A method for improving the color and digestibility of cell meat products and products thereof

Abstract

The present application relates to the technical field of food processing, and provides a method for improving color quality of cell meat products and improving their digestibility and products thereof. Through the synergistic effect of cell culture meat and lycopene, the color quality of cell meat products is significantly improved, the darkening of color and flavor deterioration caused by protein oxidation and lipid peroxidation during processing and storage are effectively inhibited, and the emulsion stability of the meat paste system and the integrity of the protein network structure are enhanced, thereby prolonging the shelf life on the basis of maintaining the original texture and taste of the products. Cell culture meat and lycopene can also synergistically improve the digestibility of meat products, improve the protein hydrolysis efficiency and amino acid bioavailability, and reduce the digestive burden. The present application provides a feasible technical path for the development of high-quality healthy meat products.

Description

Technical Field

[0001] This invention relates to the field of food processing technology, and in particular to a method and product for improving the color and digestibility of cell-based meat products. Background Technology

[0002] As consumers increasingly demand higher nutritional quality in food, the meat processing industry is facing a transformation towards functional and health-oriented products. Traditional meat processing typically involves chopping, heat treatment, and refrigeration. These processes easily induce protein and lipid oxidation, leading to quality problems such as discoloration, abnormal flavor, and deteriorated texture. Protein oxidation not only affects sensory appeal but can also cause protein restructuring, aggregation, and masking of digestive enzyme cleavage sites, ultimately reducing digestibility and amino acid bioavailability in the human gastrointestinal tract.

[0003] To address these issues, research on meat product quality improvement has gradually expanded from simply optimizing processing techniques to adding exogenous functional components. While traditional improvers such as phosphates, emulsifiers, and edible colloids have shown some effectiveness in water retention and texture regulation, the increasing consumer demand for clean labels and naturally derived ingredients has prompted researchers to focus on the application potential of plant-derived bioactive substances in meat products. Natural components such as plant polyphenols, flavonoids, and carotenoids, due to their antioxidant, antibacterial, and structural modification functions, are gradually becoming a hot topic in meat product quality control research.

[0004] In terms of improving the digestibility of meat products, existing research mainly focuses on improving protein hydrolysis sensitivity by adjusting processing parameters or adding enzymes. However, systematic studies on the regulation of protein structural stability and digestive behavior by natural active components in the complex matrix of meat products remain limited. The interaction mechanisms between active components and meat proteins, their stability changes during processing, and their ultimate impact on the distribution and functional activity of digestible products require further elucidation. A deeper understanding of these issues is crucial for developing novel meat products (such as cultured meat) that combine excellent sensory qualities with high digestibility and absorption efficiency. Summary of the Invention

[0005] To address the aforementioned problems in the prior art, this invention provides a method and product for improving the color and digestibility of cell-based meat products.

[0006] This invention achieves a dual improvement in meat product quality and digestibility through the synergistic effect of cell-cultured meat and lycopene. Cell-cultured meat is rich in bioactive proteins and unsaturated lipids, and its fresh tissue structure enhances the emulsification stability of the minced meat system, improving product texture. Lycopene, as a potent fat-soluble antioxidant, effectively scavenges free radicals induced by heat and oxygen during processing and storage, inhibits the oxidative aggregation of myofibril proteins, and maintains the integrity of protein structure. Stable protein structure helps retain digestive enzyme action sites, thereby improving the efficiency of protein hydrolysis in the gastrointestinal tract. The combined use of these two ingredients optimizes the sensory quality of meat products while enhancing their nutritional bioavailability.

[0007] In a first aspect, the present invention provides a method for improving the quality of meat products and enhancing their digestibility, the method comprising: adding cell-cultured meat and lycopene in a mass ratio of (90-98):1 to the raw materials of the meat products.

[0008] According to a method for improving the quality and / or digestibility of meat products provided by the present invention, preferably, the mass ratio of the cell-cultured meat to the lycopene is 90:1, 91:1, 92:1, 93:1, 94:1, 95:1, 96:1, 97:1 or 98:1.

[0009] More preferably, the mass ratio of the cell-cultured meat to the lycopene is 96:1.

[0010] According to a method for improving the quality and / or digestibility of meat products provided by the present invention, preferably, 4% wt to 5% wt of cell-cultured meat and / or 0.04% wt to 0.06% wt of lycopene are added to the raw materials of the meat products.

[0011] More preferably, 4%wt, 4.1%wt, 4.2%wt, 4.3%wt, 4.4%wt, 4.5%wt, 4.6%wt, 4.7%wt, 4.8%wt, 4.9%wt or 5%wt of cell-cultured meat are added to the raw materials of the meat product.

[0012] More preferably, 4.8% wt of cell-cultured meat is added to the raw material of the meat product.

[0013] More preferably, 0.040%wt, 0.045%wt, 0.050%wt, 0.055%wt or 0.06%wt of lycopene are added to the raw materials of the meat product.

[0014] More preferably, 0.050% wt lycopene is added to the raw material of the meat product.

[0015] According to the present invention, a method for improving the quality and / or digestibility of meat products is provided, wherein the raw materials of the meat products comprise animal meat and animal oil; the mass ratio of the cell-cultured meat to the animal meat is 3:(40-60), and / or the mass ratio of the lycopene to the animal oil is 1:(110-130).

[0016] More preferably, the mass ratio of the animal meat to the animal oil is (35-45):3.

[0017] More preferably, the mass ratio of the animal meat to the animal oil is 35:3, 36:3, 37:3, 38:3, 39:3, 40:3, 41:3, 42:3, 43:3, 44:3 or 45:3.

[0018] More preferably, the mass ratio of the animal meat to the animal oil is 40:3.

[0019] More preferably, the mass ratio of the cell-cultured meat to the animal meat is 3:40, 3:41, 3:42, 3:43, 3:44, 3:45, 3:46, 3:47, 3:48, 3:49, 3:50, 3:51, 3:52, 3:53, 3:54, 3:55, 3:56, 3:57, 3:58, 3:59, or 3:60.

