Preparation method and application of BMFDS-loaded microcapsules

Abstract

The present invention discloses a method for preparing BMFDS-loaded microcapsules and their application. The preparation method comprises: mixing a mucin solution and a chitosan solution to undergo a complex coacervation reaction to obtain a composite wall material solution; the reaction pH is 7-9; the mass ratio of mucin to chitosan in the mixed solution is 3:1-1:3, and the total mass concentration of mucin and chitosan is 1.0%-2.0%; the reaction temperature is 20-40°C, and the heating time is 40-60 minutes; adding a BMFDS ethanol solution to the composite wall material solution at a BMFDS concentration of 0.002-0.05 mg / g, homogenizing, centrifuging, and freeze-drying the resulting precipitate to obtain the BMFDS-loaded microcapsules. The microcapsules are then added to a semi-finished plant-based meat product that has not been subjected to heating processing, and the finished plant-based meat product is obtained after baking. The BMFDS-loaded microcapsules, when added to the plant-based meat product, can significantly reduce the loss of exogenous aroma in the plant-based meat and enhance the flavor of the plant-based meat.

Description

Technical Field

[0001] The present invention belongs to the technical field of food processing, in particular to the technical field of plant meat and related additives, and specifically relates to a preparation method of BMFDS-loaded microcapsules and application thereof. Background Art

[0002] Meat flavorings are additives primarily based on meat flavors that improve the flavor and mouthfeel of foods. They are widely used in a variety of food industries, including meat products, frozen foods, convenience foods, catering ingredients, and baked goods. Existing meat flavorings primarily include mixed flavors and heat-reactive flavors. Basic meat aroma, characteristic meat aroma, roasted aroma, caramel aroma, onion-garlic aroma, smoky aroma, and spicy aroma are the primary contributors to meaty flavor. These aromas are produced by compounds such as alcohols, aldehydes, ketones, thiazoles, furans, pyrazines, and sulfur-containing compounds. Plant-based protein meats, made primarily from plant-based proteins such as soy protein, pea protein, and wheat protein, undergo pretreatment, seasoning, bonding, and molding processes. Their unique flavor and mouthfeel, along with their health and nutritional benefits, have gained increasing consumer attention in recent years. Consequently, the use of meat flavorings in plant-based meats is increasing. Research has shown that the addition of flavorings can effectively improve the flavor of plant-based meats, making them more similar to the aroma of authentic meat.

[0003] The aroma compounds in food have low molecular weight and high volatility. They are easy to evaporate and volatilize during processing and storage, causing huge losses, and also affect the odor of the spatial environment to a certain extent. Secondly, the aroma of the flavor is easily distorted by environmental influences such as temperature, humidity, oxygen, and pH. In addition, the flavor matrix is ​​rich in nutrients, which is conducive to the growth and reproduction of microorganisms, causing the flavor to spoil, which will limit the use and preservation of the flavor.

[0004] Microencapsulation technology is a micro-encapsulation protection technology that uses polymer materials to encapsulate unstable and volatile active substances. Microcapsules are microparticles made from these materials. Microencapsulation technology has been widely used in the food industry as an important food processing technology. This technology can change the volume and state of materials, solidifying them to facilitate storage and transportation. It can also improve the stability of the core material, isolate the influence of environmental factors such as temperature and light, and prevent oxidation and decomposition reactions of unstable components. It prevents the volatilization of aroma compounds and controls their release, controlling the release rate and duration of aroma compounds, thereby precisely controlling the release of aroma. Based on different principles, there are many technologies for preparing microcapsules, including spray drying, emulsification diffusion, complex coacervation, nanoprecipitation, layer-by-layer self-assembly, and other methods. Among them, complex coacervation has been widely used due to its mild reaction, simple preparation, good product stability, and high encapsulation rate.

