Salty peptide sourced from shiitake mushroom and squid soup and application of salty peptide in food seasoning

Abstract

An amino acid sequence of the salty peptide sourced from shiitake mushroom and squid soup is shown as SEQ ID NO.1, the salty peptide sourced from shiitake mushroom and squid soup is obtained through protease hydrolysis and chromatographic separation technologies, the amino acid sequence of the salty peptide sourced from shiitake mushroom and squid soup is identified through mass spectrometry, and molecular docking is carried out to evaluate the binding characteristic of the salty peptide sourced from shiitake mushroom and squid soup and a taste receptor. Through sensory evaluation and threshold determination, it is determined that the salty salt has remarkable salty taste and salty taste gaining effects under the low-sodium condition, the salty salt can replace part of edible salt to be applied to seasonings, a new choice of a natural flavoring agent is provided for low-salt food and seasonings, the requirement for healthy seasoning is met, and development of the food industry is promoted.

Description

Technical Field

[0001] The invention belongs to the technical field of protein active peptides, and particularly relates to a salty peptide derived from mushroom squid soup and an application thereof in food seasonings. Background Art

[0002] Salty peptides are compounds formed by two or more amino acids connected by peptide bonds. They are mainly extracted from food or directly synthesized from amino acids. Salty peptides have salty properties themselves, or they do not have salty properties themselves but can promote the release of salty taste and enhance the perception of salty taste. The emergence of salty peptides has created great value for the application of low-salt or salt-free foods. Salty peptides can effectively reduce the sodium content in food without reducing the original flavor of the food. This is of great significance for reducing sodium intake and preventing related health problems (such as hypertension) worldwide. For example, the combination of salty peptides with other seasonings such as NaCl and MSG can reduce the amount of sodium salt while maintaining the original taste of the food. The research on salty peptides has not only promoted the development of food science, but also provided new directions for innovation in the food industry. For example, the study of salty receptors and salty transduction mechanisms can help to understand human taste perception more deeply, thereby developing more foods that are beneficial to health. The development of salty peptides not only helps to improve the flavor and health value of food, but also promotes related scientific research and innovation in the food industry.

[0003] However, the existing salty peptides are still not widely used in low-salt foods, food industry, and pet food. With the improvement of consumers' health awareness, low-salt diet has gradually become a trend. Low-salt foods need to meet the demand for rich taste while maintaining health. Salty peptides have great potential in this field and can effectively enhance the taste of food, so that it can still provide a rich taste experience even when the salt content is reduced. The demand for salt in the food industry is mainly reflected in preservation, seasoning, and improving the taste of food. With the promotion of healthy diet, many food processing companies have begun to reduce the use of salt in food and look for suitable substitutes. Salty peptides can provide a substitute for salt without increasing sodium, which is an important direction for the future development of the food industry. In the field of pet food, with the increase of pet health problems, the ingredients of pet food are gradually developing towards low salt, low fat, high protein and other directions. As a low-sodium and efficient salty enhancer, salty peptides can enable pet food to provide palatability while reducing the potential impact on pet health.

[0004] Although there are many research results on salty peptides, more types of salty peptides need to be developed to meet the needs of low-salt foods, food industry, pet food and other fields. These new salty peptides should not only have good salty taste perception intensity, but also be optimized in production cost, stability, adaptability and other aspects to promote the popularization of healthier foods and eating patterns. Summary of the invention

[0005] In order to overcome the deficiencies of the above-mentioned prior art, the purpose of the present invention is to provide a salty peptide derived from mushroom squid soup and its application in food seasoning, which has the characteristics of significant saltiness enhancement effect, reducing the amount of edible salt in food, and having a wide range of flavor enhancement fields.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] The salty peptide derived from mushroom squid soup is characterized in that the salty peptide can bind to the salty receptor SCNN1A, and the amino acid sequence of the salty peptide is shown in SEQ ID NO.1, which is GMGQKDSYVGDEAQSKRGILTL.

[0008] The salty peptide derived from mushroom squid soup is used in preparing food seasoning.

[0009] The food is low-salt food. Low-salt food refers specifically to those foods with sodium content strictly controlled within a low range of no more than 120 mg per 100 grams of solid food or 100 ml of liquid food. It is designed to meet the nutritional needs of special groups such as infants, the elderly, and patients with cardiovascular diseases, as well as the healthy diet standards of pets, and is intended to effectively reduce the risk of excessive sodium intake, thereby maintaining the health of these specific groups.

[0010] The food is a flour product or a meat product.

[0011] The seasoning is a salty seasoning.

[0012] The food seasoning is a low-sodium food additive for compound seasoning powder, seasoning sauce and liquid seasoning.

[0013] The low-salt food is food for infants and the elderly, nutritional dietary food for cardiovascular patients or pet food.