[0020] More preferably, the mass ratio of the cell-cultured meat to the animal meat is 3:50.

[0021] More preferably, the mass ratio of the lycopene to the animal oil is 1:110, 1:115, 1:120, 1:125 or 1:130.

[0022] More preferably, the mass ratio of lycopene to animal oil is 1:120.

[0023] More preferably, the method includes: first mixing the lycopene with the animal oil, and then mixing the resulting mixture with other components in the raw materials to obtain minced meat.

[0024] When meat is high-speed stirred or pounded, friction generates heat, causing the minced meat temperature to rise. This not only damages the protein structure in the meat, affecting its final elasticity and texture, but also makes it easier for bacteria to grow. Adding ice cubes can effectively lower the temperature, ensuring the entire process is carried out at a low temperature. The low-temperature environment allows the minced meat to better absorb and lock in moisture, resulting in beef balls that are more tender and juicy inside, rather than dry. Furthermore, at low temperatures, the salt-soluble proteins in the meat can be extracted more effectively and form a stable and elastic gel network structure during subsequent heating, providing a bouncy texture.

[0025] More preferably, the method includes: mixing the raw materials of the meat product at a low temperature to obtain minced meat.

[0026] More preferably, the method includes: first mixing the lycopene with the animal oil, then mixing the resulting mixture with other components in the raw materials, and then adding ice; the mass ratio of the animal meat to the ice is (6-10):3.

[0027] More preferably, the mass ratio of the animal meat to the ice is 6:3, 7:3, 8:3, 9:3, or 10:3.

[0028] More preferably, the mass ratio of the animal meat to the ice is 8:3.

[0029] More preferably, the method further includes heating the minced meat.

[0030] More preferably, the heating method includes any one or more of steaming, boiling, frying, grilling, and baking.

[0031] According to a method for improving the quality and / or digestibility of meat products provided by the present invention, preferably, the cell-cultured meat includes cell-cultured meat of poultry origin.

[0032] More preferably, the cell-cultured meat is pigeon-derived cell-cultured meat.

[0033] More preferably, the cell-cultured meat comprises muscle tissue and adipose tissue obtained by differentiation and culture of muscle satellite cells.

[0034] More preferably, the mass ratio of the muscle tissue to the adipose tissue is (8-12):1.

[0035] More preferably, the mass ratio of the muscle tissue to the adipose tissue is 8:1, 9:1, 10:1, 11:1 or 12:1.

[0036] More preferably, the mass ratio of the muscle tissue to the adipose tissue is 10:1.

[0037] According to the present invention, a method for improving the quality and / or digestibility of meat products is provided, wherein the animal meat includes any one or more of beef, pork, mutton, fish, and chicken, and / or the animal oil includes any one or more of lard, butter, fish oil, and chicken oil.

[0038] More preferably, the animal meat is beef.

[0039] More preferably, the animal oil is lard.

[0040] More preferably, the animal meat is beef, and the animal fat is lard.

[0041] More preferably, the raw materials of the meat product further include: 1%wt to 5%wt salt, 0.5%wt to 0.7%wt sugar, 0.1%wt to 0.2%wt baking soda and 6%wt to 7%wt tapioca starch.

[0042] More preferably, the raw materials for the meat products further include: 1%wt, 2%wt, 3%wt, 4%wt or 5%wt salt.

[0043] More preferably, the raw materials for the meat product further include: 2% wt salt.

[0044] More preferably, the raw materials of the meat product further include: 0.50%wt, 0.52%wt, 0.54%wt, 0.56%wt, 0.58%wt, 0.60%wt, 0.62%wt, 0.64%wt, 0.66%wt, 0.68%wt or 0.70%wt sugar.

[0045] More preferably, the raw materials of the meat product further include: 0.64% wt sugar.

[0046] More preferably, the raw materials for the meat products further include: 0.10%wt, 0.12%wt, 0.14%wt, 0.16%wt, 0.18%wt or 0.20%wt of baking soda.

[0047] More preferably, the raw material of the meat product further includes: 0.16%wt baking soda.

[0048] More preferably, the raw materials for the meat products further include: 6.00%wt, 6.05%wt, 6.10%wt, 6.15%wt, 6.20%wt, 6.25%wt, 6.30%wt, 6.35%wt, 6.40%wt, 6.45%wt, 6.50%wt, 6.55%wt, 6.60%wt, 6.65%wt, 6.70%wt, 6.75%wt, 6.80%wt, 6.85%wt, 6.90%wt, 6.95%wt, or 7.00%wt of tapioca starch.

[0049] More preferably, the raw material of the meat product further includes: 6.35% wt tapioca starch.

[0050] According to the present invention, a method for improving the quality and / or digestibility of meat products is provided. Preferably, improving the quality of meat products includes any one of the following: (1) improving the sensory evaluation of meat products; (2) improving the texture of meat products; (3) inhibiting the oxidation of proteins and / or fats in meat products.

[0051] More preferably, improving the sensory evaluation of meat products includes improving any one or more of the following: color, texture, odor, taste, and overall acceptability of meat products.

[0052] According to the present invention, a method for improving the quality and / or digestibility of meat products is provided. Preferably, improving the digestibility of meat products includes any one of the following: (1) increasing the degree of hydrolysis of meat products; (2) increasing the surface hydrophobicity of meat products; (3) reducing the particle size of meat products.

[0053] On the other hand, the present invention also provides meat products prepared by any of the methods described.

[0054] According to the present invention, a method for improving the quality and / or digestibility of meat products is provided, wherein the meat products are preferably meatballs, meat patties, sausages or chunks of meat.

[0055] More preferably, the meat product is a beef ball.