[0005] In the preparation of microcapsules, three types of wall materials are used: lipids, proteins, and polysaccharides. Currently, gum arabic, gelatin, chitosan, sodium alginate, and soy protein are commonly used as wall materials in complex coacervation. However, in the food industry, particularly in the research and control of flavor in plant-based meats, there are still technical gaps in the application of microcapsules. More substances with potential as wall materials need to be discovered, and the preparation conditions for microcapsules require further optimization. Summary of the Invention

[0006] To overcome the shortcomings of the prior art, one objective of the present invention is to provide a method for preparing microcapsules loaded with bis(2-methyl-3-furyl) disulfide (BMFDS) using a mucin / chitosan composite coacervate as the wall material. This method addresses the problem of flavor dissipation during the processing of plant-based meat products and significantly enhances the sensory quality of plant-based meat. Another objective of the present invention is to provide a specific application method for the microcapsules. This objective is achieved through the following technical solutions:

[0007] In order to solve the above technical problems, the present invention provides a method for preparing BMFDS-loaded microcapsules, comprising the following steps:

[0008] 1) Weigh mucin (MUC) and chitosan (CS);

[0009] 2) preparing a mucin solution: dissolving the mucin in water to prepare a solution;

[0010] 3) preparing chitosan solution: dissolving chitosan in 1% acetic acid aqueous solution to prepare a solution;

[0011] 4) mixing the mucin solution and the chitosan solution for a complex coacervation reaction to obtain a composite wall material solution: the pH value of the complex coacervation reaction is 7-9; the mass ratio of mucin to chitosan in the mixed solution is 3:1-1:3; the total mass concentration of mucin and chitosan is 1.0%-2.0%; the reaction temperature after mixing the mucin solution and the chitosan solution is 20-40° C., and the heating time is 40-60 minutes;

[0012] 5) Preparation of bis(2-methyl-3-furyl) disulfide (BMFDS)-loaded microcapsules: A BMFDS ethanol solution was added to a composite wall material solution, wherein the concentration of BMFDS in the composite wall material solution was 0.002 to 0.05 mg / g. The mixture was homogenized and centrifuged at 10,000 rpm for 15 min at 4°C. The supernatant was discarded, and the precipitate (complex) obtained by centrifugation was freeze-dried to obtain the BMFDS-loaded microcapsules.

[0013] In a further embodiment, step 2) prepares a mucin solution by dissolving mucin in water to form a solution, magnetically stirring for 4 hours, and hydrating at 4°C overnight for use; step 3) prepares a chitosan solution by dissolving chitosan in a 1% acetic acid aqueous solution to form a solution, magnetically stirring for 4 hours, and hydrating at 4°C overnight; the mass concentration of the mucin solution prepared in step 2) and the chitosan solution prepared in step 3) are the same.

[0014] Furthermore, in step 4), the pH value of the complex coacervation reaction is 7.

[0015] In step 4), the mass ratio of mucin to chitosan in the mixed solution is 1:3.

[0016] In step 4), the total mass concentration of mucin and chitosan in the mixed solution is 2.0%.

[0017] In step 4), the reaction temperature after mixing mucin and chitosan is 30°C.

[0018] In step 4), the mucin solution and the chitosan solution are mixed and heated for 40 minutes.

[0019] Furthermore, in step 5), the concentration of BMFDS in the composite wall material solution is 0.01 mg / g.

[0020] The application of the BMFDS-loaded microcapsules is to add the microcapsules to a semi-finished plant meat product that has not been heated, and then bake the semi-finished plant meat product to obtain the finished plant meat product.

[0021] A further solution is to add the microcapsules to an aqueous solution of soy textured protein and fibronectin that is continuously stirred at 0-4°C for 12 hours so that the final flavor addition amount reaches 0.01 mg / g, and the mass ratio of fibronectin to soy textured protein is 2:8. After stirring and mixing, the mixture is placed in a mold for forming, and the baking temperature is 160-200°C and the baking time is 12-16 minutes to obtain the finished plant meat product.

[0022] Beneficial effects of the present invention:

[0023] (1) In the BMFDS-loaded microcapsules of the present invention, mucin and chitosan are used for complex coacervation and BMFDS is embedded. Mucin can form a complex with chitosan through non-covalent interactions (e.g., electrostatic interactions, hydrogen bonds, and hydrophobic interactions). By adjusting the mucin / chitosan solution ratio, total wall material concentration, pH, reaction time, and reaction temperature, microcapsules with the ability to significantly improve the stability of aroma compounds are prepared.

