Artificial meat seasoning method based on palmitoleic acid or natural or genetic engineering biological products containing palmitoleic acid

Abstract

The invention discloses artificial meat and a preparation method thereof. The artificial meat contains palmitoleic acid with the mass percent of 0.01% or above. The invention also provides an artificial meat flavoring method based on palmitoleic acid or natural or genetic engineering biological products containing palmitoleic acid. According to the method, the health benefits of palmitoleic acid are fused, the natural raw materials rich in palmitoleic acid are combined with traditional flavoring agents to be applied to the artificial meat to achieve the flavoring effect comparable with hemoglobin, and satisfaction and taste enjoyment similar to those of real meat can be provided. The vegetable protein meat is prepared by introducing palmitoleic acid, so that not only is a low-fat and cholesterol-free meat substitute provided, but also the taste and nutritional value of the product are improved through continuous innovations of a pretreatment mode, selection of auxiliary materials and the like, so as to attract more consumers seeking healthy lifestyles.

Description

Technical Field

[0001] The present invention belongs to the technical field of food, specifically to the technical field related to the flavoring technology of artificial meat products, and particularly relates to a method for flavoring artificial meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid. Background Art

[0002] As of December 2023, the global plant-based meat market is experiencing rapid growth. This growth is mainly driven by the following factors: Consumers are increasingly concerned about healthy eating, and plant-based meat is favored for its low-fat and low-cholesterol characteristics. Concerns about climate change and environmental sustainability have prompted people to seek food choices with less impact on the planet, and plant-based meat, as an alternative, can reduce dependence on the livestock industry. The rise of the animal rights movement has led some consumers to switch to plant-based meat products to avoid consuming meat from animal slaughter. Advancements in food technology, especially breakthroughs in flavor and texture imitation, have made plant-based meat products increasingly similar to the real meat experience. Food technology companies have received a large amount of investment, which has promoted product R & D and market expansion. Governments in some countries have started to support the R & D and commercialization of alternative proteins such as plant-based meat in order to address climate change and improve food safety.

[0003] Nonetheless, the plant-based meat market also faces challenges, including consumer acceptance of genetically engineered foods, differences in acceptance of plant-based meat across different cultures, and competition from the traditional meat industry. Overall, the plant-based meat market is expected to continue to expand, especially in some key markets in North America, Europe, and Asia. With the continuous improvement of product variety and quality, as well as the gradual change of consumer habits, plant-based meat has the potential to become an important part of global food consumption. With the popularity of plant-based protein meat products, various brands in the market, such as Beyond Meat, Quorn, and Omnipork, are constantly exploring ways to enhance the flavor of their products to make them closer to the taste and texture of traditional meat. In this process, the use of umami agents becomes particularly important. In addition to Impossible Foods' unique use of hemoglobin, most other plant-based meat brands usually add various umami agents to enhance the taste of their products. These umami agents include, but are not limited to, traditional seasonings such as chicken essence and mushroom essence, which enhance the umami of food by extracting the flavors of natural ingredients. Chicken essence is a concentrated flavor extract from chicken, although it is necessary to ensure that its source is non-animal when used in plant-based protein meat products. Mushroom essence is the flavor extracted from mushrooms, which gives plant-based meat an earthy and deep flavor similar to natural meat, but there are still differences. In addition, synthetic umami agents such as Disodium 5'-ribonucleotides are commonly used in the food industry to replicate the umami of meat. Their use in plant-based protein meat products can mimic the complex flavors of meat and provide a more profound umami experience. Other synthetic umami agents may also include monosodium glutamate (MSG) and yeast extract, which are flavor enhancers that can bring a more intense taste to food without adding excessive calories. Impossible Foods is a well-known plant-based meat company. Its core product, the Impossible Burger, mimics the taste and texture of traditional burger meat in a unique way, especially because it uses a special ingredient - synthetic heme, which is one of the main innovations of its products. Heme is an iron-containing bioactive substance that exists in the cells of all living organisms, especially in higher concentrations in the blood and muscle tissues of animals. It is one of the key components contributing to the unique flavor of meat. In animal meat, heme contributes to the "bloody" taste and red appearance of the meat. Impossible Foods uses yeast through a fermentation process to produce plant-based heme, which is called soy leghemoglobin and has a similar function to animal heme but is derived from plants. Impossible Foods first applied genetic engineering to yeast to enable it to produce soy leghemoglobin.Then, through a fermentation process similar to that in beer or bread making, yeast cells produce leghemoglobin. This process allows Impossible Foods to mass-produce this compound without using animal ingredients, providing a plant-based meat alternative with meaty flavor and texture for vegetarians and those looking to reduce meat consumption. The use of this synthetic hemoglobin is one of the main features that differentiates Impossible Foods' products from other plant-based meat products. It not only enhances the flavor of the product, making the plant-based meat closer to the taste of real meat, but also increases the similarity in color and texture, thus achieving significant success and recognition in the market. This technological innovation by Impossible Foods has also sparked extensive discussions on food technology and sustainable food production, marking a major step forward in the imitation of real meat flavor and texture by plant-based meat products. However, this innovation has also drawn the attention of regulatory agencies because it involves genetic engineering and new food ingredients. In the United States, leghemoglobin has been recognized by the U.S. Food and Drug Administration (FDA) as safe for consumption. But in other countries, the safety of the application of this soy leghemoglobin in food still needs to be reviewed and approved by relevant departments.