[0056] Compared with the prior art, the present invention has the following beneficial effects: This invention, through the synergistic addition of cell-cultured meat and lycopene, comprehensively enhances the sensory quality and functional properties of meat products. Cell-cultured meat provides the system with structurally complete protein components, and combined with the strong antioxidant activity of lycopene, it significantly enhances the emulsification stability and protein network structure integrity of the meat paste system. This effectively inhibits color darkening and flavor deterioration caused by protein oxidation and lipid peroxidation during processing and storage, extending shelf life while maintaining the original texture and taste of the product. This synergistic effect also significantly improves the digestibility of meat products, increasing the hydrolysis efficiency of protein in the gastrointestinal tract and the bioavailability of amino acids, thus reducing the digestive burden. This invention provides a practical and feasible technical path for the development of high-quality, healthy meat products. Attached Figure Description

[0057] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0058] Figure 1 These are the appearance and cross-sectional photographs of M, LM, and LOHML provided in Test Example 1 of this invention.

[0059] Figure 2The apparent viscosity results of M, LM and LOHM provided in Test Example 1 of this invention are shown below; A is the storage modulus (G'); B is the loss modulus (G"); C is the tan δ value; D is the apparent viscosity curve of the three cell cultured beef meatball samples.

[0060] Figure 3 These are the TBARS values ​​of M, LM, and LOHM provided in Test Example 1 of this invention; different lowercase letters indicate significant differences between adding and not adding lycopene. P <0.05); different capital letters indicate significant differences between adding and not adding cultured meat (not added). P <0.05).

[0061] Figure 4 This refers to the protein oxidation of M, LM, and LOHM provided in Test Example 1 of this invention; A represents the statistical results of carbonyl content; B represents the statistical results of free thiol content.

[0062] Figure 5 This refers to the digestibility of M, LM, and LOHM provided in Test Example 2 of this invention; A represents the statistical results of the degree of hydrolysis; B represents the statistical results of the surface hydrophobicity. Detailed Implementation

[0063] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0064] Unless otherwise specified, the experimental methods involved in the following embodiments are conventional methods in the art. For example, you can refer to the experimental manual in the art or follow the conditions recommended in the manufacturer's instructions.

[0065] Unless otherwise specified, all experimental materials and reagents used in the following examples are commercially available.

[0066] The experiments described in the following examples were processed and statistically analyzed using Microsoft Excel 2016 and IBM SPSS 26.0. One-way ANOVA was used for one-way ANOVA, and Duncan's multiple comparison analysis was used for the data (P < 0.05). Unless otherwise specified, all experiments were repeated three times, and all data are expressed as mean ± standard deviation.

[0067] Example 1: Preparation of cell-cultured beef meatballs This embodiment uses beef and cell-cultured meat as the main raw materials to prepare cell-cultured meat beef balls.

[0068] 1. Preparation of cell-cultured meat Cell-cultured meat was prepared using the method described in the existing technology "Cultural optimization of pigeon muscle satellite cells and nutritional evaluation of the pigeon-derived cultivated meat" (10.1016 / j.jfutfo.2024.05.014). The specific method is as follows: Myosatellite cells (Pax7 positivity rate ≥95%) were isolated and purified from pigeon leg muscle tissue. After proliferation and expansion, the myosatellite cells were subjected to myogenic and adipogenic differentiation, respectively.

[0069] Myogenic differentiation was induced for 4–6 days using DMEM high-glucose medium containing 10% fetal bovine serum until clearly fused myotube structures were formed, and the cultured muscle tissue was collected. Adipogenic differentiation was induced for 7–10 days using McCoy's 5A medium containing 3% fetal bovine serum and 3% chicken serum until Oil Red O or BODIPY staining showed that the cells were filled with lipid droplets, and the cultured adipose tissue was collected.

[0070] Cell-cultured muscle tissue and adipose tissue are mixed at a mass ratio of 10:1 to obtain cell-cultured meat.

[0071] 2. Pre-treatment of beef After removing connective tissue and visible fat from fresh beef, it is ground.

[0072] 3. Preparation of meatballs Weigh each ingredient according to the recipe shown in Table 1, then add ice at a beef:ice mass ratio of 8:3 and mix thoroughly to obtain minced meat.

[0073] Table 1. Formula for cell-cultured beef meatballs

[0074] Squeeze the minced meat into meatballs by hand (weighing about 20 g and about 2 cm in diameter), put them into 80°C water and cook for 15 minutes to obtain cell cultured beef meatballs (denoted as M). After cooking, cool to room temperature and wipe off the surface moisture, then freeze at -18°C for later use.

[0075] Example 2: Preparation of cell-cultured beef meatballs This embodiment uses beef and cell-cultured meat as the main raw materials to prepare cell-cultured meat beef balls.

[0076] 1. Preparation of cell-cultured meat Same as Example 1.

[0077] 2. Pre-treatment of beef Same as Example 1.

[0078] 3. Preparation of meatballs Weigh each ingredient according to the formula shown in Table 2. Mix the lycopene directly with the lard and other ingredients evenly. Then add ice at a beef:ice mass ratio of 8:3 and beat to obtain minced meat.

[0079] Table 2. Formula for cell-cultured beef meatballs

[0080] Squeeze the minced meat into meatballs by hand (weighing about 20 g and about 2 cm in diameter), put them into 80°C water and cook for 15 minutes to obtain cell cultured beef meatballs (denoted as LM). After cooking, cool to room temperature and wipe off the surface moisture, then freeze at -18°C for later use.

[0081] Example 3: Preparation of cell-cultured beef meatballs This embodiment is basically the same as Embodiment 2, except that the method of adding lycopene in step "3, preparation of meatballs" is different, as follows: Weigh each ingredient according to the formula shown in Table 2. Then, homogenize the lycopene and lard at 10,000 r / min for 5 min to obtain a lycopene-lard mixture. Then mix the mixture with other ingredients. Finally, add ice at a beef:ice mass ratio of 8:3 and beat to obtain minced meat.

[0082] Squeeze the minced meat into meatballs by hand (weighing about 20 g and about 2 cm in diameter), put them into 80°C water and cook for 15 minutes to obtain cell cultured beef meatballs (denoted as LOHM). After cooking, cool to room temperature and wipe off the surface moisture, then freeze at -18°C for later use.