[0024] (2) The BMFDS-loaded microcapsules described in the present invention can significantly reduce the loss of exogenous aroma of plant meat and enhance the flavor of plant meat after being added to plant meat products. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The present invention will be further described below with reference to the accompanying drawings and examples.

[0026] Figure 1 Figure 1 shows the equilibrium phase turbidity and coacervate phase yield measured in Experiments 1-1 to 1-5. It also shows the effects of pH (A), MUC / CS ratio (B), total wall material concentration (C), reaction temperature (D), and reaction time (E) on the complex coacervation reaction between MUC and CS. Different lowercase letters indicate significant differences (P < 0.05).

[0027] Figure 2 These are infrared spectra of MUC, CS, and the MUC-CS complex coacervate prepared with a MUC / CS ratio of 1:3 in Experimental Example 1-2.

[0028] Figure 3 It is the DSC thermogram of MUC, CS and the MUC-CS complex coacervate prepared in the MUC / CS ratio of 1:3 in Experimental Example 1-2.

[0029] Figure 4 These are scanning electron micrographs of MUC (A), CS (B), and the MUC-CS complex coacervate prepared with a MUC / CS ratio of 1:3 in Experimental Example 1-2 (C).

[0030] Figure 5 Figure 2 shows the effects of adding MUC-CS-BMFDS microcapsules and SPI-CS-BMFDS microcapsules, prepared in Example 1-2 and Comparative Example 2-2, respectively, to the vegetable meat during processing at different baking temperatures (A) and different baking times (B). Different lowercase letters in the figure indicate significant differences (P < 0.05). DETAILED DESCRIPTION

[0031] Test Example 1-1

[0032] This test example provides a method for preparing a mucin and chitosan complex coacervation, the preparation method comprising the following steps:

[0033] (1) Weigh a certain amount of mucin and chitosan;

[0034] (2) Preparation of mucin solution: Dissolve mucin in water to a 2% solution, stir magnetically for 4 h, and store at 4°C overnight;

[0035] (3) Preparation of chitosan solution: Dissolve chitosan in 1% acetic acid aqueous solution to a 2% mass concentration solution, stir magnetically for 4 h, and store at 4°C overnight;

[0036] (4) Mixing the solutions to obtain a composite wall material solution: The mass ratio of MUC and CS was fixed at 1:1, and the total mass concentration of the wall materials (mucin and chitosan) was 2%. The two solutions were mixed in a 30°C water bath, and the pH value of the system was adjusted to 5, 6, 7, 8, and 9 using HCl and NaOH, and the stirring was continued for 40 min.

[0037] (5) Centrifuge at 10,000 rpm for 15 min at 4°C, measure the turbidity of the supernatant, and freeze-dry the complex to obtain the MUC-CS complex.

[0038] The results of equilibrium phase turbidity and coacervate phase yield during the preparation of MUC-CS complex are shown in Figure 2. Figure 1 As shown in A.

[0039] The mucin described in this example was purchased from Sigma-Aldrich Company, USA, with the product model number M2378; chitosan was purchased from Shanghai Yuanye Biotechnology Co., Ltd., with the product model number S11064.

[0040] Test Example 1-2

[0041] This test example provides a preparation method for mucin and chitosan complex coacervation. The raw materials used are the same as those in Test Example 1-1. The preparation method differs from that in Test Example 1-1 in that:

[0042] In step (4), the total mass concentration of the fixed wall material was 2%, and the mass ratios of MUC and CS were 5:1, 3:1, 1:1, 1:3, and 1:5, respectively. The two solutions were mixed in a 30°C water bath, and the pH value of the system was adjusted to 7 with HCl and NaOH, and stirring was continued for 40 minutes.

[0043] The results of equilibrium phase turbidity and coacervate phase yield during the preparation of MUC-CS complex are shown in Figure 2. Figure 1 As shown in B.

