total mass. The calculation formula is as follows: ; Where m is the mass of the solid (g), m 总 The total mass of the sample is (g).

[0041] 2. Rheological properties The effects of dynamic rheometer on the apparent viscosity, viscoelastic modulus, thixotropy, and creep recovery properties of samples before whey removal after fermentation were investigated. The effects were studied within the range of 0.1–100 s⁻¹. -1 Apparent viscosity was tested within the specified range. Elastic modulus and viscous modulus were measured in frequency sweeps within the range of 0.1–10 Hz (strain 1%). The creep recovery properties of the gel were characterized using a three-segment thixotropic test: samples were scanned over time at alternating strain amplitudes (1% and 100%) for durations of 60 s, 80 s, and 100 s, respectively. After the thixotropic test, a fixed stress of 3 Pa (within the linear range) was applied to the sample for 300 s, then the applied stress was removed, and the creep recovery properties were recorded for another 600 s.

[0042] 3. Texture determination The texture properties of the solids after whey removal, such as hardness, chewiness, and cohesiveness, were analyzed using the TPA mode of a texture analyzer. The parameters of the texture analyzer were set as follows: probe descent speed before testing was 4.0 mm / s, testing speed was 1.0 mm / s, probe return speed after testing was 1.0 mm / s, downward deformation was 30%, trigger force was 10 g, and probe type was P / 35.

[0043] 4. Scanning electron microscope After freeze-drying the solid, the microstructure of the sample was observed using scanning electron microscopy (SEM). The SEM results were analyzed using ImageJ 1.52p (Bharti Airtel Ltd, India) software to calculate the porosity of the gel sample. Porosity is expressed as the ratio between the pore area and the total area of ​​the image.

[0044] 5. Determination of basic physicochemical indicators (1) Protein content was determined by the Kjeldahl method (GB / T 5009.5-2016), and each treatment group was measured in parallel three times.

[0045] (2) The pH value was measured by mixing 10 mL of anhydrous CO2 with 1 g of ground cheese thoroughly and then using a pH meter. Each treatment group was measured in parallel 3 times.

[0046] (3) The fat content in cheese was determined by Soxhlet extraction according to the national standard (GB5009.6-2016), and each treatment group was measured in parallel 3 times.

[0047] (4) Moisture content was determined by direct drying method (GB 5009.3-2016), with each treatment group measured in parallel 3 times.

[0048] 6. Determination of organic acids Accurately weigh 1.2 g of sample into a 10 mL centrifuge tube, then add 8.0 mL of 0.01 mol / L H₂SO₄ solution. Vortex the mixture for 60 s and centrifuge at 3500 rpm for 15 min. Filter the supernatant using a 0.22 μm filter and store at 4 °C for later use. The organic acid determination parameters are as follows: column: Spursil C18 column; mobile phase: methanol and potassium dihydrogen phosphate buffer (0.1 mol / L, pH 2.7) in a ratio of 3:97 (v / v); UV detection wavelength: 210 nm; flow rate: 1.0 mL / min; column temperature: 35 °C; injection volume: 20 μL.

[0049] 7. Determination of free amino acids Determination of free amino acids: Take 5g of cheese, add 50mL of water, homogenize, centrifuge, take 5mL of supernatant, add xanthanyl salicylic acid (final concentration of 7%), let stand overnight at 4℃, centrifuge, take the supernatant and freeze dry for 48h, seal the obtained dried powder, and determine according to GB / T5009.124-2016.

[0050] 8. Determination of volatile flavor compounds Headspace solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC / MS) was used to determine the types and contents of volatile substances in the sample. 2.0 g of sample was placed in a 15 mL extraction flask, and 2.0 µL of 0.21 µg / µL 2-octanol was added as an internal standard (IS). Extraction was performed at 55 °C for 30 min using a 65 μm PDMS / DVB fiber. After extraction, the fiber was removed and inserted into the gas chromatography vaporization chamber for GC-MS analysis. Samples were analyzed three times for each maturation time.

[0051] GC conditions: A DB-WAX capillary column (30m × 0.25mm, 0.25μm) was used, with He as the carrier gas and a flow rate of 1mL / min. The column temperature program was as follows: initial column temperature 35℃, hold for 4min, increase to 100℃ at 4℃ / min, hold for 2min, then increase to 150℃ at 3℃ / min, and finally increase to 230℃ at 10℃ / min, hold for 2min. The injection port temperature was 250℃, and splitless injection was used.

