A method for improving color and flavor of plant-based meat products using bovine myoglobin

Abstract

The application discloses a method for improving color and flavor of plant-based meat products by using bovine myoglobin, and belongs to the field of food biotechnology and plant-based meat product processing. In view of the problems of pale color and lack of flavor of plant-based meat products, soybean protein isolate is prepared into a water suspension, and particulate matter is removed by centrifugation. Bovine myoglobin or cell lysate containing bovine myoglobin is added to the centrifuged water suspension, mixed uniformly, and then subjected to heat treatment at 95 DEG C after standing at room temperature, and then cooled to obtain a plant-based meat product. The plant-based meat product prepared by the method has a color changed from light yellow to meat-like red, and the content of grassy substances such as hexanal is reduced, the content of roasted, floral and mushroom characteristic flavor substances such as 2-pentyl furan, nonanal, 1-octene-3-ol and benzaldehyde is increased, and the flavor is significantly improved. The method solves the problems of poor color and flavor of plant-based meat products, and the use of cell lysate containing bovine myoglobin can reduce the application cost.

Description

Technical Field

[0001] This invention relates to a method for improving the color and flavor of plant-based meat products using bovine myoglobin, belonging to the fields of food engineering and enzyme engineering technology. Background Technology

[0002] With consumers' growing awareness of health and environmental protection, the plant-based meat market is experiencing rapid growth. These products typically use plant proteins such as soy protein isolate and pea protein as main raw materials, mimicking the fibrous texture of animal muscle through processing methods such as extrusion and heat-induced gelation. However, due to the inherent characteristics of plant proteins and limitations in processing technology, existing plant-based meat products still show significant differences in color and flavor compared to real meat.

[0003] Myoglobin is a heme-binding protein composed of globin and heme prosthetic groups. In animal muscle tissue, it is not only the main source of red color but also a key factor that gives meat its unique "meaty" flavor. Plant-based meat products, lacking animal muscle tissue, naturally lack this component, which is the fundamental reason for their pale color and bland flavor.

[0004] Bovine myoglobin's advantage lies in its ability to provide a "full sensory simulation" that goes beyond simple coloring. In terms of color, it gives plant-based meat a red hue similar to real meat and undergoes a color change consistent with real meat during cooking. In terms of flavor, it can participate in reactions such as lipid oxidation and thermal degradation to generate volatile flavor compounds such as aldehydes and ketones, creating a unique "meaty" taste. This dual function is difficult for other additives to replace, making it an ideal choice for improving the quality of plant-based meat products. Therefore, providing a method for improving the color and flavor of plant-based meat products using bovine myoglobin obtained from Bacillus subtilis fermentation is of great significance for improving the quality and promoting the market of plant-based products. Summary of the Invention

[0005] This invention adds bovine myoglobin to meat products, giving them the color and flavor characteristics of real meat, thus optimizing the sensory quality of plant-based meat products in terms of both sight and smell.

[0006] This invention provides a method for preparing plant-based meat products with enhanced sensory quality, comprising the following steps: (1) Mix the plant protein raw materials with water to obtain a plant protein suspension; (2) Add bovine myoglobin or cell lysate containing bovine myoglobin to the plant protein suspension prepared in step (1) and mix well; (3) Allow the mixture from step (3) to stand. (4) Heat-treat the material after step (4) and then cool it to obtain plant-based meat products.

[0007] In one embodiment, the plant protein raw material is soy protein isolate.

[0008] In one embodiment, the bovine myoglobin is added in the form of a purified protein.

[0009] In one embodiment, the bovine myoglobin is added in the form of a cell lysate containing the protein.

[0010] In one embodiment, the bovine myoglobin is obtained by microbial fermentation.

[0011] In one embodiment, the amount of bovine myoglobin added is 1 to 5% of the total mass of the plant protein raw materials.

[0012] In one embodiment, the heat treatment temperature is 90~95℃ and the treatment time is 15~30 min.

[0013] The present invention also provides plant-based meat products prepared using the method described herein.

[0014] This invention also provides the application of bovine myoglobin in improving the color and / or flavor of plant-based meat products.