[0044] Test Examples 1-3

[0045] This test example provides a preparation method for mucin and chitosan complex coacervation. The raw materials used are the same as those in Test Example 1-1. The preparation method differs from that in Test Example 1-1 in that:

[0046] In step (4), a mass ratio of MUC to CS was selected as 1:1, and solutions with total wall material concentrations of 1.0%, 1.5%, 2.0%, 2.5%, and 3.0% were prepared, respectively. The two solutions were mixed in a 30°C water bath, and the pH value of the system was adjusted to 7 with HCl and NaOH, and stirring was continued for 40 minutes.

[0047] The results of equilibrium phase turbidity and coacervate phase yield during the preparation of MUC-CS complex are shown in Figure 2. Figure 1 As shown in C.

[0048] Test Examples 1-4

[0049] This test example provides a preparation method for mucin and chitosan complex coacervation. The raw materials used are the same as those in Test Example 1-1. The preparation method differs from that in Test Example 1-1 in that:

[0050] In step (4), the mass ratio of MUC and CS was fixed at 1:1, the total mass concentration of the wall material was 2%, and the two solutions were mixed in a water bath at 20°C, 30°C, 40°C, 50°C, and 60°C, respectively. The pH value of the system was adjusted to 7 with HCl and NaOH, and stirring was continued for 40 minutes.

[0051] The results of equilibrium phase turbidity and coacervate phase yield during the preparation of MUC-CS complex are shown in Figure 2. Figure 1 As shown in D.

[0052] Test Examples 1-5

[0053] This test example provides a preparation method for mucin and chitosan complex coacervation. The raw materials used are the same as those in Test Example 1-1. The preparation method differs from that in Test Example 1-1 in that:

[0054] In step (4), the mass ratio of MUC and CS was fixed at 1:1, the total mass concentration of the wall material was 2%, the two solutions were mixed in a 30°C water bath, the pH value of the system was adjusted to 7 with HCl and NaOH, and stirring was continued for 20 min, 30 min, 40 min, 50 min, and 60 min.

[0055] The results of equilibrium phase turbidity and coacervate phase yield during the preparation of MUC-CS complex are shown in Figure 2. Figure 1 As shown in E.

[0056] Experiment 1: Optimization of complex coacervation conditions

[0057] The experimental results are as follows Figure 1As described in AE, the results show that as the pH increases from 5.0 to 9.0, the equilibrium phase turbidity first decreases and then increases, while the coacervate yield first increases and then decreases. At a pH of 7.0, the equilibrium phase turbidity is lowest and the coacervate yield is highest. This indicates that the positive and negative charge densities of the biopolymers reach stoichiometric equilibrium, and the resulting complexes are stable under this pH condition. As the MUC / CS ratio decreases, the turbidity first decreases and then increases, while the coacervate yield first increases and then decreases. A ratio of 1:3 achieves the lowest turbidity and the highest coacervate yield. This is because anions and cations are not in the optimal mixing ratio, resulting in an excess of one component and a deficiency of the other, leading to an excess of polymer molecules in the equilibrium phase. A combined MUC and CS concentration of 2% results in the lowest equilibrium phase turbidity and the highest coacervate yield. At an appropriate total wall material concentration, the macromolecular backbone expands less, resulting in greater flexibility and easier interaction between charge sites, leading to complex coacervation. The equilibrium phase turbidity is lowest and the coacervate yield is highest at 30°C. In addition to electrostatic interactions, other interactions also occur during the complex coacervation process. At lower temperatures, more hydrogen bonds form during the complex coacervation process, while at higher temperatures, the high energy of the molecules hinders hydrogen bond formation. Furthermore, at high temperatures, proteins and polysaccharides swell and form adhesions. At 40 minutes, the equilibrium phase had the lowest turbidity and the highest yield of the coacervate phase, indicating that the reaction between MUC and CS was almost complete after 40 minutes of reaction.

[0058] Experiment 2, IR spectroscopy analysis, DSC analysis, and SEM morphology of MUC-CS complex

[0059] Appropriate amounts of MUC, CS, and MUC-CS freeze-dried powder (samples obtained by optimizing complex coacervation conditions, equivalent to the mass ratio of MUC and CS in Experimental Example 1-2 of 1:3) were mixed with KBr at a ratio of 1:100, ground in a mortar, and then compressed into tablets using a tablet press. The results were analyzed using an Avater.370 Fourier transform infrared spectrometer at 4000-400 -1 The sample was scanned within the scanning range with a KBr flake as the background. The spectrum was obtained after averaging 32 scans.