[0004] The gap in taste from real meat is the biggest obstacle to the development of artificial meat and an urgent problem to be solved in its current development. In artificial meat products, the choice of flavorings is crucial and currently includes natural umami substances, artificial synthetic umami substances, and artificial hemoglobin. Natural umami substances, such as amino acids and nucleotides, although generally considered safe and able to provide rich flavors, have high costs and flavor consistency may vary with seasons and raw material sources. In contrast, artificial synthetic umami substances like monosodium glutamate are cost-effective and ensure the stability of the product flavor, but may face consumer health concerns and issues of simplified flavors. Artificial hemoglobin, especially soy leghemoglobin, can mimic the taste and color of real meat, representing innovation and sustainability in food technology, although it may face high production costs and challenges in consumer acceptance; moreover, currently, the patents and technologies for actually usable soy leghemoglobin in production are monopolized by the U.S. company Impossible Foods. Therefore, in the specific product production process, other producers need to weigh multiple factors such as costs, consumer preferences, product positioning, compliance with local regulations, and market dynamics when choosing flavorings, and currently, there are not many flavorings available for artificial meat, which also becomes the main factor restricting the development of the artificial meat industry.

[0005] The flavor of meat is the result of the combined action of various compounds, including fatty acids, amino acids, sugars, nucleotides, etc. During the heating and cooking process, these components generate various flavor substances through chemical reactions such as the Maillard reaction and lipid oxidation. The main components of meat flavor substances include Maillard reaction products, fatty acid oxidation products, sulfur-containing compounds in muscle tissue, and sulfur-containing compounds in muscle tissue. Maillard reaction products are produced by the Maillard reaction, which mainly occurs between amino acids and reducing sugars, generating a series of complex flavor compounds, such as heterocyclic compounds like pyrrole, thiophene, pyridine, and caramel compounds. Chemical reactions such as lipid oxidation generate various flavor substances, which are various volatile molecules such as aldehydes, ketones, and acids produced by the oxidation of fatty acids during cooking. These molecules have a great impact on the flavor of meat. The production of sulfur-containing compounds in muscle tissue is due to the decomposition of sulfur-containing amino acids such as cysteine at high temperatures to produce thiols, thioethers, etc. These compounds are crucial for the formation of meat flavor. Sulfur-containing compounds in muscle tissue refer to inosinic acid, guanylic acid, etc., which are umami compounds produced by decomposition during cooking.

[0006] Advantages and disadvantages of current commonly used meat flavor enhancers: Amino acid-based (such as sodium glutamate (monosodium glutamate), disodium inosinate) have the advantage of having a strong umami flavor, being able to significantly enhance the flavor of food, having a relatively low cost, and being easy to produce on a large scale; however, their disadvantage is that excessive use may lead to the monosodium glutamate syndrome, and some consumers have health concerns about monosodium glutamate. Plant protein hydrolysates (such as soy protein hydrolysates) have the advantage of being able to provide complex flavor characteristics, including umami and meaty flavors. They are usually friendly to vegetarians. However, their disadvantage is that the flavor may not be as good as animal-derived extracts. Some people may have allergic reactions to soybeans or other raw materials. Yeast extracts have the advantage of being able to provide a natural umami flavor and taste and can be used as a substitute for monosodium glutamate. However, their disadvantage is that the cost is relatively high, and the flavor intensity may not be as good as chemically synthesized flavoring agents. Synthetic meat flavorings (such as smoked flavorings) have the advantage of having a strong specific meat flavor, such as smoked flavor and grilled meat flavor, and can maintain flavor consistency. However, their disadvantage is that the taste is not natural enough, contains a large amount of artificial additives (including a large amount of sulfur-containing compounds), poses a health risk, and causes concerns. There is an urgent need to develop a flavoring agent that is superior to the above-mentioned conventional meat flavoring agents and flavor enhancers, can rival hemoglobin, is green, pollution-free, beneficial to health, and widely applicable to artificial meat products, and its application.

[0007] Although plant-based meat products on the market have achieved certain success, there has been no attempt to incorporate palmitoleic acid, a component with significant health benefits, into plant-based meat products. OMEGA-7 fatty acids, especially palmitoleic acid, have shown potential benefits in promoting skin health, improving metabolism, maintaining cardiovascular health, and regulating blood sugar levels in clinical studies. However, the prices of current palmitoleic acid products are generally high. Although there are many natural products rich in palmitoleic acid, there are few reports on their flavoring effects in plant-based artificial meat. Therefore, how to add palmitoleic acid and set the specific addition amount, etc., are all unknown situations. Summary of the Invention

[0008] Object of the Invention: The technical problem to be solved by the present invention is to provide an artificial meat that can produce a taste similar to or the same as that of meat products, and even enhance the complexity and layering of meat flavors, achieving a flavoring effect similar to that of adding hemoglobin.

[0009] Another technical problem to be solved by the present invention is to provide a preparation method of artificial meat.

[0010] Finally, the technical problem to be solved by the present invention is to provide a flavoring method for artificial meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid.

[0011] Technical Solution: To solve the above technical problems, the present invention provides an artificial meat, which contains palmitoleic acid with a mass percentage of more than 0.01%. Preferably, the mass percentage of palmitoleic acid is 0.01 - 0.07%.