[0052] MS conditions: electron ionization source, ionization energy 70 eV, ion source temperature 230℃, interface temperature 250℃, quadrupole temperature 150℃, detector temperature 280℃, scan mode full scan, mass range 20-350 amu.

[0053] GC-MS data analysis: Qualitative analysis of volatile components in the samples was performed using a computer spectral library (NIST11 / Wiley7.0), and the retention indices of each chromatographic peak were calculated using the retention times of C8–C20 n-alkanes to confirm the chemical composition of the volatile substances. Quantitative analysis of volatile components was performed using the area normalization method.

[0054] Cluster analysis and principal component analysis were used to identify differences in volatile components among different cheeses. Cluster analysis was performed using R software, while principal component analysis (PCA) and correlation analysis between cheese characteristic aroma compounds and sensory scores (PLSR analysis) were performed using Unscrambler 9.7.

[0055] Test results: 1. Rheological properties Depend on Figure 7 As shown in Figure A, the apparent viscosity of all samples decreased significantly with increasing shear rate, exhibiting typical shear-thinning behavior. This indicates that the cheese system possesses pseudoplastic fluid characteristics, which is related to its protein- and fat-rich emulsion-filled gel network structure. The same shear-thinning phenomenon was also observed in coconut milk-based fermented yogurt with added tapioca starch or soy protein isolate*.

[0056] In the low shear rate region (γ̇ < 30 s⁻¹), the viscosity values of cheese samples gradually decreased with the increase in the proportion of coconut powder. Combining with the previous physicochemical data (Table 1), this result indicates that the CM:CP 2:1 formula may form a relatively weak flocculation network that deconstructs even at extremely low shear rates. In contrast, although the viscosity of cheese with CM:CP 1:1 is lower than that of CM:CP 5:1 and 3:1, its viscosity exceeds other samples in the high shear rate region (γ̇ > 40 s⁻¹). This result reveals that the CM:CP 1:1 formula has a stronger shear thickening or structure reconstruction ability, which may be related to the dense emulsion network formed by its high fat content (36.0%) and moderate moisture content (37.2%).

[0057] Figure 7 Figure B shows that G′ and G″ of all proportion samples increase with the increase in frequency, and G′ is always greater than G″ throughout the frequency range (except in the extremely low frequency region), indicating that the cheese forms a weak gel network dominated by elasticity. The fermented coconut milk-based yogurt added with tapioca starch or soy protein isolate also shows a viscoelastic gel structure (G′ > G″)*.

[0058] However, in the lowest frequency (0.16 Hz) region, the G″ of cheese samples with CM:CP 5:1, 3:1, and 2:1 is slightly higher than G′ (tanδ > 1), showing a viscous response, while the G′ > G″ (tanδ < 1) of cheese samples with CM:CP 1:1 and 1:2, indicating that the latter two have a more stable gel structure. The differences in rheological behaviors can be attributed to the synergistic effects of the interactions between protein-fat networks caused by fermentation at different CM:CP ratios. At low coconut powder ratios (5:1, 3:1), the relatively weak gel network formed by the lower protein concentration is rapidly destroyed under high-speed shear. Therefore, in the low frequency region, the dynamic modulus shows G′ < G″, confirming that its gel network structure is mainly viscous. With the increase in the proportion of coconut powder, fermentation promotes sufficient cross-linking between proteins in the cheese, and fat globules are distributed as fillers, which is conducive to the formation of a continuous gel network. Therefore, it shows G′ > G″, indicating the formation of a quasi-solid (gel)*. This also explains why the cheese sample with CM:CP 1:1 has the highest yield and the best texture. However, when the coconut powder ratio is too high, it restricts the fermentation of the starter culture, and there is a lack of slow cross-linking of proteins induced by acid in the cheese, making the network mainly composed of physical entanglements rather than chemical bonding. Therefore, the rheological behavior of the cheese sample with CM:CP 1:2 shows a medium modulus, but the viscosity is unstable under high shear.