[0015] In one implementation, the improvement in color includes increasing the redness value a and / or decreasing the lightness value L.

[0016] In one embodiment, the flavor improvement includes reducing the content of hexanal and / or increasing the content of at least one of 2-pentylfuran, nonanal, 1-octen-3-ol, and benzaldehyde.

[0017] The beneficial effects of this invention are: This invention provides a method for improving the color and flavor of plant-based meat products. By adding bovine myoglobin in the form of purified protein or cell lysate to soy protein isolate, and then heat-treating, the color of the plant-based meat product changes from pale yellow to a flesh-colored hue, exhibiting a good coloring effect. Simultaneously, this invention improves the flavor profile of plant-based meat products, reducing the content of grassy flavor compounds such as hexanal, and increasing the content of characteristic flavor compounds such as roasted, floral, and mushroom aromas, such as 2-pentylfuran, nonanal, 1-octen-3-ol, and benzaldehyde, resulting in a richer and more pleasant flavor that more closely resembles the sensory characteristics of real meat. This effectively solves the problem of pale color and monotonous flavor in plant-based meat products.

[0018] The method of this invention is simple, and the direct use of cell lysate containing bovine myoglobin can significantly reduce application costs, thus showing good application prospects. Attached Figure Description

[0019] Figure 1Color characterization of different groups of plant-based meat products; where control is plant-based meat product prepared without the addition of B-Mb; lB-Mb is B-Mb in cell lysate; pB-Mb is purified B-Mb; and sH-Mb is standard H-Mb. Detailed Implementation

[0020] Soy protein isolate Suyun 626 (SPI) was purchased from Lianyungang Fengyuan Seed Industry Technology Co., Ltd. After neutralization to pH 7.0, the protein content of SPI was determined by the Kjeldahl method, and the result was 92% on a dry weight basis. SPI was prepared into a 10% (w / v) deionized water suspension and centrifuged at 10,000 g for 20 min to remove particulate matter.

[0021] The method for preparing purified B-Mb protein was based on the paper "Metabolic engineering of Bacillus subtilis for the production of active hemoglobins and myoglobins by improving heme supply," which used the engineered Bacillus subtilis Heme-K2 strain as the host, carrying the pMINI-gntR-B-Mb plasmid (sodium gluconate-induced, high copy number). The strain was cultured in BSM medium at 37°C and 220 rpm. Six hours after inoculation, sodium gluconate was added to induce expression, and the cells were harvested 24 hours later. The cells were resuspended in PBS buffer (pH 7.4) and then homogenized using an autoclave. The homogenized mixture was centrifuged at 4°C and 10,000 rpm for 15 min, and the supernatant was collected. Since B-Mb has a 6×His tag at its C-terminus, affinity chromatography was performed using the BeaverBeads™ His-tagged protein purification system (nickel column). Finally, the purified B-Mb (pB-Mb) was obtained by desalting and concentrating the protein using an Amicon Ultra 10 K ultrafiltration centrifuge tube. The preparation method of cell lysate containing B-Mb protein is based on "Metabolic engineering of Bacillus subtilis for the production of active hemoglobins and myoglobins by improving heme supply". After high-pressure homogenization and centrifugation, the supernatant of Bacillus subtilis Heme-K2 engineered strain expressing B-Mb can be collected directly for use.

[0022] Example 1 (1) Raw material pretreatment Crush and sieve the soy protein isolate (SPI) for later use. Store bovine myoglobin (B-Mb) or cell lysates containing B-Mb at 4°C for later use.

[0023] (2) Preparation of plant-based meat product samples Soy protein isolate was prepared into a 10% (w / v) deionized water suspension and centrifuged at 10,000 g for 20 min to remove particulate matter.

[0024] Bovine myoglobin, either in purified protein form or in the form of cell lysate containing the protein, was added at 1% of the total mass of the plant protein raw material. The mixture was stirred for 20 minutes to mix evenly, allowed to stand at room temperature for 20 minutes, and then heat-treated at 95°C for 20 minutes. Subsequently, it was cooled to room temperature in an ice-water bath to obtain a sample of plant-based meat products.

[0025] (3) Evaluation of coloring effect After the heat-treated samples were cooled to room temperature, the surface color parameters (L, a, b) were measured using a colorimeter.