[0012] Among them, the palmitoleic acid in the artificial meat is derived from natural or genetically engineered biological products rich in palmitoleic acid. Preferably, the natural products rich in palmitoleic acid include one or more of sea buckthorn juice, sea buckthorn oil, macadamia nut oil, or avocado oil. Preferably, the genetically engineered biological products rich in palmitoleic acid include products containing palmitoleic acid produced by engineered bacteria or plants optimized and constructed through gene mutagenesis, gene editing, or transgenic technology.

[0013] The present invention adds a certain amount of palmitoleic acid or vegetable oil or microbial fermentation product containing palmitoleic acid to produce flavor substances through thermal decomposition for flavoring and simultaneously provide health care effects.

[0014] Among them, the artificial meat further includes main ingredients and auxiliary ingredients. Preferably, the main ingredients include a base composed of a plant protein matrix, and the auxiliary ingredients include natural products and natural flavors of other plants or yeasts.

[0015] Among them, the plant protein matrix includes one or several of soy protein, wheat protein, or potato protein. Preferably, the excipients include one or more of resistant dextrin, vital gluten, beet powder, mushroom essence, sunflower oil, coconut oil, or yeast extract.

[0016] Among them, the artificial meat of the present invention includes at least yeast extract containing palmitoleic acid and / or natural or genetically modified plant products; these products are obtained through specific biological processes to produce a flavor effect similar to the thermal decomposition of palmitoleic acid; the yeast extract containing palmitoleic acid can replace 36 - 40 parts of yeast extract described in the above formula, and the genetically modified products can include vegetable oil and potato protein (extract) and their substitutes, etc., and activate their flavor substances through the above heat treatment process. (Note: Commercially available yeast extracts usually do not contain or only contain extremely trace amounts of palmitoleic acid.)

[0017] Among them, the artificial meat is made from the following components by weight: 0.1 - 0.7 parts of palmitoleic acid, 200 - 210 parts of soy protein, 20 - 25 parts of potato protein, 8 - 10 parts of resistant dextrin, 100 - 130 parts of vital gluten, 16 - 18 parts of beet powder, 36 - 40 parts of yeast extract, 6 - 7 parts of mushroom essence, 24 - 25 parts of rapeseed oil, 18 - 20 parts of coconut oil, and 0.2 - 0.25 parts of salt.

[0018] The present invention also includes a method for preparing the artificial meat described above, comprising the following steps:

[0019] 1) Mixing some raw materials and / or some pre - treated raw materials to obtain a pre - mixture;

[0020] 2) Adding the pre - mixture to other raw materials and mixing evenly to obtain a plant - based meat mixture, and the other raw materials include natural or genetically engineered biological products rich in palmitoleic acid;

[0021] Preferably, the preparation method further includes putting the plant - based meat mixture obtained in step 2) into a mold and compacting it with appropriate pressure.

[0022] Among them, the preparation method specifically includes the following steps:

[0023] 1) Accurately weigh the following raw materials: soy protein, potato protein, resistant dextrin or resistant starch, vital gluten, beet powder, palmitoleic acid or natural products rich in palmitoleic acid, yeast extract, mushroom essence, sunflower oil, organic coconut oil, and salt;

[0024] 2) Pretreatment of soy protein: Soak soy protein in hot water, keep the temperature controlled at 95 - 100 °C, and ensure sufficient water absorption. After soaking, drain it using a sieve and cool it at room temperature;

[0025] 3) Mixing and homogenizing the ingredients: Mix the soy protein and wheat gluten after the above pretreatment with vegetable oils (refined rapeseed oil and coconut oil) to ensure the mixing of wheat gluten and soy protein and the thorough mixing of the oils, obtaining a pre-mixture; place the pre-mixture together with other raw materials in a mixer; mix at a controlled speed and for a certain time until a homogeneous and consistent plant-based meat mixture is obtained;

[0026] Preferably, the rotation speed is 120 - 150 rpm and the time is 10 - 15 min.

[0027] The present invention also includes a method for flavoring plant-based meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid. The method for flavoring plant-based meat is to add palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid to the main plant protein matrix and other auxiliary materials for heat treatment.

[0028] Among them, the heating temperature is 150 °C and the heating time is 20 minutes.

[0029] The method for flavoring plant-based meat according to the present invention specifically includes the following steps:

[0030] a. Provide a plant protein matrix;

[0031] b. Add palmitoleic acid or a vegetable oil or microbial fermentation product containing palmitoleic acid to the matrix;

[0032] c. Subject the mixture after the above flavoring technical treatment to heat treatment to generate flavor substances.

[0033] Among them, the heat treatment process according to the present invention involves heating the mixture to above 90 °C, and this temperature range is sufficient to cause the decomposition of palmitoleic acid contained therein or to react with other substances to generate a mixed aroma containing the above flavor substances.