[0059] 2. Solids yield and texture properties Table 1 Texture of cheese at different ratios of coconut milk to coconut powder CM:CP hardness Adhesion elasticity Cohesion gelatinous chewing toughness 5:1 202.599 -28.001 0.760 0.338 6.296 4.993 0.044 3:1 132.939 -45.678 0.722 0.366 13.968 10.218 0.024 2:1 311.856 -32.155 0.818 0.509 13.955 12.626 0.049 1:1 818.007 -91.534 0.727 0.389 39.054 29.464 0.037 1:2 227.177 -42.000 0.827 0.435 15.476 13.564 0.034 As shown in Table 1 above, when the CM:CP ratio is 1:1, the product's hardness, gelatinous texture, chewiness, and adhesiveness all reach their maximum values, forming a firm gel texture. This ratio is suitable for developing semi-hard or hard plant-based cheeses, characterized by good slicing properties and a strong chewy texture.

[0060] When the CM:CP ratio is 1:2, hardness, gelatinous texture, and chewiness all decrease significantly, indicating that the low moisture environment (<30%) caused by excessive coconut powder disrupts the continuity of the gel network, making the product brittle rather than tough. This ratio is not suitable for products that require a dense texture, but it can be considered for the development of spreadable or soft products.

[0061] 3. Appearance and morphology and scanning electron microscopy Depend on Figure 8 As can be seen, the cheese is generally milky white or milky yellow, forming a texture similar to semi-hard cheese, and has a certain degree of plasticity. Figure 9 It is known that cheese exhibits an emulsion-filled gel structure, with fat particles encapsulated within a three-dimensional gel network formed by proteins or proteoglycans, creating a continuous structure. This is similar to the microstructure of other plant-based or mixed-protein cheeses. As the proportion of coconut flour in the cheese increases, the gel network becomes more tightly connected, which may explain the increased cheese yield. Coconut flour contains not only coconut protein but also a large amount of fiber, which further facilitates the formation of a gel network to encapsulate fat particles.

[0062] 4. Basic physicochemical indicators Table 2 Physicochemical properties of cheese with different coconut milk to coconut powder (CM:CP) ratios CM:CP 5:1 3:1 2:1 1:1 1:2 Protein content (g / 100g) <![CDATA[3.0±0.0 c ]]> <![CDATA[3.2±0.1 bc ]]> <![CDATA[3.4±0.0 b ]]> <![CDATA[3.8±0.2 a ]]> <![CDATA[3.9±0.2 a ]]> Fat content (g / 100g) <![CDATA[20.8±1.3 d ]]> <![CDATA[27.0±1.0 c ]]> <![CDATA[25.2±1.0 c ]]> <![CDATA[36.0±1.8 a ]]> <![CDATA[30.5±0.4 b ]]> Moisture content (%) <![CDATA[56.8±1.4 a ]]> <![CDATA[54.8±1.3 a ]]> <![CDATA[50.0±0.4 b ]]> <![CDATA[37.2±1.4 c ]]> <![CDATA[29.9±0.9 d ]]> Yield (%) <![CDATA[13.7±4.2 e ]]> <![CDATA[23.2±5.8 d ]]> <![CDATA[26.9±5.3 c ]]> <![CDATA[41.2±1.7 a ]]> <![CDATA[30.2±1.4 b ]]> Note: Different lowercase letters indicate that different ratios of coconut milk to coconut powder are significant. p<0.05 ) As shown in Table 2, the protein content in the product increased significantly with the increase in the proportion of coconut powder (CM:CP decreased from 5:1 to 1:2) (p<0.05). The protein content reached its highest level (3.8–3.9 g / 100g) when the CM:CP ratio was 1:1 and 1:2, significantly higher than the 5:1, 3:1, and 2:1 sample groups. This indicates that increasing the amount of coconut powder can effectively improve the protein nutritional density of the final product.

[0063] The fat content initially increased and then slightly decreased with increasing coconut powder ratio. At a CM:CP ratio of 1:1, the fat content peaked (36.0 g / 100g), significantly higher than all other ratios (p<0.05). When the ratio further decreased to 1:2, the fat content dropped to 30.5 g / 100g, but remained significantly higher than the 5:1-2:1 groups. This change indicates that there is an optimal range for coconut powder's contribution to fat content; excessive addition may lead to a decrease in relative fat content.

