Beef myosin flavor peptide CM1 and application thereof

Abstract

The invention discloses a beef myosin flavor peptide CM1, the amino acid sequence of the beef myosin flavor peptide CM1 is EDMAE, and the molecular weight of the flavor peptide is 594 Da. The flavor peptide belongs to umami peptide, the umami threshold value of the flavor peptide is smaller than the umami threshold value of monosodium glutamate, the flavor peptide has good umami and umami enhancing characteristics, the umami characteristics of monosodium glutamate, soy sauce and soup can be remarkably improved, the flavor peptide can be used as a novel seasoning for replacing monosodium glutamate, the umami and mellowness of the whole taste of food are improved, and the flavor peptide has good market application prospects.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a beef myosin flavor peptide CM1 and an application thereof. Background Art

[0002] With the in-depth study of bioactive peptides, many functional peptides such as lowering blood pressure, lowering blood lipids, and presenting flavor have been reported. Among them, flavor peptides have attracted widespread attention from many researchers because they can produce distinct characteristic tastes in both natural foods and processed foods. Flavor peptides are divided into five categories according to the basic taste they present: sweet peptides, sour peptides, bitter peptides, salty peptides, and umami peptides. Flavor peptides, as a type of polypeptide, can improve the freshness of food, enhance food taste perception, and increase consumer appetite. Due to their high taste characteristics and physiological activity, they are gradually used in high-end condiments and are a food seasoning product with market potential.

[0003] Umami peptides, as a flavor peptide that enhances the umami and mellow taste of food, have good processing characteristics and freshness effects. Moreover, they can not only directly enhance the taste and flavor of food, but also participate in the Maillard reaction as a precursor of volatile flavor substances, further enhancing the characteristic flavor of food, which is in line with the "natural and healthy" dietary trend and consumer psychology. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art, provide a beef myosin flavor peptide CM1 and its application, and solve the problems in the above-mentioned background technology.

[0005] One of the technical solutions adopted by the present invention to solve the technical problem is: providing beef myosin flavor peptide CM1, whose amino acid sequence is EDMAE, as shown in SEQ ID NO:1.

[0006] Studies have shown that the molecular weight of umami peptides is usually below 3000Da, and the amino acid composition of umami peptides is often expressed as glutamic acid, aspartic acid, asparagine, glutamine combined with each other or combined with threonine, serine, alanine, glycine, methionine and cysteine ​​to form polyacid sodium salts. At the same time, the sweet amino acids threonine, glycine, serine and alanine account for a high proportion. There is a theoretical basis for finding flavor peptides with this as the target. We first screened and calculated the beef myosin sequence and found the umami sequence EDMAE, named CM1, with a molecular weight of 594Da.

[0007] Practical applications have confirmed that flavor peptide CM1 can be used in braised soup and meat jerky to replace MSG as a new type of umami seasoning.

[0008] The second technical solution adopted by the present invention to solve the technical problem is: providing the above-mentioned beef myosin flavor peptide CM1 related biological material, which is any one of the following A1) to A8):

[0009] A1) a nucleic acid molecule encoding the recombinant protein according to claim 1;

[0010] A2) an expression cassette containing the nucleic acid molecule described in A1);

[0011] A3) a recombinant vector containing the nucleic acid molecule described in A1),

[0012] A4) a recombinant vector containing the expression cassette described in A2);

[0013] A5) a recombinant microorganism containing the nucleic acid molecule described in A1);

[0014] A6) a recombinant microorganism containing the expression cassette described in A2);

[0015] A7) a recombinant microorganism containing the recombinant vector described in A3);

[0016] A8) A recombinant microorganism containing the recombinant vector described in A4).

[0017] The third technical solution adopted by the present invention to solve its technical problem is: to provide the use of the above-mentioned beef myosin flavor peptide CM1 in the preparation of food flavoring or health care products.

[0018] The fourth technical solution adopted by the present invention to solve its technical problem is: to provide the use of the above-mentioned beef myosin flavor peptide CM1 related biological materials in the preparation of food flavorings or health products.

[0019] The fifth technical solution adopted by the present invention to solve its technical problem is: to provide a food seasoning, whose effective ingredient includes beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO: 1.

[0020] Preferably, the active ingredient of the above-mentioned food flavoring agent is beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:1.

[0021] The sixth technical solution adopted by the present invention to solve its technical problem is: to provide a health product, whose effective ingredient includes beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO: 1.