[0034] Through experiments, the present invention found that a series of volatile substances are produced after palmitoleic acid decomposes under heat. After adding palmitoleic acid or a vegetable oil containing palmitoleic acid to the plant-based meat formula of the present invention, cooking will produce a pleasant aroma similar to that of meat. The flavoring technology of the present invention centered on palmitoleic acid and biological products rich in palmitoleic acid includes an optimized thermal decomposition process of palmitoleic acid, thereby generating flavor substances such as peach aldehyde / myristaldehyde, 2-methoxy-4-vinylphenol, octanal, hexanal, 1-octen-3-ol, 2-n-amylfuran, nonanal, maltol, octanoic acid, decanal, (E,E)-2,4-nonadienal, nonanoic acid, (E,E)-2,4-decadienal, (E,Z)-2,4-decadienal, (E)-4-decenal, (E)-2-tetradecenol, (E)-2-undecenal, etc. These substances are also important factors for hemoglobin to decompose under heat to form fragrance in the plant-based meat mixture.

[0035] Specifically, these volatile compounds have the following effects: Peach aldehyde / myristaldehyde (2(3H)-Furanone,5-heptyldihydro-): fruity and creamy odor. 2-Methoxy-4-vinylphenol: has the odors of apple, peanut and curry. Octanal: has fruity and citrus scents. Hexanal: produces a grassy aroma. 1-Octen-3-ol: has a mushroom aroma. 2-N-Amylfuran: has a bean and fruity smell. Nonanal: produces citrus and oily odors. Maltol: sweet, caramel flavor. Octanoic acid: fruity sour taste. Decanal: sweet citrus peel, citrusy and fatty floral scent. (E,E)-2,4-Nonadienal: has a fresh cucumber-like scent. Nonanoic acid: cheesy aroma. (E,E)-2,4-Decadienal: has the flavors of roasted chicken and grease. (E,Z)-2,4-Decadienal: fried flavor. (E)-4-Decenal: orange and chicken aromas. (E)-Tetradec-2-enal: lemon and waxy odors. (E)-2-Undecenal: fruity aroma and waxy odor.

[0036] These compounds disclosed in the present invention are all the main components of the aroma substances after meat heating. By adding palmitoleic acid and oils rich in palmitoleic acid to products such as plant-based meat and subjecting them to heat treatment to decompose them into these substances, these substances provide the complexity and diversity of meat flavor for plant-based meat products. By precisely controlling the decomposition conditions of palmitoleic acid, the yield of these volatile compounds can be optimized, and thus, without using animal ingredients, a flavor experience similar to that of traditional meat products can be created. These volatile compounds can also combine with other ingredients such as proteins, fats and carbohydrates, and through reactions such as the Maillard reaction and lipid oxidation during the cooking process, further enhance the complexity and layering of meat flavor. Therefore, by simulating the chemical reactions that occur during meat cooking, plant-based meat products can provide a healthy and sustainable meat alternative without sacrificing flavor.

[0037] Beneficial effects: Compared with the prior art, the present invention has the following advantages: The present invention discloses a brand-new artificial meat product, specifically a plant-based protein meat product, which not only incorporates the health benefits of palmitoleic acid, such as health care effects like regulating metabolism and endocrine, but also achieves a flavor effect comparable to hemoglobin by applying natural raw materials rich in palmitoleic acid combined with traditional flavoring agents in the artificial meat, and can provide a sense of satisfaction and gustatory enjoyment similar to that of real meat. The innovative flavor enhancement strategy of the present invention is expected to meet the market demand for healthy, natural, and delicious plant-based protein meat products, while providing a more sustainable and environmentally friendly food option for consumers and artificial meat manufacturers. The present invention prepares plant-based protein meat by introducing palmitoleic acid, which not only provides a low-fat and cholesterol-free meat substitute, but also continuously innovates through pretreatment methods, selection of auxiliary materials, etc. to improve the taste and nutritional value of the product, so as to attract more consumers seeking a healthy lifestyle. Description of the Drawings

[0038] Figure 1 Principal component analysis (PCA) diagram of the flavors of the comparative example and the examples measured by an electronic nose;

[0039] Figure 2 Radar diagram of the electronic nose;

[0040] Figure 3 Release amount of specific fragrance volatile substances;

[0041] Figure 4 Ratio of the volatilization amount of different fragrance substances in different treatments to the volatilization amount in Comparative Example 1 (Note: * indicates that this substance has been detected in meat fragrance volatiles.)

[0042] Figure 5 Analysis of the intestinal flora diversity of different samples in in vitro fermentation experiments (Note: The Y-axis in the figure is the Shannon index, and the larger the coefficient, the higher the alpha diversity of the microorganisms in the sample.)

[0043] Figure 6 Relative abundances of different sample taxa at the phylum level in in vitro fermentation experiments of intestinal flora (Note: UPGMA (unweighted pair group method with arithmetic mean) clustering analysis was performed using a weighted UniFrac distance matrix (beta diversity, i.e., diversity between communities).)

[0044] Figure 7 Functional prediction based on the differences in intestinal flora of samples. Detailed Embodiments

[0045] The present invention will be further described below in conjunction with the drawings and embodiments.