[0064] The moisture content showed a continuous and significant decreasing trend with increasing coconut powder ratio (p<0.05). The moisture content was highest (56.8%) when the CM:CP ratio was 5:1, and lowest (29.9%) when the ratio was 1:2. The decrease in moisture content was directly related to the increase in total solids content in the system after coconut powder replaced coconut milk, indicating that adjusting the coconut milk / coconut powder ratio can effectively control the water activity and texture of the product.

[0065] 5. Organic acids Table 3. Organic acid content of cheese at different ratios of coconut milk and coconut powder CM:CP 5:1 3:1 2:1 1:1 1:2 oxalic acid 13.±1.8 <![CDATA[16.4±1.4 bc ]]> <![CDATA[16.6±0.7 bc ]]> <![CDATA[19.8±0.4 b ]]> <![CDATA[49.3±4.3 a ]]> tartaric acid <![CDATA[39.0±2.0 a ]]> <![CDATA[62.7±3.0 b ]]> <![CDATA[65.1±1.36 b ]]> <![CDATA[62.4±3.9 b ]]> <![CDATA[90.4±6.5 a ]]> malic acid <![CDATA[1859.5±40.1 e ]]> <![CDATA[2562.6±30.5 d ]]> <![CDATA[2955.8±81.6 b ]]> <![CDATA[2780.6±26.7 c ]]> <![CDATA[9796.6±111.3 a ]]> lactic acid <![CDATA[2773.6±66.8 c ]]> <![CDATA[2955.5±53.2 b ]]> <![CDATA[3113.2±17.5 a ]]> <![CDATA[3157.4±29.7 a ]]> <![CDATA[1684.4±28.3 d ]]> Acetic acid <![CDATA[140.0±18.1 d ]]> <![CDATA[186.0±25.4 d ]]> <![CDATA[474.6±14.9 b ]]> <![CDATA[382.5±15.4 c ]]> <![CDATA[617.9±782.9 a ]]> Citric acid <![CDATA[680.7±19.7 e ]]> <![CDATA[880.7±17.9 d ]]> <![CDATA[1026.5±70.2 c ]]> <![CDATA[1149.9±14.8 b ]]> <![CDATA[1321.2±58.8 a ]]> Succinic acid <![CDATA[169.8±4.4 a ]]> <![CDATA[175.5±19.7 a ]]> <![CDATA[115.6±9.1 b ]]> <![CDATA[131.0±13.6 b ]]> <![CDATA[166.6±11.5 a ]]> Note: Different lowercase letters indicate that different ratios of coconut milk to coconut powder are significant. p<0.05 ) As shown in Table 3, with the increase in the proportion of coconut powder (CM:CP decreasing from 5:1 to 1:2), the contents of malic acid, citric acid, tartaric acid, and oxalic acid all showed a continuous and significant upward trend (p<0.05). Malic acid showed the most significant increase, rising from 1859.5 mg / kg (5:1) to 9796.6 mg / kg (1:2), an increase of approximately 5.3 times. Citric acid increased from 680.7 mg / kg to 1321.2 mg / kg, an increase of approximately 1.9 times. Oxalic acid at the 1:2 ratio climbed sharply to 49.3 mg / kg, significantly higher than other groups. Tartaric acid reached a peak of 90.4 mg / kg at the 1:2 ratio.

[0066] Lactic acid content showed an inverted "U"-shaped trend, first increasing and then decreasing significantly. The lactic acid content was highest (3157.4 mg / kg) when the CM:CP ratio was 1:1, but when the ratio was further increased to 1:2, the lactic acid content plummeted to 1684.4 mg / kg, significantly lower than the 5:1 control group (p<0.05).

[0067] The acetic acid content generally increased with the increase of the proportion of coconut powder, but it dropped in stages at the 1:1 ratio, and then reached the highest value of 617.9 mg / kg at the 1:2 ratio.

[0068] Succinic acid content decreased significantly at CM:CP ratios of 2:1 and 1:1 (115.6–131.0 mg / kg), while it remained at higher levels at ratios of 5:1, 3:1, and 1:2 (166.6–175.5 mg / kg). This variation suggests that moderate amounts of coconut flour may inhibit the succinic acid formation pathway, while extreme ratios may promote its accumulation.