[0022] Preferably, the active ingredient of the beef myosin flavor peptide CM1 is beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:1.

[0023] The flavor peptides of the present invention can be synthesized by methods known to those skilled in the art, such as solid phase synthesis, and purified by methods known to those skilled in the art, such as high performance liquid chromatography.

[0024] The implementation of the present invention has the following beneficial effects:

[0025] The present invention takes beef myosin as the research object, and through screening and calculation, finds a polypeptide CM1 with a completely new amino acid sequence. The flavor peptide CM1 presents a strong umami taste, can be used as a substitute for monosodium glutamate, and can replace monosodium glutamate in braised stewed soup and meat jerky, presenting a better flavor in the braised stewed soup and meat jerky. The flavor peptide CM1 of the present invention has a good application prospect in the production of food seasonings or health products. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the structure of flavor peptide CM1.

[0027] Figure 2 This is a picture of the flavor peptide CM1 compound freshness enhancement effect.

[0028] Figure 3 This is the flavor radar chart of flavor peptide CM1.

[0029] Figure 4 This is a homology simulation diagram of flavor receptor T1R1 / T1R3.

[0030] Figure 5 Schematic diagram of the 3D structure of flavor peptide CM1.

[0031] Figure 6 Schematic diagram of the molecular docking results of flavor peptide CM1 and umami taste receptor T1R1 / T1R3. DETAILED DESCRIPTION

[0032] In order to better understand the present invention, the present invention is further described in detail below in conjunction with the embodiments and drawings. However, those skilled in the art understand that the following embodiments are not limitations on the scope of protection of the present invention, and any changes and modifications made on the basis of the present invention are within the scope of protection of the present invention.

[0033] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0034] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0035] Example 1 Screening process of beef myosin flavor peptide CM1

[0036] Beef myosin was used as the research object, and the UniProt (https: / / www.uniprot.org / ) protein database was used for retrieval. Some amino acid sequences rich in hydrophilic amino acids (such as aspartic acid and glutamic acid) in the myosin sequence were screened, and the umami taste was predicted by virtual screening methods, including UMPred-FRL (https: / / pmlabstack.pythonanywhere.com / UMPred-FRL), Umami-YYDS (http: / / www.tastepeptides-meta.com / Umami_YYDS), and TastePeptides DM (http: / / tastepeptides-meta.com / TPDM). The sequences predicted as umami by the three online prediction servers at the same time were considered to be potential umami peptides.

[0037] Three online prediction servers predicted that EDMAE has umami taste, so this sequence was selected as a potential umami peptide for subsequent experiments. Figure 1 shown.

[0038] Example 2 Sensory Verification of Flavor Peptide CM1

[0039] The synthetic flavor peptide CM1 solution was used as the sample solution for sensory evaluation. 0.35% salt solution, 1% sucrose solution, 0.08% citric acid solution, 0.35% monosodium glutamate solution, and 0.08% quinine solution were used as the basic solutions for the five tastes of salty, sweet, sour, fresh, and bitter, respectively. The basic solution was scored as 5 points, and the target flavor solution was scored on a 10-point scale to evaluate the flavor intensity of the flavor peptide CM1. The sensory evaluation was conducted by a well-trained professional sensory evaluation team (half male and half female). Each evaluation sample was randomly numbered and shuffled to the evaluator for taste evaluation. Each evaluator evaluated the same sample 3 times, and the average of the 3 measurements was taken as the final evaluation score. Each sample was evaluated at room temperature. Each sensory officer was also required to evaluate the taste characteristics of each sample presented. The sensory scoring data was analyzed using EXCEL, and the final sensory evaluation results are shown in Table 1.

[0040] Table 1 Taste characteristics of peptide CM1

[0041]

[0042] Example 3 Electronic tongue evaluation of flavor peptide CM1

[0043] First, prepare the synthesized solution into 1 mg / mL. Next, accurately transfer 25 mL of the test solution into the sample cup dedicated to the electronic tongue. Before measurement, perform self-test, activation, calibration and diagnosis on the electronic tongue to ensure the reliability and stability of the collected data. Use 30 mmol / L KCl solution and 0.3 mmol / L tartaric acid solution to prepare the Ref Sol reference solution. Place the sensor in the reference solution and return to zero for 30 seconds, then start the umami measurement. The test time is 30 seconds. After the test is completed, wash with the reference solution for 3 seconds, and then perform the aftertaste measurement again, and the test time is 30 seconds. Each sample is repeated 4 times, and the last 3 times are taken as the test result. The test results are shown in Tables 2 and Figure 3 As shown. Figure 3 It can be seen that the flavor peptide CM1 has more obvious fresh, salty and sweet tastes, and weaker bitter and sour tastes. The public prefers fresh, salty and sweet tastes, and has a lower acceptance of bitter and sour tastes, indicating that the umami peptide can significantly improve the taste of food and has broad application prospects in improving the freshness of food.