[0046] Example 1 Preparation of artificial meat based on natural biological products containing palmitoleic acid

[0047] 1) Ingredient weighing steps:

[0048] Accurately weigh the following ingredients: 200 g non-GMO soy textured protein (Jinan Yuxin Biotechnology Co., Ltd., model: wet-milled small particles), 20 g high-quality potato protein (Jianpeptide Biotechnology (Shanxi) Co., Ltd., item number: JT-TDDBF), 8 g high-purity resistant dextrin / resistant starch (Anhui Qianshun Biotechnology Co., Ltd., item number: Q / CBL0008S); 100 g selected gluten powder (Henan Changjian Fine Chemical Co., Ltd., item number 3655244), natural beet powder (Shaanxi Guanchen Biotechnology Co., Ltd., item number: DSGSER) 16g, high-quality yeast extract 36g, pure mushroom essence 6g (manufacturer: Taitai Le); refined rapeseed oil 24ml (manufacturer: Jinlongyu), organic coconut oil 20ml (Guangdong Aimu Technology Co., Ltd., item number: MELAO-W027), freshly squeezed sea buckthorn juice 100ml (Note: frozen sea buckthorn fresh fruit produced in Northeast China, melted and pitted and homogenized, the fat content is about 2.0%, and the palmitic acid content is about 0.7%) and 2g fine-grained edible salt.

[0049] 2) Non-GMO soy protein hydrothermal treatment steps:

[0050] Soak 200 g of non-genetically modified soy protein in about 1.5 L of hot water, keep the temperature at 95-100° C., and last for 20 minutes to ensure sufficient water absorption; after soaking, drain with a sieve and cool to 25° C. at room temperature; then, put the non-genetically modified soy protein in a steam environment, maintain the temperature at 100° C., steam for 30 minutes, then drain and cool to room temperature (obtaining about 570 g of water-containing soy protein).

[0051] 3) Ingredient mixing and homogenization step:

[0052] Mix gluten, refined rapeseed oil and coconut oil in advance to ensure that the gluten can be evenly dispersed in the oil to obtain a premix; place the premix together with other raw materials (including the non-genetically modified soy protein treated in step 2) in a mixer; mix at a controlled speed and time until a homogeneous and consistent plant meat mixture is obtained, and the final product weighs 1KG, of which palmitic acid accounts for 0.07%.

[0053] 4) Molding process:

[0054] The evenly mixed plant meat mixture is placed into a pre-designed mold and compacted with appropriate pressure to form the desired product shape, namely artificial meat.

[0055] Example 2 Preparation of Cultured Meat Based on Natural Biological Products Containing Palmitoleic Acid

[0056] The preparation method of the embodiment of the present invention is the same as that of Example 1, except that 100 ml of sea buckthorn juice in Example 1 is replaced with 2 ml of sea buckthorn oil (the oil content of sea buckthorn juice is about 2.0%, and the palmitoleic acid content of sea buckthorn oil is about 35%), and the others remain unchanged. Among them, the content of palmitoleic acid in the cultured meat is about 0.7‰.

[0057] Example 3

[0058] The preparation method of the embodiment of the present invention is the same as that of Example 1, except that 100 ml of sea buckthorn juice in Example 1 is replaced with 0.7 g of palmitoleic acid (dissolved in 2 ml of salad oil), and the others remain unchanged. Among them, the content of palmitoleic acid in the cultured meat is about 0.7‰.

[0059] Example 4

[0060] The preparation method of the embodiment of the present invention is the same as that of Example 1, except that 100 ml of sea buckthorn juice in Example 1 is replaced with 2 ml of macadamia nut oil (the palmitoleic acid content is about 14.5%), and the others remain unchanged. Note: The content of palmitoleic acid in the cultured meat is about 0.29‰.

[0061] Example 5

[0062] The preparation method of the embodiment of the present invention is the same as that of Example 1, except that 100 ml of sea buckthorn juice in Example 1 is replaced with 2 ml of avocado oil (the palmitoleic acid content is about 5.5%), and the others remain unchanged. Note: The content of palmitoleic acid in the cultured meat is about 0.11‰.

[0063] Example 6

[0064] The preparation method of the embodiment of the present invention is the same as that of Example 1, except that 100 ml of sea buckthorn juice in Example 1 is replaced with 2 ml of olive oil, and the others remain unchanged. Note: Palmitoleic acid was not detected.

[0065] Example 7 Flavoring Method of Cultured Meat Based on Examples 1-5

[0066] The cultured meat of Examples 1-5 is further heat-treated. The uniformly mixed plant meat mixture is put into a pre-designed mold and compacted with appropriate pressure to form the desired product shape; then the product in the mold is transferred to a tray; in a steamer, the temperature is set and maintained at 150 °C, and the product is heat-treated for 20 minutes to achieve ideal texture and flavor characteristics.

[0067] Comparative Example 1

[0068] Replace 100 ml of sea buckthorn juice in Example 1 above with 2 ml of salad oil (the same as in Example 3), and keep the others unchanged. (Palmitoleic acid was not detected in the salad oil.)

[0069] Comparative Example 2

[0070] Replace 100 ml of sea buckthorn juice in Example 1 above with 2 ml of 50% bovine hemoglobin (Xi'an Yatu Biotechnology Co., Ltd., product number: YT-XHDBF230719) solution (1 g of bovine hemoglobin was dissolved in 2 ml of 0.1 mol / L NaOH solution), and keep the others unchanged. Note: The hemoglobin content is about 1‰.

[0071] Comparative Example 3

[0072] Commercially available beef processed into minced meat, and processed according to the heating treatment method of Example 7.

[0073] Comparative Example 4

[0074] Commercially available pork processed into minced meat, and processed according to the heating treatment method of Example 7.