[0083] Example 4: Preparation of Beef Balls This embodiment uses beef as the main ingredient to prepare beef balls, without adding cell-cultured meat.

[0084] 1. Pre-treatment of beef Same as Example 1.

[0085] 2. Preparation of meatballs Weigh each ingredient according to the formula shown in Table 3. First, homogenize the lycopene and lard at 10,000 r / min for 5 min to obtain a mixture of lycopene and lard. Then, mix the mixture with the other ingredients.

[0086] Table 3. Recipe for Beef Balls

[0087] Squeeze the minced meat into meatballs by hand (weighing about 20 g and about 2 cm in diameter), put them into 80℃ water and cook for 15 minutes to get beef meatballs (denoted as L). After cooking, cool to room temperature and wipe off the surface moisture, then freeze at -18℃ for later use.

[0088] Test Example 1: The effect of lycopene addition method on the quality of cell-cultured beef meatballs This embodiment evaluates the quality of M, LM, LOHM and L prepared in Examples 1 to 4 from the perspectives of pH, sensory evaluation, texture, gel strength, shear force, rheological properties, lipid oxidation and protein oxidation.

[0089] 1. Effect on pH (1) Test method The pH values ​​of M, LM, LOHM, and L were determined according to the method described in "Acorn (Quercus ithaburensis) Flour's effect on the physicochemical, textural, and sensory characteristics of raw and cooked beefmeatballs" (DOI: 10.1016 / j.ijgfs.2024.100887). The specific method is as follows: A 3 g beef meatball sample was homogenized with 30 mL of distilled water, and the pH value was measured using a digital pH meter. The pH meter was standardized using standard solutions with pH values ​​of 4.01, 6.86, and 9.18 before pH measurement.

[0090] (2) Test results The test results showed that different treatments all had a certain impact on pH value, and this impact showed a consistent trend before and after heating. In the minced meat stage, the pH values ​​of groups M and LM were higher, while the pH values ​​of groups LOHM and L decreased significantly. Therefore, homogenization treatment was the main factor affecting pH.

[0091] The overall pH of the cell-cultured meatballs was higher after cooking than before cooking, indicating that protein denaturation and water evaporation during heating may have exposed weakly basic groups, leading to an overall increase in pH. Despite this, the LOHM and L groups maintained the lowest pH values ​​in the final product, significantly lower than the M and LM groups. This suggests that the emulsion structure formed after homogenization may cause lycopene or lipids to release more acidic groups, or accelerate proteolysis to produce free amino groups, thereby lowering the system pH. The lower pH value of the LOHM group is closer to the isoelectric point of muscle proteins, which may have a synergistic effect on subsequent gel formation, texture construction, and oxidation processes.

[0092] 2. Impact on sensory evaluation (1) Test method Referring to the nine-point scoring system of "Color, sensory and textural attributes of beef frankfurter, beefham and meat-free sausage containing tomato pomace" (DOI: 10.1016 / j.meatsci.2014.03.017), the sensory characteristics of three types of cell-cultured beef meatballs (M, LM, LOHM) and cell-free cultured meat (L) were determined. The specific method is as follows: Ten sensory evaluators (including 5 men and 5 women, aged between 23 and 30 years, professionals trained in sensory evaluation) used a nine-point scoring system (1 = very unsatisfactory, 9 = very satisfactory) to evaluate the color, texture, odor, taste, and overall acceptability of the cell-cultured beef meatballs.

[0093] (2) Test results Table 4. Effects of lycopene addition method on sensory evaluation of cell-cultured beef meatballs

[0094] Note: Different lowercase letters indicate significant differences. P <0.05).

[0095] like Figure 1 As shown, compared to M, LM and LOHM are significantly redder in color due to the addition of lycopene. L and LOHM have essentially the same appearance.

[0096] The sensory evaluation of beef meatballs showed significant differences depending on the method of lycopene addition. As shown in Table 4, the scores for color, texture, aroma, taste, and overall acceptability of beef meatballs from LM and LOHM were significantly higher than those from M. There were no significant differences in texture and taste among the three types of cell-cultured meat beef meatballs. This indicates that adding lycopene can significantly improve the multi-dimensional sensory experience of color, aroma, and taste of cell-cultured meat meatballs while maintaining their original texture and taste. However, L, due to the absence of cell-cultured meat, showed a significant decrease in texture, taste, and overall acceptability compared to LOHM.

[0097] 3. Effects on texture, gel strength, and shear force (1) Test method Three types of cell-cultured beef meatballs were cut into cylinders with a diameter of 2.0 cm and a height of 2.0 cm using a sampler. Then, a texture analyzer was used to test the texture, gel strength, and shear force, respectively.

[0098] Texture determination: Texture profile analysis mode was used; Probe model: P / 36 R flat-bottomed cylindrical probe; Measurement conditions were as follows: Pre-measurement rate 2 mm / s; Test rate 5 mm / s; Post-measurement rate 5 mm / s; Compression degree 50%; Residence time between two compressions 5 s; Trigger type automatic; Trigger force 10 g.

[0099] Determination of gel strength: The puncture mode was used, where the gel strength was the product of the breaking force and the indentation distance; the probe model was P / 0.5 HS, the pre-test rate was 2 mm / s, the test rate was 1 mm / s, the post-test rate was 2 mm / s, the shearing degree was 10 mm, the triggering type was automatic, and the triggering force was 10 g.

[0100] Shearing force: Using a machine equipped with a V-shaped cutting blade, the meat column is sheared along the vertical direction of the cylinder, the shearing force value is recorded, and the average value is calculated.

[0101] (2) Test results Table 5. Effects of lycopene addition method on texture, gel strength, and shear force of cell-cultured beef meatballs

[0102] Note: Different lowercase letters indicate significant differences. P <0.05).