[0026] As shown in Table 1, the redness value (a) of the sample with added purified B-Mb increased from 0.99 to 3.24, the lightness value (L*) decreased from 40.55 to 36.37, and the color changed from pale yellow to flesh-colored. The sample with added B-Mb cell lysate also showed good staining effect and did not introduce any unpleasant odor, which verified the feasibility of directly using B-Mb-containing cell lysate for addition.

[0027] Table 1. Color evaluation of SPI samples with the addition of Mb

[0028] (4) Flavor assessment Measurements were performed using a single quadrupole gas chromatography-mass spectrometry (GC-MS) system equipped with an SH-WAX column (30 m × 0.25 mm × 0.25 μm). The injection port was run at 250°C in splitless mode for 7 min, using helium as the carrier gas at a flow rate of 1.8 mL / min. The column oven program started at 40°C, held for 0.5 min, then increased to 100°C at a rate of 5°C / min, followed by acceleration to 230°C at a rate of 10°C / min, and finally held at 230°C for 5 min. Flavor compounds were separated by the capillary column and split at a 1:1 (v / v) ratio to the mass spectrometer (MS) and olfactometer. The MS ion source temperature was maintained at 200°C, while the interface temperature was set to 250°C. Ions with a mass-to-charge ratio (m / z) ranging from 33 to 400 were detected using a full-scan mode, operated in electron impact (EI) mode with an ionization energy of 70 eV. The olfactory detector's transmission line temperature was maintained at 200°C, and humidified air was supplied at a flow rate of 40 mL / min to mitigate the drying effect. Trained evaluators assessed the emitted aromas using ODE-2030 software, recording their flavor descriptions and intensities. A panel of three skilled GC / O evaluators (two men and one woman) systematically evaluated flavor characteristics, intensity, and aroma pleasantness every other day for two weeks. Odor characteristics were analyzed using the Odor Specificity Intensity Assessment (OSME) method, with intensity scores recorded on a scale of 0 to 5. The intensity of odor extracts was recorded, and each panel member re-evaluated each sample to ensure consistency and accuracy.

[0029] As shown in Table 2, the content of grassy flavor substances such as hexanal decreased in the samples, while the content of characteristic flavor substances such as roasted aroma, floral aroma, and mushroom aroma, such as 2-pentanone, 2-pentylfuran, 3-hydroxybutanone, nonanal, 1-octen-3-ol, and benzaldehyde, increased, and the flavor profile was significantly improved.

[0030] Table 2. Identification of odor-active compounds in SPI samples after B-Mb addition

[0031] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for preparing plant-based meat products with enhanced sensory quality, characterized in that, Includes the following steps: (1) Mix the plant protein raw materials with water to obtain a plant protein suspension; (2) Add bovine myoglobin or cell lysate containing bovine myoglobin to the plant protein suspension prepared in step (1) and mix well; (3) Allow the mixture from step (2) to stand; (4) Heat-treat the material after step (3) and then cool it to obtain plant-based meat products.

2. The method according to claim 1, characterized in that, The plant protein raw material is soy protein isolate.

3. The method according to claim 1, characterized in that, The bovine myoglobin was added in the form of purified protein.

4. The method according to claim 1, characterized in that, The bovine myoglobin was added in the form of a cell lysate containing the protein.

5. The method according to any one of claims 1 to 4, characterized in that, The amount of bovine myoglobin added is 1-5% of the total mass of the plant protein raw materials.

6. The method according to any one of claims 1 to 5, characterized in that, The heat treatment temperature is 90~95℃, and the treatment time is 15~30 min.

7. Plant-based meat products prepared by the method described in any one of claims 1 to 6.

8. Application of bovine myoglobin in improving the color and / or flavor of plant-based meat products.

9. The application according to claim 8, characterized in that, The improvement in color includes increasing the redness value a and / or decreasing the lightness value L.

10. The application according to claim 8, characterized in that, The improved flavor includes reducing the content of hexanal and / or increasing the content of at least one of 2-pentylfuran, nonanal, 1-octen-3-ol, and benzaldehyde.

Images