[0075] Comparative Example 5

[0076] Commercially available chicken processed into minced meat, and processed according to the heating treatment method of Example 7.

[0077] Experimental Example 1 Flavor detection of examples and comparative examples

[0078] Cut the artificial meat prepared in Examples 1-6 and Comparative Examples 1-5 above into pieces with a size of 3 cm × 2 cm × 1 cm, evenly coat the surface with salad oil, and then put it into a preheated oven at 180 °C and bake for 10 minutes (simulating cooking).

[0079] Use an electronic nose and GC-MS / MS (Thermo Fisher model Trace 1600TSQ9610 in the United States) to analyze the flavor characteristics and flavor substances of the above materials. The analysis conditions are as follows:

[0080] 1. Electronic nose (Airsense model PEN3 in Germany)

[0081] Method: Weigh 3 g of the above processed materials respectively and place them in a special cup for the electronic nose. After sealing, place them at room temperature for 30 minutes to balance, and then insert the injection needle to start the measurement;

[0082] Sampling conditions: Sampling time is 1 s / group; Sensor self-cleaning time is 120 s; Sensor zeroing time is 10 s; Sample preparation time is 5 s; Injection flow rate is 400 ml / min; Analysis sampling time is 80 s. The electronic nose has ten sensors, and the numbers and functions are as follows: 1. W1C detects aromatic compounds and benzene; 2. W5S is sensitive to nitrogen oxides; 3. W3C detects ammonia in aromatic components; 4. W6s detects hydrides; 5. W5C detects short-chain alkanes and aromatics; 6. W1S detects methyl groups; 7. W1W is sensitive to sulfides; 8. W2S is sensitive to aldehydes and ketones; 9. W2W detects aromatic components and organic sulfides; 10. W3S is sensitive to long-chain alkanes.

[0083] As Figure 1 shown, the principal component analysis algorithm (PCA) shows that after adding palmitoleic acid (Example 2) and substances containing palmitoleic acid components (Examples 1, 3, 4, 5) to the material, there are significant differences compared with the blank control (Comparative Example 1). Figure 1 It can be seen from

[0084] that for Examples 1, 2, 3, and 4, the main components of their flavor substances are closer to beef (Comparative Example 3), pork (Comparative Example 4), and chicken (Comparative Example 5), and adding hemoglobin (Comparative Example 2) can produce the same effect.

[0085] As Figure 2 shown in the electronic radar chart, compared with beef (Comparative Example 3), the substances detected by the W3S, W5S, W1S, W1W, and W2S detectors in the blank control (Comparative Example 1) are all significantly insufficient, and these substances become richer than before after adding hemoglobin. Similarly, adding palmitoleic acid (Example 3) has the same effect.

[0086] 2. GC-MS / MS analysis (GC-MS / MS model Thermofish, TSQ 8000Evo)

[0087] Method: Take 3 g of the above-treated material as samples and put them into 20 ml headspace vials, seal them quickly, place the headspace vials in a 60°C constant temperature water bath, insert the solid-phase extraction injection needle into the headspace vial, extract for 1 h in the 60°C water bath and then take out, and immediately insert it into the injection port of the gas chromatography-mass spectrometry instrument for 3 min of desorption.

[0088] Gas chromatography conditions: The chromatographic column was a TG-5MS (30 m x 0.25 mm x 0.25 μm) elastic quartz capillary column. The carrier gas was high-purity helium (purity 99.999%). The carrier gas flow rate was 1.2 ml / min. Splitless injection was used. The injection port temperature was 250 °C. The temperature program was as follows: The initial temperature was maintained at 35 °C for 2 min, then increased to 180 °C at a rate of 4 °C / min and held for 2 min, and then increased to 280 °C at a rate of 15 °C / min and held for 1 min.

[0089] Mass spectrometry conditions: The ion source was an EI source. Transfer line temperature: 280 °C; Ion source temperature: 300 °C; Electron energy: 70 eV; Scanning range (m / z): 35 - 400 amu. The full scan acquisition mode was used. The mass spectrometry analysis results showed (as Figure 3 shown) that compared with the blank control (Comparative Example 1), the unique flavor volatile substances in the materials with the addition of palmitoleic acid (Example 2) and palmitoleic acid-containing components (Examples 3 - 5), and the addition of hemoglobin (Comparative Example 2) were octanal (with fruity and citrus scents), peach aldehyde / myristaldehyde (2(3H)-Furanone, 5-heptyldihydro-: fruity and buttery scents), and 2-methoxy-4-vinylphenol (2-methooxy-4-vinvylphenol: apple, peanut, and curry scents).

[0090] The mass spectrometry analysis results also showed that, compared with the blank control (Comparative Example 1), the addition of palmitoleic acid (Example 2) and components containing palmitoleic acid (Examples 3-5), and the addition of hemoglobin (Comparative Example 2) could promote the release of the following (all or most) volatile flavor substances: hexanal (which produces a grassy aroma), 1-octen-3-ol (which produces a mushroom aroma), 2-pentyl furan (which has a bean, fruity and green aroma), nonanal (which produces citrus and oily odors), maltol (which has a sweet, caramel flavor), octanoic acid (which has a fruity sour flavor), decanal (which has a sweet citrus peel and a citrusy, fatty floral aroma), (E,E)-2,4-nonadienal (which has a fresh cucumber-like aroma), nonanoic acid (which has a cheesy aroma), (E,E)-2,4-decadienal (which has a roasted chicken and oily flavor), (E,Z)-2,4-decadienal (which has a fried flavor), (E)-4-decenal (which has an orange and chicken aroma), (E)-tetradec-2-enal (which has a lemon and waxy odor), (E)-2-undecenal (which has a fruity aroma and a waxy odor).