[0103] The method of adding lycopene has a significant regulatory effect on textural properties. As shown in Table 5, LM has the highest hardness, significantly higher than M, indicating that lycopene co-solubility with lipids can promote the stable binding of lipid droplets and protein networks, constructing a dense structure. In contrast, LOHM has the lowest hardness, suggesting that although homogenization enhances lipid droplet dispersion, it may weaken network stability due to excessively fine particle size or shear damage to the protein structure. LOHM's cohesiveness and elasticity indices are significantly lower than M and LM, indicating that its internal structure is loose and has poor resilience. LM and LOHM have significantly higher adhesiveness than M, with LOHM having higher adhesiveness than LM, presumably because homogenization causes protein adsorption on the lipid droplet surface, enhancing the sample interfacial viscosity. LOHM reaches the highest gel strength, but its shear and chewiness are relatively low, indicating that while physical emulsification enhances structural density, it does not effectively maintain its overall textural properties. LM has the highest shear force, while the LOHM group has the lowest, indicating that its structure is more prone to breakage.

[0104] LOM achieves uniform lycopene release and lipoprotein synergistic enhancement through an "oil-soluble + heterogeneous" approach, significantly improving structural stability while also considering sensory chewing properties. While LOHM exhibits higher apparent adhesiveness, its softening tendency makes it suitable for developing soft, low-chew-burden products.

[0105] Because L did not contain cell-cultured meat, its gel strength was significantly lower and its adhesiveness was higher compared to LOHM.

[0106] 4. Effects on rheological properties (1) Test method Following the method described in "Catechins affect the oil-holding capacity of meat batters by changing the structure and emulsifying properties of surface proteins at the fat globules" (DOI: 10.1016 / j.ijbiomac.2023.126474), the dynamic rheology of chyme samples from three cell-cultured beef meatballs (M, LM, and LOHM) was detected. The specific methods are as follows: The instrument was calibrated before testing, with a gap temperature compensation of 20℃–80℃. A 40 mm plate clamp was used, and the chopped chyme sample was evenly spread on the center of the sample stage. The clamp was sealed with silicone oil around the perimeter to prevent air contact. The gap between the upper and lower plates was 0.8 mm, the strain was 0.05%, and the heating program was selected, increasing from 25℃ to 85℃ at a heating rate of 2℃ / min. Continuous shearing was performed at an oscillation frequency of 1 Hz. G' (storage modulus) and G'' (loss modulus) were measured.

[0107] Static rheology: A rheometer was used with a 40 mm plate for testing. The temperature was set to 25°C, ramp mode was selected, and the shear rate was 0.1–100 s⁻¹. -1 The test results were measured by the change in viscosity during the shear rate variation process. The experiment was repeated three times. Data were plotted on a logarithmic scale based on apparent viscosity as a function of shear rate.

[0108] (2) Test results like Figure 2 As shown in Figure A, the storage modulus (G') changes in three stages with increasing temperature. In the initial stage (25℃~55℃), myosin light chains are activated, a weakly elastic network is formed, and G' increases slowly. From 55℃ to 65℃, the protein structure unwinds, causing the network to relax, and G' decreases slightly. From 65℃ to 85℃, actin crosslinks with denatured myosin, and G' increases rapidly. The LOM group has the highest G' throughout the process because lycopene is uniformly dispersed, which enhances the protein-lipid interface interaction. The LOHM group has a lower G' in the early stage due to the disruption of network continuity caused by homogenization, but it recovers in the later stage due to the accelerated crosslinking of nanoemulsion droplets, showing a characteristic of first decreasing and then increasing.

[0109] like Figure 2 As shown in Figure B, the loss modulus (G'') and the G' trend are synergistic, reflecting the viscous dissipation capacity. In the low-temperature region (25℃~55℃), increased molecular chain friction leads to an increase in G'', with the LOHM group having the lowest G'' due to the nanoemulsion droplet interface effect; in the high-temperature region (65℃~85℃), protein cross-linking drives a rapid increase in G'', with the OHM group having a significantly lower G'' than other groups due to oxidation causing oil-water phase separation.

[0110] like Figure 2 As shown in C, the loss tangent (tan δ) characterizes the viscoelastic equilibrium. In the low-temperature region, viscosity dominates, and tan δ increases. The LOHM group has the highest tan δ due to the weakening of the elastic network. In the high-temperature region, elasticity dominates, and tan δ decreases. The LOM group has the lowest tan δ due to the strong rigidity of the network, exhibiting a "strong elasticity-weak viscosity" advantage.

[0111] like Figure 2 As shown in D, with increasing shear rate, the apparent viscosity of the minced meat gradually decreases, exhibiting significant shear-thinning behavior and displaying typical non-Newtonian fluid characteristics. In LOHM, lard and lycopene are first dissolved and homogenized into an emulsion before being blended with the meat into a paste. The high shear force during emulsification refines the oil droplets and evenly disperses them within the minced meat. The formation of the emulsion may result in a more uniform distribution of fat and water. The fine oil droplets and stable emulsion reduce internal molecular chain friction and flow resistance, exhibiting the lowest apparent viscosity.

[0112] 5. Effects on lipid oxidation (1) Test method The thiobarbituric acid reactive substances (TBARS) values ​​of three cell-cultured beef meatballs (M, LM, and LOHM) were determined according to the method described in "Grape seed extract as antioxidant in cooked, cold stored Turkeymeat" (DOI: 10.1016 / j.lwt.2005.02.003). These values ​​were used to measure the degree of lipid oxidation; higher values ​​indicated more severe lipid oxidation. The specific method was as follows: 5 g of sample was homogenized in 50 mL of 7.5% trichloroacetic acid (containing 0.1% EDTA) and then filtered using Whatman filter paper. 5 mL of the filtrate was then added to 5 mL of 0.02 mol / L thiobarbituric acid solution and reacted in boiling water at 100°C for 1 h. After cooling to room temperature, the absorbance was measured at 532 nm. The TBARS content was calculated using a standard curve of 1,1,3,3-tetraethoxypropane, and the results were expressed as mg MDA / kg in the sample.