[0069] 6. Free amino acids Table 4. Free amino acid content of cheese at different ratios of coconut milk and coconut powder CM:CP 5:1 3:1 2:1 1:1 1:2 L-Aspartic acid na na <![CDATA[28.1±1.3 c ]]> <![CDATA[30.4±1.0 b ]]> <![CDATA[41.3±0.6 a ]]> L-glutamic acid na na na <![CDATA[3.3±0.2 b ]]> <![CDATA[50.6±1.2 a ]]> L-serine na na <![CDATA[1.0±0.1 b ]]> <![CDATA[5.5±0.2 a ]]> <![CDATA[5.7±0.03 a ]]> glycine <![CDATA[7.02±0.2 e ]]> <![CDATA[8.05±0.4 d ]]> <![CDATA[11.3±0.4 c ]]> <![CDATA[18.7±0.5 b ]]> <![CDATA[34.9±0.6 a ]]> L-histidine <![CDATA[4.2±0.1 e ]]> <![CDATA[5.7±0.04 d ]]> <![CDATA[12.4±0.2 c ]]> <![CDATA[20.06±0.6 b ]]> <![CDATA[61.7±0.6 a ]]> L-arginine <![CDATA[63.1±1.4 e ]]> <![CDATA[86.5±1.2 d ]]> <![CDATA[139.7±0.9 c ]]> <![CDATA[155.03±5.03 b ]]> <![CDATA[221.6±1.5 a ]]> L-threonine <![CDATA[0.7±0.1 d ]]> <![CDATA[1.5±0.1 d ]]> <![CDATA[5.2±0.2 c ]]> <![CDATA[9.52±0.74 b ]]> <![CDATA[19.1±0.5 a ]]> L-alanine <![CDATA[21.7±0.5 d ]]> <![CDATA[23.03±0.2 d ]]> <![CDATA[33.3±0.4 c ]]> <![CDATA[42.6±1.7 b ]]> <![CDATA[77.8±0.8 a ]]> L-proline <![CDATA[22.0±1.3 e ]]> <![CDATA[26.0±0.3 d ]]> <![CDATA[33.8±0.6 c ]]> <![CDATA[38.01±0.9 b ]]> <![CDATA[52.3±1.0 a ]]> L-tyrosine <![CDATA[30.3±0.6 e ]]> <![CDATA[39.4±0.1 d ]]> <![CDATA[46.0±0.7 c ]]> <![CDATA[50.9±1.1 b ]]> <![CDATA[59.7±1.0 a ]]> L-valine na <![CDATA[1.2±0.1 d ]]> <![CDATA[4.5±0.2 c ]]> <![CDATA[10.8±0.4 b ]]> <![CDATA[46.6±0.2 a ]]> L-methionine <![CDATA[2.2±0.1 e ]]> <![CDATA[6.2±0.1 d ]]> <![CDATA[3.07±0.09 c ]]> <![CDATA[5.09±0.06 b ]]> <![CDATA[18.7±0.2 a ]]> L-cysteine <![CDATA[56.7±0.5 c ]]> <![CDATA[57.8±0.1 b ]]> <![CDATA[57.7±0.1 b ]]> <![CDATA[57.03±0.7 bc ]]> <![CDATA[59.2±0.2 a ]]> L-Isoleucine <![CDATA[1.1±0.07 d ]]> <![CDATA[2.7±0.1 c ]]> <![CDATA[2.3±0.03 c ]]> <![CDATA[4.7±0.1 b ]]> <![CDATA[33.6±0.6 a ]]> L-Leucine <![CDATA[5.2±0.3 e ]]> <![CDATA[9.9±0.3 d ]]> <![CDATA[13.9±0.3 c ]]> <![CDATA[18.0±0.5 b ]]> <![CDATA[77.1±1.0 a ]]> L-phenylalanine <![CDATA[10.0±0.3 e ]]> <![CDATA[13.4±0.3 d ]]> <![CDATA[17.41±0.3 c ]]> <![CDATA[20.1±0.6 b ]]> <![CDATA[104.9±0.4 a ]]> L-Tryptophan <![CDATA[6.2±0.1 e ]]> <![CDATA[10.4±0.1 c ]]> <![CDATA[9.2±0.1 d ]]> <![CDATA[11.6±0.01 b ]]> <![CDATA[29.2±0.3 a ]]> L-Lysine <![CDATA[1.3±0.08 e ]]> <![CDATA[2.5±0.1 d ]]> <![CDATA[9.6±0.1 c ]]> <![CDATA[16.4±0.4 b ]]> <![CDATA[54.4±0.5 a ]]> Note: Different lowercase letters indicate that different ratios of coconut milk to coconut powder are significant. p<0.05 ) As shown in Table 4, with the increase of the proportion of coconut powder (CM:CP decreased from 5:1 to 1:2), the contents of all 19 free amino acids detected showed a continuous and significant upward trend (p<0.05), reaching the highest value at the 1:2 ratio. This trend is consistent with the change in protein content in Table 1, indicating that coconut powder is the main source or precursor contributor of free amino acids.