[0091] Figure 4 The ratio of the volatile amounts of the above flavor substances in different treatments to that in Comparative Example 1 was shown. Compared with the treatments with the addition of palmitoleic acid (Example 2) and components containing palmitoleic acid (Examples 3-5), and the addition of hemoglobin (Comparative Example 2), the addition of olive oil, which is also a commonly used flavor oil (Example 6), only enhanced the volatility of a few of the above substances.

[0092] Experimental Example 2 Gastrointestinal Simulation Experiment

[0093] In vitro gastrointestinal simulation experiments mimic the human intestinal environment, especially the colon, through in vitro fermentation to study the fermentation process of intestinal microorganisms on different substrates (such as dietary fiber and prebiotics) and evaluate their health benefits. The main steps of this experiment include preparing the fermentation vessel, adding fecal inoculum, maintaining anaerobic conditions, adding the substrate to be tested, sampling regularly, and monitoring changes in the microbial composition. Through metagenomics analysis based on next-generation sequencing, the abundances and functions of different intestinal microorganisms are predicted. Metagenomics technology can provide detailed information on the microbial community structure and reveal the impact of substrates on the intestinal microbial community. This experimental method can not only demonstrate the promoting effect of substrates on beneficial bacteria (such as Bifidobacterium and Lactobacillus), but also evaluate their inhibitory effect on pathogenic bacteria (such as Escherichia coli and Clostridium perfringens). In addition, by analyzing microbial metabolites, the metabolic pathways of substrates in the intestine and their potential health benefits can be further understood. Gastrointestinal simulation experiments provide a controllable platform for studying the relationship between dietary components and intestinal health, helping to develop functional foods and dietary supplements to improve human health. In summary, gastrointestinal simulation experiments combined with metagenomics analysis can deeply analyze the interaction between intestinal microorganisms and dietary components, providing important data support for predicting the possible effects of foods or food ingredients on human digestion.

[0094] 1. In vitro fermentation

[0095] To explore the digestion, decomposition, and microbial interactions of the examples in a controlled simulation environment, the present invention refers to the in vitro fermentation method in existing literature (Li, H. et al. In vitro fermentation of human milk oligosaccharides by individual Bifidobacterium longum-dominant infant fecal inocula. Carbohydrate Polymers 287, 119322 (2022)). Nine healthy volunteers aged 24 - 32 years with normal body mass index (BMI) were selected for the experiment, and their fresh fecal samples were collected. The volunteer screening criteria were no history of digestive tract diseases and no history of antibiotic use in the past three months. All participants signed informed consent forms, and the experimental protocol was approved by the Medical Ethics Committee of Nanjing Integrated Traditional Chinese and Western Medicine Hospital (batch number:

[0096] S2021 - 10 - 008).

[0097] To prepare samples for in vitro fermentation, the collected fecal material was mixed and diluted with phosphate buffer solution (PBS) at pH 7.2 to make a 10% fecal suspension (w / v). Then the suspension was homogenized and filtered through four layers of sterile gauze to ensure uniformity and remove larger particles. An intestinal simulation reactor (BGR-03, Jiangnan University, Wuxi, Jiangsu, China) was used to simulate the human digestive environment. To simulate the anaerobic conditions of the intestine, the reactor was purged with nitrogen three times before the start of fermentation. Then, 200 mL of prepared brain heart infusion (BHI) and mucin medium (including 11.1 g of BHI and 1.5 g of mucin dissolved in 1 liter of sterilized water and sterilized at 115 °C for 20 minutes) were introduced into the reactor. The fecal suspension was inoculated into the reactor and incubated for 24 hours to establish a stable microbial community and simulate the composition of the human gut microbiota. After the initial 24-hour stabilization period, 5 mL of phosphate buffer solution containing finely ground vacuum-dried samples (Example 1 (containing sea buckthorn juice), Example 3 (containing palmitoleic acid), Comparative Example 1 (blank), Comparative Example 3 (beef), and a plant-based meat purchased on the market, Beyond ) was injected into the reactor and maintained at 37 °C to simulate physiological conditions. Samples were collected at 0, 6, 12, 18, and 24 hours to monitor microbial activity and metabolic changes. Each collected sample was centrifuged at 10,000 rpm for 5 minutes, and the resulting precipitate was frozen and stored at -80 °C for further analysis.

[0098] 2. Intestinal microbiota analysis

[0099] The microbiota analysis of fecal samples included multiple steps: First, genomic DNA was extracted from 24-hour fermentation samples and the control group (without adding any meat or plant-based meat samples, only brain heart infusion (BHI) and mucin medium) using the TIANamp Stool DNA Extraction Kit (Tiangen Biochemical, Beijing), and subsequent sequencing analysis was completed by Novogene Bioinformatics Technology Co., Ltd., Beijing. The V3-V4 region of the 16S rRNA gene was PCR amplified using primers F: 5’-CCTAYGGGRBGCASCAG-3’ and R: 5’-GGACTACNNGGGTATCTAAT-3’, and the amplified products were sequenced on the NovaSeq6000 platform (Illumina, USA). Bioinformatics analysis started from processing the valid sequencing data, clustering the sequences into operational taxonomic units (OTUs) with 97% similarity, and generating a normalized OTU abundance table for subsequent analysis.