[0113] (2) Test results like Figure 3 As shown, the TBARS value of the group with added lycopene and homogenized with lard before addition (LOHM) was significantly lower than that of the blank control group (M) without addition. However, the TBARS value of the group with direct addition of lycopene (LM) did not show a significant decrease. This may be because lycopene loses its antioxidant activity during the direct addition process, while lard can encapsulate lycopene and inhibit its own oxidation, thereby reducing the overall lipid oxidation degree of the meatballs. Among them, the TBARS value of the LOHM group was the lowest, and the antioxidant effect was the most prominent. The oil homogenization control group (OH group) had a relatively higher TBARS value than the LOHM group because it did not contain lycopene, and the lipid oxidation was more obvious.

[0114] 6. Effects on protein oxidation (1) Test method Carbonyl group determination: Carbonyl groups are a core marker of protein oxidative damage. The carbonyl content of three cell-cultured beef meatballs (M, LM, and LOHM) was determined according to the 2,4-dinitrophenylhydrazine (DNPH) method described in "Age-related changes in oxidized proteins." The specific method is as follows: 2 g samples were homogenized using 10 mL of 20 mmol / L phosphate buffer (pH 6.5). Then, 1 mL of 40% trichloroacetic acid was added to 200 μL of homogenate to concentrate the protein, and the mixture was centrifuged at 15000 ×g for 1 min. HCl containing 10 mmol / L DNPH was added to the sample group, and an equal volume of DNPH-free HCl was added to the control group (for background correction). The reaction tubes were then incubated at 30 °C for 1 h. 1 mL of 40% TCA was added to the liquid, and the mixture was centrifuged at 15000 ×g for 1 min at 4 °C to remove the supernatant. The precipitate was washed with ethyl acetate-ethanol solution (ethyl acetate:ethanol = 1:1, v / v) and centrifuged (15000 × g, 1 min, 4 °C), discarding the supernatant. These steps were repeated until the supernatant became colorless and transparent. The supernatant was discarded, and the precipitate was dissolved in 6 mol / L guanidine hydrochloride. The absorbance of the carbonyl solution was measured at 370 nm. Protein concentration was determined using a BCA microplate assay kit, and a 22000 cm⁻¹ plate was used. -1 Carbonyl content was calculated using the molar absorptivity of the carbonyl group. Results were expressed as nmol carbonyl groups / mg protein.

[0115] Determination of free thiol groups: The free thiol content of three cell-cultured beef meatballs (M, LM, LOHM) was determined according to the method described in "Physicochemical change and protein oxidation inporcine longissimus dorsi as influenced by different freeze-thaw cycles" (DOI:10.1016 / j.meatsci.2009.05.003). 1 g of sample was added to 10 mL of buffer (0.086 mol / L Tris, 0.09 mol / L glycine, 4 mmol / L EDTA, pH 8.0) and homogenized at 10000 r / min (pausing every 30 s for 30 s, repeated 3 times). The sample was then centrifuged at 10000 r / min for 15 min, and the supernatant was collected. Take 2 mL of supernatant, add 40 μL of Ellman's reagent (10 mM DTNB), and shake at room temperature in the dark for 1 h. Record the absorbance at 412 nm. Protein concentration is determined using a BCA microplate assay kit, and a 13600 cm⁻¹ plate is used.-1 The free thiol content was calculated using the molar absorptivity of the free thiol groups. The results are expressed as nmol free thiols / mg protein.

[0116] (2) Test results like Figure 4 As shown in Figure A, M has the highest carbonyl content. Due to the lack of antioxidant intervention, the protein is attacked by reactive oxygen species generated by lipid oxidation, resulting in severe oxidative damage. The protein is prone to oxidative deterioration, affecting its quality. In contrast, LM and LOHM, which have added lycopene, have lower carbonyl content than M, indicating that their protein oxidation is lower. LOHM performs best in terms of carbonyl content because its addition process (dissolution + homogenization) ensures that lycopene is evenly dispersed, thus fully exerting its antioxidant effect and inhibiting protein carbonylation.

[0117] Trends in free sulfhydryl content in cell-cultured meatballs under different lycopene addition methods Figure 4 As shown in Figure B, the free sulfhydryl content of M and LOHM is comparable, while that of LM is the lowest. Overall, the presence and dispersion of lycopene significantly affect the degree of protein oxidation. In M without added lycopene and LOHM with lycopene added via oil-soluble homogenization, the free sulfhydryl content is relatively high, indicating less protein oxidation; while in LM with direct mixing, this index decreases significantly, indicating a higher degree of protein oxidation.

[0118] Similar results were observed in other independent experiments when the amount of lycopene added to LM and LOHM was 0.075%wt, 0.100%wt, or 0.150%wt, respectively.

[0119] The results above indicate that the addition of lycopene can significantly improve the flavor, texture, stability, and antioxidant properties of cell-cultured beef meatballs.

[0120] Test Example 2: The effect of lycopene addition method on the digestibility of cell-cultured beef meatballs This embodiment evaluates the digestibility of M, LM and LOHM prepared in Examples 1-3 from the perspectives of degree of hydrolysis, surface hydrophobicity, particle size, DPPH free radical scavenging and ABTS free radical scavenging through in vitro simulated digestion experiments.

[0121] 1. Testing Method (1) In vitro simulated digestion Electrolyte stock solutions for gastrointestinal fluids were prepared in accordance with the standard "A standardized static in vitro digestion method suitable for food - an international consensus" (DOI: 10.1039 / c3fo60702j), and the specific formula is shown in Table 6.

[0122] Table 6 Electrolyte Reserves for Gastrointestinal Fluids

[0123] Because protein digestion primarily occurs in the stomach and small intestine, this test did not include the oral digestion phase. Instead, in vitro digestion simulations of the stomach and intestines were performed according to the standardized INFOGEST protocol outlined in "INFOGEST static in vitro simulation of gastrointestinal food digestion" (DOI:10.1038 / s41596-018-0119-1). The specific methods are as follows: During the gastric digestion stage, 5 mL of simulated gastric juice (SGF) containing porcine pepsin (3000 U / mg) was added to 5 g of sample, resulting in a final porcine pepsin activity of 25,000 U / mL. The pH was adjusted to 3.0 with 6 mol / L HCl, and the sample was incubated at 37°C with shaking for 2 h. The gastric digested sample (denoted as G) was then collected.