[0070] At 5:1 and 3:1 ratios, L-aspartic acid, L-glutamic acid, L-serine, and L-valine were not detected (na). These amino acids only became detectable when the CM:CP ratio reached 2:1 or 1:1, and peaked at 1:2. This indicates that there is a minimum threshold (approximately 2:1) for the amount of coconut powder added; below this threshold, some characteristic amino acids cannot be effectively released or accumulate.

[0071] L-Glutamic acid: The content jumps to 50.6 mg / kg in a 1:2 ratio, which is 15 times that in a 1:1 ratio. As a major contributor to umami flavor, its significant increase is crucial to enhancing the umami flavor of the product.

[0072] L-Phenylalanine, L-Leucine, and L-Valine: These are branched-chain and aromatic amino acids, reaching 104.9, 77.1, and 46.6 mg / kg respectively in a 1:2 ratio, showing a significant increase. These amino acids are precursors for subsequent flavor transformations (transamination and decarboxylation) and are crucial for the formation of malty, floral, and fruity aromas during cheese maturation.

[0073] L-arginine: It has the highest content in all proportions (63.1-221.6 mg / kg) and is the most abundant free amino acid in coconut plant-based cheese, which may be related to the inherent composition of coconut raw materials.

[0074] 7. Volatile flavor compounds Table 5. Relative content of volatile substances in cheese at different ratios of coconut milk to coconut powder.

[0075] Table 6. Volatile substance content of cheese at different ratios of coconut milk to coconut powder

[0076] Note: OVA values ​​<1 can be ignored, meaning they have no "contribution" to flavor.

[0077] As shown in Table 5-7, the ratio of coconut milk to coconut powder is a key variable controlling the flavor type of plant-based cheese: Low coconut powder (CM:CP of 5:1-3:1): esters, aldehydes, and terpenes are relatively high, with a flavor leaning towards fruity, green, and citrus aromas, and an overall light fragrance. Medium coconut powder (CM:CP of 2:1-1:1): lactones are absolutely dominant (>53%), with coconut aldehyde, δ-decyl lactone, and OAV reaching their peak levels. The flavor shifts towards a rich coconut and creamy aroma, representing the most prominent range of coconut characteristic flavors. High coconut powder (CM:CP of 1:2): acid content surges to 69%, lactones decline, and the flavor exhibits a strong acidity and fattiness, accompanied by a resurgence of citrus / grassy aromas from octanal.

[0078] At a CM:CP ratio of 1:1, the coconut aldehyde OAV (994.7 g / g) and absolute content (54.12 μg / g) were both the highest, and key lactones such as δ-decanolide and propyloctanolide also reached their peak values. Combining the results in Tables 1-4 showing the strongest texture and highest lactic acid fermentation activity at this ratio, the 1:1 ratio is the optimal balance point for achieving the triple goals of "rich coconut aroma + dense texture + fermentation activity".

[0079] Based on the screening criterion of OAV>1, 11 compounds, including coconut aldehyde, δ-decanoic acid lactone, gamma-octanoic acid lactone, ethyl decanoate, ethyl octanoate, nonanal, octanal, and doxy-dodecanoic acid lactone, were identified as key flavor active substances in this product. Among them, coconut aldehyde contributes more than 80% of OAV and is the primary characteristic aroma component of the product.