[0100] The results showed that Example 1 had a similar diversity of intestinal microbiota to Comparative Example 3 (such as Figure 5as shown). Moreover, compared with other test samples, Example 1 and Example 3 significantly inhibited the proliferation of Escherichia-Shigella. The Escherichia-Shigella flora is generally considered to contain a variety of harmful and pathogenic bacteria among gut microbiota (such as Figure 6 as shown, marked with asterisks in the figure).

[0101] Functional prediction of gut microbiota by TAX4FUN showed that both Example 1 and Example 3 could positively regulate the functions of gut microbiota, specifically involving carbohydrate metabolism, glycan biosynthesis and metabolism, endocrine and metabolic diseases, as well as functions related to the endocrine, nervous, and immune systems ( Figure 7 marked with asterisks in

[0102] This invention fully demonstrates that palmitoleic acid and natural substances rich in it can be used as good seasonings for simulating meat flavors, applied to the processing of plant-based meat, artificial meat, and related products, and provide additional health benefits.

Claims

1. An artificial meat, characterized in that: The artificial meat contains more than 0.01% by mass of palmitoleic acid, and preferably, the mass percentage of the palmitoleic acid is 0.01-0.07%.

2. The artificial meat according to claim 1, characterized in that: The palmitoleic acid in the artificial meat is derived from natural or genetically engineered biological products rich in palmitoleic acid. Preferably, the natural products rich in palmitoleic acid include one or more of sea buckthorn juice, sea buckthorn oil, macadamia oil or avocado oil; preferably, the genetically engineered biological products rich in palmitoleic acid include palmitoleic acid-containing products produced by engineered bacteria or plants optimized and constructed through gene mutagenesis, gene editing or transgenic technology.

3. The artificial meat according to claim 1, characterized in that: The artificial meat also includes a main ingredient and auxiliary ingredients. Preferably, the main ingredient includes a base composed of a plant protein matrix, and the auxiliary ingredients include natural products of other plants or yeast and natural flavors.

4. The artificial meat according to claim 3, characterized in that: The plant protein matrix includes one or more of soy protein, wheat protein or potato protein. Preferably, the auxiliary materials include one or more of resistant dextrin, gluten powder, beet powder, mushroom essence, sunflower oil, coconut oil or yeast extract.

5. The artificial meat according to claim 1, characterized in that: The artificial meat is made of the following components in parts by weight: 0.1-0.7 parts of palmitic acid, 200-210 parts of soy protein, 20-25 parts of potato protein, 8-10 parts of resistant dextrin, 100-130 parts of gluten, 16-18 parts of beet powder, 36-40 parts of yeast extract, 6-7 parts of mushroom essence, 24-25 parts of rapeseed oil, 18-20 parts of coconut oil and 0.2-0.25 parts of salt.

6. The method for preparing artificial meat according to any one of claims 1 to 5, characterized in that: The following steps are involved: 1) mixing part of the raw materials and / or part of the raw material pretreatment products to obtain a premix; 2) adding the premix to other raw materials and mixing them evenly to obtain a vegetable meat mixture, wherein the other raw materials include natural or genetically engineered biological products rich in palmitoleic acid; Preferably, the preparation method further comprises placing the plant meat mixture described in step 2) into a mold and compacting it with appropriate pressure.

7. The method for preparing artificial meat according to claim 6, characterized in that: The preparation method specifically comprises the following steps: 1) Accurately weigh the following ingredients: soy protein, potato protein, resistant dextrin or resistant starch, gluten, beet powder, yeast extract, mushroom extract, sunflower oil, palmitoleic acid or natural products rich in palmitoleic acid, organic coconut oil and salt; 2) Soy protein pretreatment: Soak the soy protein in hot water, keep the temperature at 95-100℃, and ensure that it fully absorbs water. After soaking, drain it with a sieve; 3) Mixing and homogenizing ingredients: Mix the above-mentioned pretreated soy protein, gluten, rapeseed oil and coconut oil to ensure that the gluten and soy protein are mixed, and the oil is fully mixed to obtain a premix; place the premix together with other raw materials in a mixer; mix at a controlled speed and time until a homogeneous and consistent plant meat mixture is obtained; preferably, the speed is 120-150rpm and the time is 10-15min.

8. A method for seasoning artificial meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid, characterized in that: The artificial meat seasoning method comprises the following steps: adding palmitoleic acid or a natural or genetically engineered biological product containing palmitoleic acid into a main vegetable protein matrix material and other auxiliary materials for heating treatment.

9. The method for seasoning artificial meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid according to claim 8, characterized in that: The content of the palmitoleic acid is 0.01-0.07%.

10. The method for seasoning artificial meat based on palmitoleic acid or natural or genetically engineered biological products containing palmitoleic acid according to claim 8, characterized in that: The heating temperature is 150-180° C., and the heating time is 15-20 minutes.

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