[0060] Accurately weigh 5 mg of MUC, CS, and MUC-CS complex samples (samples obtained by optimizing complex coacervation conditions) and seal them in an aluminum pan. Use a differential scanning calorimeter to scan the heat flow changes of the samples in the range of 20-220°C at a heating rate of 10°C / min.

[0061] Appropriate amounts of MUC, CS, and MUC-CS complex samples (samples obtained by optimizing complex coagulation conditions) were weighed, fixed on a sample stage with conductive tape, and then treated with gold spraying and photographed with a scanning electron microscope to observe the microscopic morphology of the samples.

[0062] Depend on Figure 2It can be seen that the infrared spectrum of MUC-CS has a characteristic absorption peak wavelength of -COOR group shifted to 1155 cm-1 relative to that of MUC. -1 , compared to CS, 1597cm -1 -NH 3+ The absorption peak of MUC and CS disappears, proving that there is electrostatic interaction between MUC and CS, which leads to complex coagulation. The spectral curves of the remaining positions in MUC-CS are similar to the superposition of the infrared spectral curves of MUC and CS. -1 A broader peak appeared, which was due to the overlap of -OH in its structure. After complex coagulation, the absorption peak intensity and wavelength of -OH changed, which indicated that there was still hydrogen bonding between MUC and CS.

[0063] Depend on Figure 3 As can be seen, the peak temperature of MUC is 145°C, while the peak temperature of CS is 128°C. This is due to their different structures and also indicates that MUC is more stable. The peak temperature of the complex coacervation product of MUC and CS is 141°C, and the transition temperature is higher than that of CS, indicating that a more stable structure has been formed between the two. The addition of protein enhances the thermal stability of CS.

[0064] Depend on Figure 4 It can be seen that MUC is an irregular aggregate, CS is a flat block structure, and the product obtained by the complex coagulation of MUC and CS is a loose and porous network structure, indicating that the negatively charged MUC is attached to the positively charged CS surface through electrostatic interaction to form a complex with a new structure, but MUC and CS are uniformly cross-linked in the liquid and have not yet embedded the core material, and finally a network structure is formed.

[0065] Example 1-1

[0066] A method for preparing BMFDS-loaded microcapsules comprises the following steps:

[0067] 1) Weigh a certain amount of mucin (MUC) and chitosan (CS);

[0068] 2) Preparation of mucin solution: Dissolve mucin in water to a 2% solution, stir magnetically for 4 h, and store at 4°C overnight;

[0069] 3) Preparation of chitosan solution: Dissolve chitosan in 1% acetic acid solution to a 2% solution, stir magnetically for 4 h, and store at 4°C overnight;

[0070] 4) The MUC solution and the CS solution were then mixed to obtain a composite wall material solution. The MUC and CS solutions were fixed at a 1:3 mass ratio and a total wall material (mucin and chitosan) concentration of 2%. The two solutions were mixed in a 30°C water bath. The pH of the system was adjusted to 7 with HCl and NaOH, and stirring was continued for 40 minutes.

[0071] 5) Preparation of BMFDS-loaded microcapsules: Dissolve an appropriate amount of BMFDS in anhydrous ethanol to obtain a BMFDS ethanol solution. Add an appropriate amount of the BMFDS ethanol solution to the composite wall material solution, achieving a BMFDS concentration of 0.002 mg / g. Centrifuge at 10,000 rpm for 15 minutes at 4°C, and discard the supernatant. Freeze-dry the resulting precipitate to obtain BMFDS microcapsules.

[0072] Example 1-2

[0073] This embodiment makes the following changes relative to embodiment 1-1:

[0074] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the mass concentration of BMFDS in the composite wall material solution was 0.01 mg / g. The rest of the steps were the same as those in Example 1-1.

[0075] Examples 1-3

[0076] This embodiment makes the following changes relative to embodiment 1-1:

[0077] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the mass concentration of BMFDS in the composite wall material solution was 0.05 mg / g. The rest of the steps were the same as those in Example 1-1.