[0044] Table 2 Electronic tongue identification results 6 taste response values

[0045] serial number Umami sour taste bitterness Astringency Salty Sweetness Aftertaste Richness 1 10.07 0.48 -0.09 1.10 5.01 1.01 2.49 2.78 1 10.07 0.48 -0.09 1.10 5.01 1.01 2.49 2.78 1 10.07 0.48 -0.09 1.10 5.01 1.01 2.49 2.78

[0046] Example 4: Composite flavor enhancement effect of flavor peptide CM1

[0047] In order to explore the composite flavor enhancement effect of umami peptide CM1, the synthetic umami peptide CM1 was prepared into a 0.5 mg / mL solution, and the composite solution of umami peptide CM1 was prepared by the mode of 2 mg / mL sodium glutamate + 2 mg / mL sodium chloride + synthetic peptide to evaluate the synergistic flavor enhancement effect of umami peptide. The umami and salty taste of the umami peptide composite solution were compared with the single CM1+NaCl solution through sensory evaluation to analyze the synergistic flavor enhancement effect of umami peptide CM1. The scoring method refers to the sensory evaluation method in Example 1. The composite umami solution CM1+NaCl was identified as 5 points, and the umami and saltiness of the composite umami peptide sample solution were scored on a 10-point system, with 0 points representing no taste and 10 points representing significant taste. The final composite synergistic flavor enhancement results are shown as follows. Figure 2 shown.

[0048] Example 5 Molecular docking of flavor peptide CM1 with umami taste receptor

[0049] Umami peptides can show certain umami characteristics by interacting with umami taste receptors, so umami receptors are considered to be the key to perceiving umami. The umami receptors discovered so far mainly include heterodimers T1R1 / T1R3, which have a higher affinity for umami substances and are the main umami receptors. In order to further study the taste characteristics and taste mechanism of umami peptides, homology modeling and molecular docking can be used to explain the interaction between umami peptides and taste receptors.

[0050] 1. Homology Modeling of Umami Taste Receptors

[0051] The 3D structure of the umami receptor T1R1 / T1R3 can be constructed by homology modeling. The amino acid sequence of T1R1 / T1R3 can be obtained from the UniProt (https: / / www.uniprot.org / ) database, and then a model with metabolic glutamate as the receptor template can be constructed using SWISS-MODEL (https: / / swissmodel.expasy.org / ). Figure 3 shown.

[0052] 2. Molecular docking of umami peptides with umami receptors T1R1 / T1R3

[0053] The 3D structure of the umami peptide CM1 was constructed and optimized by the I-TASSER (https: / / zhanggroup.org / ) online server, removing all water molecules and redundant components, and adding hydrogen atoms and force fields. The identified umami peptide was docked into the pocket of T1R1 / T1R3 through molecular visualization. The stability of the docked complex was estimated. The better the stability, the better the binding effect of the small molecule peptide with the umami receptor. The docking results of the umami peptide molecule and the umami receptor molecule are shown in the figure. Figure 4 , 5 shown.

[0054] Example 6 Practical application of flavor peptide CM1 in braised soup and meat jerky

[0055] Ingredients and proportions of the braised soup recipe: 80 parts of pork bones, 2 parts of rock sugar, 1 part of oyster sauce, and 1 part of flavor peptide.

[0056] Ingredients and proportions of the meat jerky formula: 60 parts of lean pork, 10 parts of pork fat, 5 parts of salt, 3 parts of five-spice powder, 5 parts of white sugar, 1.0 part of complex phosphate, and 0.03 part of flavor peptide.

[0057] Preparation method of braised soup:

[0058] Directly weigh the ingredients in the formula and cook for 4 hours to obtain the flavor peptide braised soup.