[0124] During the intestinal digestion stage, 20 mL of simulated intestinal fluid (SIF) containing porcine trypsin (250 U / mg) was added to the sample after gastric digestion, resulting in a final trypsin activity of 800 U / mL. The pH was adjusted to 7.0 with 1 mol / L NaOH, and the mixture was thoroughly mixed at 37°C for 2 h. The intestinal digested sample (denoted as G+I) was then collected.

[0125] After intestinal digestion, the sample was heated at 95°C for 5 min to stop the enzymatic reaction, centrifuged (10000×g, 15 min, 4°C), and the supernatant and precipitate were collected and stored at -80°C for later use.

[0126] (2) Determination of degree of hydrolysis Degree of hydrolysis (DH) is used to assess the digestibility of proteins. The degree of hydrolysis after trypsin treatment is higher than that after pepsin treatment alone. The study "Structural modification and digestibility change of β-lactoglobulin modified by methylglyoxal with the simulated reheating of dairy products" (DOI: 10.1016 / j.foodchem.2019.03.021) found that the DH of pepsin-treated wheat bran was 20%, while the DH increased to approximately 50%–60% after treatment with both pepsin and trypsin. This indicates that pepsin plays an important role in breaking down peptide bonds in large protein components in the digestive system, followed by further degradation by trypsin, reducing the size of small protein components and long-chain peptides to small peptides and amino acids.

[0127] Following the method described in "Digestion of protein and protein gels in simulated gastric environment" (DOI: 10.1016 / j.lwt.2015.03.087), the degree of protein hydrolysis in three types of cell-cultured beef meatballs during gastric and intestinal digestion was detected using the OPA method. The specific method is as follows: Preparation of OPA reagent: Add 80 mg OPA to 2 mL of anhydrous ethanol and dissolve in the dark; dissolve 1.9068 g sodium tetraborate, 0.1 g SDS, and 88 mg DTT in water; transfer to a 100 mL brown volumetric flask and dilute to volume with distilled water.

[0128] Construction of the standard curve: Dissolve 10 mg L-serine standard in 100 mL of deionized water to prepare a 0.1 mg / mL standard solution. Pipette 100, 200, 300, and 400 μL of the serine standard solution into centrifuge tubes, respectively, and make up to 400 μL with deionized water. Add 3 mL of OPA reagent, react for 2 min, and then measure the absorbance at 340 nm.

[0129] (3) Surface hydrophobicity Following the method described in "The effect of meat processing methods on changes in disulfidebonding and alteration of protein structures: impact on protein digestion products" (DOI: 10.1039 / c8ra02310g), the surface hydrophobicity of proteins in meatballs (M, LM, and LOHM) was measured using 8-aniline-1-naphthalenesulfonic acid (ANS). The total protein concentrations of each meatball were diluted to 0.05 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.375 mg / mL, and 0.5 mg / mL, respectively. 4 mL of each protein dilution was added to 20 μL of ANS, and the mixture was incubated in the dark for 10 min. Excitation wavelength was 374 nm, and emission wavelength was 480 nm. Phosphate buffer solution was used as a blank control. Surface hydrophobicity was expressed as the slope of the fluorescence intensity versus protein concentration curve.

[0130] (4) Particle size determination Following the method described in "The Effects of Dynamic Pressure Swing Marinating on the Eating Quality and Protein Digestibility of Roast Duck", the total protein samples of each beef ball were centrifuged (10000 × g, 15 min, 4℃). The supernatant was then collected, and the particle size was determined using a Malvern Mastersizer 3500 laser particle size analyzer with the following parameters: non-spherical particle type and density of 1 g / cm³. 3 The dispersant is deionized water.

[0131] (5) Determination of DPPH free radical scavenging rate Samples (G, G+I) from each stage of the in vitro simulated digestion process were diluted 20-fold with deionized water and used as test samples. Their DPPH free radical scavenging ability was determined using the following method: 100 μL of the test sample was added to an equal volume of 200 μM DPPH ethanol solution and mixed thoroughly to form the sample group; the blank group used anhydrous ethanol without DPPH instead of the DPPH ethanol solution; and the correction group used an equal volume of pure water instead of the test sample. Each group was reacted in the dark for 30 min, and the absorbance was measured at 517 nm.

[0132] Based on the test results, the DPPH free radical scavenging rate was calculated using the following formula: ; Among them, A 样品 A represents the absorbance value at 517 nm for the sample group. 空白 A represents the absorbance value at 517 nm for the blank group. 矫正This indicates the absorbance value at 517 nm for the corrected group.

[0133] (6) Determination of ABTS free radical scavenging rate Samples (G, G+I) from each stage of the in vitro simulated digestion process were diluted 5 times with deionized water and used as test samples. ABTS + The free radical scavenging ability was assessed using the following method: An equal volume of ABTS ethanol solution (7 mM) and 4.9 mM potassium persulfate aqueous solution were mixed and reacted at room temperature (25°C) in the dark for 12–16 h to obtain ABTS. + Stock solution. Before assay, prepare ABTS. + The stock solution was diluted to an absorbance of 0.7 ± 0.02 at 734 nm. + Prepare the working solution fresh before use. Take 10 μL of the test sample and add 200 μL of ABTS. + The working solution was thoroughly mixed and used as the sample group; the blank group was prepared by replacing the test sample with an equal volume of pure water. Each group was reacted in the dark for 6 minutes, and the absorbance was measured at 734 nm.

[0134] Based on the test results, ABTS was calculated. + The free radical scavenging rate is calculated using the following formula: ; Among them, A 样品 A represents the absorbance value at 734 nm for the sample group. 空白 This represents the absorbance value at 734 nm for the blank group.