[0078] Comparative Example 1-1

[0079] This comparative example is modified as follows relative to Example 1-1:

[0080] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the concentration of BMFDS in the composite wall material solution was 0.25 mg / g. The rest of the steps were the same as those in Example 1-1.

[0081] Comparative Example 2-1:

[0082] When the concentration of BMFDS in the composite solution was 0.002 mg / g, SPI / CS complex coacervates prepared BMFDS-loaded microcapsules.

[0083] The wall material is composed of soy protein isolate (SPI) and chitosan (CS). The specific steps are as follows:

[0084] 1) Weigh a certain amount of soy protein isolate and chitosan;

[0085] 2) Preparation of soy protein isolate solution: Dissolve soy protein isolate in water to a concentration of 2.67%, stir magnetically for 4 h, and incubate at 4°C overnight;

[0086] 3) Preparation of chitosan solution: Dissolve chitosan in 1% acetic acid solution to a 1% mass concentration, stir magnetically for 4 h, and store at 4°C overnight;

[0087] 4) The solutions were then mixed to obtain a composite wall material solution: the mass ratio of SPI to CS was fixed at 4:1, and the total concentration of the wall materials (SPI and CS) was 2%. The two solutions were mixed in a 25°C water bath. The pH of the system was adjusted to 6.5 with HCl and NaOH, and stirring was continued for 40 minutes.

[0088] The optimal conditions for complex coacervation of SPI and CS were determined according to the reference Huang G, Sun Y, Xiao J, et al. Complex coacervation of soybean protein isolate and chitosan[J]. Food Chemistry, 2012, 135(2): 534-539.

[0089] 5) Preparation of BMFDS-loaded microcapsules: Dissolve an appropriate amount of BMFDS in anhydrous ethanol to obtain a BMFDS ethanol solution; add an appropriate amount of the BMFDS ethanol solution to the composite wall material solution, wherein the BMFDS concentration in the composite wall material solution is 0.002 mg / g; centrifuge at 10,000 rpm for 15 min at 4°C, discard the supernatant, and freeze-dry the precipitate obtained by centrifugation to obtain SPI-CS-BMFDS microcapsules.

[0090] Comparative Example 2-2:

[0091] This comparative example is modified as follows relative to comparative example 2-1:

[0092] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the concentration of BMFDS in the composite wall material solution was 0.01 mg / g. The rest of the steps were the same as those in Comparative Example 2-1.

[0093] Comparative Examples 2-3:

[0094] This comparative example is modified as follows relative to comparative example 2-1:

[0095] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the concentration of BMFDS in the composite wall material solution was 0.05 mg / g. The rest of the steps were the same as those in Comparative Example 2-1.

[0096] Comparative Examples 2-4:

[0097] This comparative example is modified as follows relative to comparative example 2-1:

[0098] An appropriate amount of BMFDS ethanol solution was added to the composite wall material solution, wherein the concentration of BMFDS in the composite wall material solution was 0.25 mg / g. The rest of the steps were the same as those in Comparative Example 2-1.

[0099] Experiment 3: Determination of the embedding efficiency of BMFDS in microcapsules

[0100] Drawing of BMFDS standard curve: The characteristic absorption peak of BMFDS ethanol solution is 263nm, obtained by ultraviolet scanning spectrum at 200-700nm. Then the absorbance of BMFDS ethanol solution of different concentrations is measured to obtain the standard curve of BMFDS. The standard curve is y=24.788x-0.0083, R 2 =0.9999.

[0101] The microcapsule powders prepared in Examples 1-1 to 1-3 and Comparative Examples 1-1 and 2-1 to 2-4 were mixed with ethanol, centrifuged, and the supernatant collected. The BMFDS absorbance at 263 nm was measured to calculate the free BMFDS content. The same amount of microcapsule powder was then mixed with ethanol and sonicated at 60°C and 40 kHz for 30 minutes to extract the encapsulated BMFDS. The supernatant was collected by centrifugation and the absorbance was measured at the same wavelength. The total BMFDS content was calculated using a BMFDS standard curve. The results are shown in Table 1.