[0080] 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 method for preparing a coconut protein-based fermented cheese containing aronia seed oil, characterized by, Includes the following steps: (1) Matrix preparation: Mix coconut milk and coconut powder to obtain a mixed matrix; (2) Add the kumquat seed oil nanoemulsion to the mixed matrix and stir evenly; (3) Add lactic acid bacteria starter and CaCl2, ferment at a constant temperature of 38-42℃, monitor the pH value change, when the pH drops to 5.5-5.8, add rennet, continue to stand at a constant temperature for 40-60 min until the curd is complete, remove the whey, and obtain the solids, which is coconut protein-based fermented cheese containing kumquat seed oil.

2. A method of preparing a coconut protein based fermented cheese containing Vaccinium vitis-idaea seed oil according to claim 1, characterized in that, The volume-to-mass ratio of coconut milk to coconut powder in step (1) is 1-3 mL: 1 g.

3. A method of preparing a coconut protein based fermented cheese containing phyllium seed oil according to claim 1, characterized in that, The volume-to-mass ratio of coconut milk to coconut powder in step (1) is 1 mL: 1 g.

4. A method of preparing a coconut protein based fermented cheese containing phyllium seed oil according to claim 1, characterized in that, The preparation method of the kumquat seed oil nanoemulsion in step (2) is any one of the following: System 1: Using kumquat seed oil as the oil phase, Tween 80 as the emulsifier, and the remainder as the aqueous phase, the mixture was homogenized by high-speed shearing at 10,000-15,000 rpm for 2-5 min, and then homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion. System 2: Using kumquat seed oil as the oil phase, sodium caseinate and Tween 80 as emulsifiers, and the remainder as the aqueous phase, the mixture was homogenized by high-speed shearing at 10,000-15,000 rpm for 2-5 min, and then homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion. System 3: Using kumquat seed oil as the oil phase, sodium caseinate, Tween 80 and mono- and diglycerides of fatty acids as emulsifiers, and the remainder as the aqueous phase, the mixture is homogenized by high-speed shearing at 10,000-15,000 rpm for 2-5 min, and then homogenized 2-4 times at 100-120 MPa using high-pressure microfluidic jet to obtain a stable nanoemulsion.

5. A method of preparing a coconut protein based fermented cheese containing Vaccinium uliginosum seed oil according to claim 4, characterized in that, In System 1, the amount of kumquat seed oil is 5 wt.%-10 wt.% of the total emulsion mass, and the amount of Tween 80 added is 0.6 wt.%-3.6 wt.% of the total emulsion mass.

6. A method of preparing a coconut protein based fermented cheese containing phyllium seed oil according to claim 4, characterized in that, In System 2, the amount of kumquat seed oil is 5 wt.%-10 wt.% of the total emulsion mass, the amount of sodium caseinate added is 0.5 wt.%-5.0 wt.% of the total emulsion mass, and the amount of Tween 80 added is 1.0 wt.%-2.5 wt.% of the total emulsion mass.

7. A method of preparing a coconut protein based fermented cheese containing phyllium seed oil according to claim 4, characterized in that, In the system described above, the amount of kumquat seed oil in the system is 5 wt.%-10 wt.% of the total emulsion mass, the amount of sodium caseinate added is 0.5 wt.%-5.0 wt.% of the total emulsion mass, the amount of Tween 80 added is 1.0 wt.%-2.5 wt.% of the total emulsion mass, and the amount of mono- and diglyceride fatty acid esters added is 0.5 wt.%-5.0 wt.% of the total emulsion mass; the mono- and diglyceride fatty acid esters are pre-dissolved in a water bath at 50-70℃ before use.

8. A method of preparing a coconut protein based fermented cheese containing phyllium seed oil according to claim 1, characterized in that, The amount of kumquat seed oil nanoemulsion added in step (2) is 1 wt.%-8 wt.% of the mixed matrix.

9. A method of preparing a coconut protein based fermented cheese containing vaccinium uliginosum seed oil according to claim 1, characterized in that, The amount of lactic acid bacteria starter added in step (3) is 0.25wt.%-1.5wt.%, the amount of CaCl2 added is 0.01wt.%-0.05wt.%, and the amount of rennet added is 0.1wt.%-1.0wt.%.

10. The coconut protein-based fermented cheese containing kumquat seed oil prepared by the method according to any one of claims 1-9.

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