[0059] Preparation method of meat jerky:

[0060] S1. Chopping and mixing: Take lean pork, pork fat, salt, five-spice powder, white sugar, complex phosphate, and flavor peptide according to the formula, mix well, and then chop and mix to obtain minced meat;

[0061] S2. Cooking: Mechanically squeeze the minced meat into slices with a thickness of 2 to 3 mm, apply syrup on both sides of the slices, and steam;

[0062] S3. Baking: Baking the steamed meat slices to obtain flavored peptide meat jerky.

[0063] The specific baking process is as follows: the flavored peptide meat jerky is first steamed for 5 minutes, then pre-baked for 3 to 5 minutes, and then baked at 200 to 250 degrees Celsius for 5 to 10 minutes.

[0064] The same amount of monosodium glutamate was used to replace the umami peptides in the braised soup and meat jerky, and the braised soup and meat jerky were prepared in the same way. The umami peptide braised soup, meat jerky, monosodium glutamate braised soup, and meat jerky were subjected to sensory evaluation. The sensory scoring table is shown in Tables 3 and 4 below. The test method is to select five people to score from four aspects of color, aroma, taste, and mouthfeel, and calculate the final average score and total score.

[0065] Table 3 Sensory test table of braised soup

[0066]

[0067]

[0068] Table 4 Sensory evaluation results

[0069] project Umami peptide braised soup MSG Braised Soup Color 9.6 9.6 Flavor 9.6 9.5 smell 9.4 9.3 Total score 28.6 28.4

[0070] Referring to Table 3 and Table 4, it can be seen from the data analysis of Example 6 that the flavor peptide braised soup is superior to the monosodium glutamate braised soup in flavor and taste. The monosodium glutamate braised soup is only the same as the umami peptide braised soup in color. Therefore, it can be determined that the technical solution of the present invention has a technical effect that is significantly better than the traditional braised soup.

[0071] Table 5 Sensory test table of meat jerky

[0072]

[0073] Table 6 Sensory evaluation results

[0074] project Umami peptide meat jerky MSG meat jerky Color 9.3 9.4 Flavor 9.6 9.5 smell 9.4 9.3 Taste 9.4 9.3 Total score 37.7 37.5

[0075] Referring to Table 5 and Table 6, it can be seen from the data analysis of Example 6 that the flavor peptide meat jerky is superior to the monosodium glutamate meat jerky in flavor, taste and mouthfeel. The monosodium glutamate meat jerky is only slightly stronger than the umami peptide meat jerky in color. Therefore, it can be determined that the technical solution of the present invention has a technical effect that is significantly superior to the traditional monosodium glutamate meat jerky.

[0076] In summary, the beef myosin flavor peptide CM1 provided by the present invention has a strong umami taste, can be used as a substitute for monosodium glutamate, and can replace monosodium glutamate in braised soup and meat jerky, and has a good flavor in braised soup and meat jerky. The flavor peptide CM1 of the present invention has a good application prospect in the production of food seasonings or health products.

[0077] Although the specific implementation modes of the present invention are described above, those skilled in the art should understand that the specific implementation modes described are only illustrative and are not intended to limit the scope of the present invention. Equivalent modifications and changes made by those skilled in the art in accordance with the spirit of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. A beef myosin flavor peptide CM1, characterized in that: The amino acid sequence of the beef myosin flavor peptide CM1 is shown in SEQ ID NO:

1.

2. The biological material related to the beef myosin flavor peptide CM1 according to claim 1 is any one of the following A1) to A8): A1) a nucleic acid molecule encoding the recombinant protein according to claim 1; A2) an expression cassette containing the nucleic acid molecule described in A1); A3) a recombinant vector containing the nucleic acid molecule described in A1), A4) a recombinant vector containing the expression cassette described in A2); A5) a recombinant microorganism containing the nucleic acid molecule described in A1); A6) a recombinant microorganism containing the expression cassette described in A2); A7) a recombinant microorganism containing the recombinant vector described in A3); A8) A recombinant microorganism containing the recombinant vector described in A4).

3. Use of the beef myosin flavor peptide CM1 according to claim 1 in preparing food flavoring.

4. Use of the biomaterial related to the beef myosin flavor peptide CM1 according to claim 2 in the preparation of food flavorings.

5. A food flavoring agent, characterized in that: The effective component comprises beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:

1.

6. The food flavoring agent according to claim 5, characterized in that: The effective component is beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:

1.

7. A health product, characterized in that: The effective component comprises beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:

1.

8. The health product according to claim 7, characterized in that: The effective component is beef myosin flavor peptide CM1, and the amino acid sequence of the beef myosin flavor peptide CM1 is SEQ ID NO:1.

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