[0102] The BMFDS embedding efficiency is calculated as follows:

[0103]

[0104] Note: The total amount of BMFDS refers to the total mass of BMFDS in the composite solution, and the free BMFDS refers to the mass of BMFDS attached to the microcapsule powder. The difference between them is the mass of BMFDS embedded in the microcapsules.

[0105] Table 1

[0106]

[0107] With increasing BMFDS content, the encapsulation efficiency of MUC-CS microcapsules initially increased and then decreased, reaching 87.69% at a BMFDS content of 0.01 mg / g. The encapsulation efficiency of SPI-CS microcapsules for BMFDS decreased with increasing BMFDS content, reaching a maximum of 49.29%. A comparison revealed that the complex coacervation of MUC and CS effectively encapsulated BMFDS, surpassing that of the complex coacervation of SPI and CS.

[0108] Experiment 4: Application of BMFDS microcapsules in plant-based meat

[0109] The plant-based meat preparation method is based on the experimental method optimized by the research team in the early stage. Fibroblasted protein and textured soy protein were rehydrated by soaking them in water at a mass ratio of 2:8 and stirring continuously for 12 hours. BMFDS stock solution, MUC-CS-BMFDS microcapsules, and SPI-CS-BMFDS microcapsules were added to the rehydrated protein and stirred thoroughly. After mixing, the mixture was formed into patties with a thickness of 1.5 cm and a radius of 5 cm. Plant-based meat patties were prepared at different baking temperatures (flavoring added at 0.01 mg / g, baking time of 12 minutes, baking temperatures of 160°C, 180°C, and 200°C) and different baking times (flavoring added at 0.01 mg / g, baking temperature of 180°C, and baking times of 12 minutes, 14 minutes, and 16 minutes).

[0110] Extract aroma compounds from plant-based patties. Accurately weigh 2.0 g of sample and place in a sealed headspace vial. After equilibration at 55°C for 15 minutes, use an SPME fiber to adsorb volatile BMFDS in the headspace vial at 55°C for 40 minutes. Then, desorb at 250°C for 5 minutes to release the volatile compounds.

[0111] The preparation method of plant meat refers to Li Yan, Zeng Xiangquan, Du Wenbin, et al. Effects of different vegetable oils on the flavor and sensory properties of plant meat [J]. Journal of Food Science and Technology, 2022, 40(02): 31-44.

[0112] The extraction method of aroma compounds in plant patties was referenced from: Li X, Zeng X, Xi Y, et al. Effects of non-covalent interactions between pectin and volatile compounds on the flavor release of tomato paste[J]. Food Hydrocolloids, 2022, 133: 107886. and some modifications were made.

[0113] The effects of different baking temperatures and different baking times on the flavor of plant meat are as follows: Figure 5 As shown in (AB).

[0114] according to Figure 5A. As the baking temperature increased from 160°C to 200°C, the BMFDS content in the plant meat decreased significantly. In the control group, the aroma compounds decreased by 41.03%. The BMFDS content in the MUC-CS and SPI-CS groups was lower than that in the control group (the control group used BMFDS solution, and the initial concentration of BMFDS in the three groups was 0.01 mg / g. The encapsulation rates of BMFDS in the MUC-CS and SPI-CS groups were 87.69% and 40.5%, respectively). However, the aroma loss during the baking temperature change was only 19.32%, and the aroma loss in the SPI-CS group was only 21.52%. This shows that encapsulation can protect the aroma compounds, reduce the aroma loss during processing, and have a better sustained-release effect.

[0115] according to Figure 5 B. As the roasting time increased from 12 to 14 minutes, the BMFDS content in the plant-based meat decreased significantly. In the control group, the aroma compounds decreased by 42.45%. The aroma loss of BMFDS in the MUC-CS and SPI-CS groups with increased roasting time was only 20.03%, while the aroma loss in the SPI-CS group was only 24.53%. This indicates that encapsulating BMFDS effectively reduces aroma loss in plant-based meat, and that MUC not only has a higher BMFDS encapsulation rate but also has a better protection effect than SPI.