[0135] 2. Test Results like Figure 5 As shown in Figure A, after gastric digestion, the hydrolysis degree (DH) of M and LM was comparable, at 4.5% and 4.4%, respectively, while the DH of LOHM was significantly higher (4.8%). After intestinal digestion, the DH of M, LM, and LOHM all increased, at 35.2%, 32.0%, and 27.5%, respectively. This indicates a trend of increasing DH as digestion progresses. During gastric digestion, the degree of hydrolysis was generally low across all groups with small differences between groups, but significantly increased after entering the gastrointestinal digestion stage. Regarding the method of addition, the degree of hydrolysis was relatively higher in the blank control group (M) during the gastrointestinal stage, while the increase in the degree of hydrolysis in the lycopene oil-soluble addition groups (LM, LOHM) was relatively smaller. This suggests that the addition of lycopene first protects meat proteins during gastric digestion, preventing excessive hydrolysis, and then promotes the hydrolysis of meat proteins during intestinal digestion, thereby promoting digestion and absorption.

[0136] like Figure 5As shown in Figure B, before digestion, the surface hydrophobicity of M and LM was comparable, at 115.5% and 120.0%, respectively, while the surface hydrophobicity of LOHM was significantly lower (68.5%). After gastric digestion, the surface hydrophobicity of M, LM, and LOHM all decreased, at 22.5%, 27.5%, and 30.8%, respectively. After intestinal digestion, the surface hydrophobicity of M, LM, and LOHM all decreased further, at 10.8%, 10.7%, and 9.8%, respectively, indicating that group M was not protected by lycopene. In the LM group, the lycopene was not homogenized, which caused the protein to undergo slight denaturation during early processing (grinding, chopping, etc.). This exposed hydrophobic groups from the molecular interior to the surface (high surface hydrophobicity), leading to protein aggregation and hindering enzymatic hydrolysis. In the LOHM group, the lycopene's oil-soluble dispersion stabilized the protein, or the homogenized and refined fats interacted more tightly with the protein interface, inhibiting the exposure of hydrophobic groups (low surface hydrophobicity). This resulted in a relatively higher degree of hydrolysis in the LOHM group during gastric digestion, promoting pepsin activity. However, the LOHM group exhibited the highest surface hydrophobicity during gastric digestion. This may have been due to the added lycopene promoting pepsin activity, leading to a stronger surface hydrophobicity in meat proteins (e.g., a large number of exposed hydrophobic amino acid residues), forming tight hydrophobic aggregates (e.g., protein precipitation, lipid clusters). This made it difficult for enzymes to access the internal hydrolysis sites, thus reducing the degree of hydrolysis.

[0137] Table 7. Effects of lycopene addition method on particle size of beef meatballs cultured after simulated digestion in vitro.

[0138] Note: D 3,2 D is the diameter with average surface area. 4,3 The volume-average diameter is represented by Dx(10), Dx(50), and Dx(90), which indicate that 10%, 50%, and 90% of the particles are smaller than this diameter, respectively; different lowercase letters indicate significant differences ( P <0.05).

[0139] As shown in Table 7, during the gastric digestion stage, LOHM promotes particle aggregation due to homogenization, thus reducing D... 3,2 D 4,3 The cumulative particle size increased; M and LM showed no effective aggregation and had even smaller particle sizes. During intestinal digestion, the M group received no intervention, and the particles were fully broken down by enzymatic hydrolysis, resulting in even smaller particle sizes.

[0140] In other independent experiments, similar results were observed when the lycopene content in LM and LOHM was 0.075% wt, 0.100% wt, or 0.150% wt. However, the beef meatballs prepared in Example 4, despite the addition of lycopene, showed significantly lower hydrolysis compared to LOHM, making them more difficult to digest.

[0141] The above results indicate that adding lycopene can significantly improve the digestibility of cell-cultured beef meatballs, increase nutrient supply efficiency, and reduce the digestive burden on consumers.

[0142] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for improving the quality and digestibility of meat products, characterized in that, The method includes adding cell-cultured meat and lycopene in a mass ratio of (90-98):1 to the raw materials of meat products.

2. The method according to claim 1, characterized in that, Add 4%wt to 5%wt of cell-cultured meat and / or 0.04%wt to 0.06%wt of lycopene to the raw materials of the meat product.

3. The method according to claim 1 or 2, characterized in that, The raw materials for the meat products are selected from any one or two of animal meat and animal fat; The mass ratio of the cell-cultured meat to the animal meat is 3:(40-60), and / or, The mass ratio of lycopene to animal oil is 1:(110-130).

4. The method according to claim 3, characterized in that, The mass ratio of the animal meat to the animal oil is (35-45):

3.

5. The method according to claim 3 or 4, characterized in that, The method includes: first mixing the lycopene with the animal oil, then mixing the resulting mixture with other components in the raw materials to obtain minced meat.

6. The method according to any one of claims 1 to 5, characterized in that, The cell-cultured meat includes cell-cultured meat of poultry origin, preferably cell-cultured meat of pigeon origin.

7. The method according to claim 6, characterized in that, The cell-cultured meat includes: muscle tissue and adipose tissue obtained by differentiation and culture of muscle satellite cells; Preferably, the mass ratio of the muscle tissue to the adipose tissue is (8-12):

1.

8. The method according to any one of claims 1 to 7, characterized in that, Improving the quality of meat products includes any of the following: (1) Improve the sensory evaluation of meat products; (2) Improve the texture of meat products; (3) Inhibit the oxidation of protein and / or fat in meat products.

9. The method according to any one of claims 1 to 7, characterized in that, Improving the digestibility of meat products includes any of the following: (1) Increase the degree of hydrolysis of meat products; (2) Improve the surface hydrophobicity of meat products; (3) Reduce the particle size of meat products.

10. A meat product, characterized in that, Prepared by the method according to any one of claims 1 to 9; Preferably, the meat product is a meatball, meat patty, sausage, or chunk of meat.

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