[0116] The experiments described above primarily compared the retention of BMFDS in plant-based meats as baking temperature increased. As can be seen, the aroma loss of the encapsulated flavor during the temperature change was significantly reduced compared to the control group. This suggests that encapsulation protects aroma compounds, reducing aroma loss during processing and resulting in a better sustained-release effect.

[0117] In summary, mucin and chitosan coagulation was optimal at a pH of 7.0, a MUC / CS ratio of 1:3, a total wall material concentration of 2%, a temperature of 30°C, and a reaction time of 40 min. The formation of a complex coacervate between MUC and CS was primarily through electrostatic interactions. This complex exhibited a gel-like network structure and enhanced the thermal stability of CS. The highest encapsulation efficiency, 87.69%, was achieved at a BMFDS content of 0.01 mg / g. However, the encapsulation efficiency of BMFDS by complex coacervation of SPI and CS was significantly weaker than that of MUC and CS. During the processing of plant-based meat, both baking temperature and time significantly affect the content of aroma compounds in the meat. Increased baking temperature and time resulted in varying degrees of BMFDS loss. However, encapsulation of BMFDS effectively reduced aroma loss, and the protective effect of complex coacervation of MUC and CS on BMFDS was superior to that of SPI.

[0118] Finally, it should be noted that the above is only used to illustrate the technical solution of the present invention and is not limiting. Although the present invention is described in detail with reference to the preferred arrangement scheme, ordinary technicians in this field should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. Application of BMFDS-loaded microcapsules, characterized in that: The microcapsules are added to a semi-finished plant meat product that has not been heated, and the finished plant meat product is obtained after baking. The microcapsules are added to an aqueous solution of soy textured protein and fibronectin that has been stirred continuously for 12 hours at 0-4°C so that the final BMFDS addition amount reaches 0.01 mg / g, and the mass ratio of fibronectin to soy textured protein is 2:

8. After stirring and mixing, the mixture is placed in a mold for forming, and the baking temperature is 160-200°C and the baking time is 12-16 minutes to obtain the finished plant meat product. The preparation method of the microcapsules comprises the following steps: 1) Weigh mucin and chitosan; 2) Prepare mucin solution: dissolve mucin in water to prepare a solution; 3) Prepare chitosan solution: dissolve chitosan in 1% acetic acid aqueous solution to prepare a solution; 4) mixing the mucin solution and the chitosan solution for a complex coacervation reaction to obtain a composite wall material solution. The pH value of the complex coacervation reaction is 7-9. The mass ratio of mucin to chitosan in the mixed solution is 3:1-1:3, and the total mass concentration of mucin and chitosan is 1.0%-2.0%. The reaction temperature after mixing the mucin solution and the chitosan solution is 20-40°C, and the heating time is 40-60 minutes. 5) Preparation of bis(2-methyl-3-furyl) disulfide (BMFDS)-loaded microcapsules: A BMFDS ethanol solution was added to a composite wall material solution, where the concentration of BMFDS in the composite wall material solution was 0.002 to 0.05 mg / g. The mixture was homogenized and centrifuged at 10,000 rpm for 15 min at 4°C. The supernatant was discarded, and the precipitate obtained by centrifugation was freeze-dried to obtain the BMFDS-loaded microcapsules.

2. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: Step 2) prepare a mucin solution: dissolve mucin in water to form a solution, stir magnetically for 4 hours, and hydrate at 4°C overnight for use; Step 3) prepare a chitosan solution: dissolve chitosan in 1% acetic acid aqueous solution to form a solution, stir magnetically for 4 hours, and hydrate at 4°C overnight; the mass concentration of the mucin solution prepared in step 2) and the chitosan solution prepared in step 3) are the same.

3. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 4), the pH value of the complex coacervation reaction is 7.

4. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 4), the mass ratio of mucin to chitosan in the mixed solution is 1:

3.

5. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 4), the total mass concentration of mucin and chitosan in the mixed solution is 2.0%.

6. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 4), the reaction temperature after mixing mucin and chitosan is 30°C.

7. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 4), the mucin solution and the chitosan solution are mixed and heated for 40 minutes.

8. The use of the BMFDS-loaded microcapsules according to claim 1, characterized in that: In step 5), the concentration of BMFDS in the composite wall material solution is 0.01 mg / g.

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