[0014] A salt substitute seasoning, comprising, by mass percentage, 55% to 70% of a main seasoning, 25% to 35% of an auxiliary material and 1% to 6% of umami peptide, with the remainder being spices;

[0015] The main seasoning is selected from one or more of monosodium glutamate, yeast extract, oyster sauce, fish sauce, edible salt and disodium 5'-ribonucleotide, the auxiliary materials are selected from one or more of corn starch, xanthan gum, glucose, konjac flour, polyglutamic acid, soy protein, pea starch, potato starch, wheat starch, rice starch, tapioca starch, erythritol, red bean powder and calcium stearate, and the amino acid sequence of the salty peptide is shown in SEQ ID NO.1.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The salty peptide derived from mushroom squid soup provided by the present invention is obtained by protease hydrolysis and chromatographic separation technology. The obtained salty peptide is subjected to molecular docking and sensory evaluation, and it is found that it has a specific salty threshold, has a significant salty taste and salty taste gain effect under low sodium conditions, provides a salty taste similar to traditional edible salt at a lower concentration, can replace part of the edible salt and be applied to seasonings, reduce the use of edible salt, and help consumers reduce sodium intake, which is of great significance for controlling hypertension and preventing cardiovascular diseases. While reducing the use of edible salt, it can help maintain the flavor of food or beverages, make low-salt foods closer to full-salt foods in taste, improve consumer acceptance and food quality, make products more competitive in the market, meet the needs of healthy seasoning, and expand the application prospects of edible fungus flavor peptides in the field of food flavor enhancement. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is the mass spectrum result diagram of GMGQKDSYVGDEAQSKRGILTL of the present invention.

[0019] Figure 2 This is the docking diagram of GMGQKDSYVGDEAQSKRGILTL and SCNN1A of the present invention.

[0020] Figure 3 This is a surface force analysis diagram of the interaction between SCNN1A of the present invention and the GMGQKDSYVGDEAQSKRGILTL salty peptide.

[0021] Figure 4 The figure is a comparison chart of the salty taste effects of different edible salt concentrations and peptide solutions; among them, a~f represent significant difference analysis charts.

[0022] Figure 5 The diagram shows the saltiness enhancement effect of different salty peptide concentrations in salt substitute seasonings; among them, a~g represent significant difference analysis diagrams. DETAILED DESCRIPTION

[0023] The present invention is further described in detail below in conjunction with specific embodiments, which are intended to explain the present invention rather than to limit it.

[0024] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0025] It should be noted that the salty peptide derived from mushroom squid soup involved in this article is not limited to being extracted according to the extraction method given in the following examples, but can also be obtained by artificial synthesis according to the amino acid sequence of the salty peptide derived from mushroom squid soup given in the examples.

[0026] Example 1

[0027] The salty peptide derived from shiitake mushroom squid soup can bind to the salty receptor SCNN1A. The amino acid sequence of the salty peptide is shown in SEQ ID NO.1, which is GMGQKDSYVGDEAQSKRGILTL.

[0028] Example 2

[0029] Application of the salty peptide derived from mushroom squid soup in preparing food seasoning;

[0030] The food is a low-salt food;

[0031] The food is a flour product;

[0032] The seasoning is a salty seasoning;

[0033] The food seasoning is a compound seasoning powder;

[0034] The low-salt foods are foods for infants and the elderly. Low-salt foods specifically refer to foods with sodium content strictly controlled within a low range of no more than 120 mg per 100 grams of solid foods or 100 ml of liquid foods. They are designed to meet the nutritional needs of special groups such as infants, the elderly, and patients with cardiovascular diseases, as well as the healthy dietary standards for pets, and are designed to effectively reduce the risk of excessive sodium intake, thereby maintaining the health of these specific groups.

[0035] Example 3

[0036] Application of the salty peptide derived from mushroom squid soup in preparing food seasoning;

[0037] The food is a low-salt food;

[0038] The food is a meat product;

[0039] The seasoning is a salty seasoning;

[0040] The food seasoning is a seasoning sauce;

[0041] The low-salt food is a nutritious dietary food for cardiovascular patients.

[0042] Example 4

[0043] Application of the salty peptide derived from mushroom squid soup in preparing food seasoning;

[0044] The food is a low-salt food;

[0045] The food is a flour product;

[0046] The seasoning is a salty seasoning;

[0047] The food seasoning is a liquid seasoning;

[0048] The low-salt food is pet food.

[0049] Example 5

[0050] A salt substitute seasoning, comprising, by mass percentage, 55% of a main seasoning, 25% of an auxiliary material and 1% of a umami peptide, with the remainder being spices;

[0051] Wherein, the main seasoning is selected from sodium glutamate, the auxiliary material is selected from corn starch, and the amino acid sequence of the salty peptide is shown as SEQ ID NO.1.

[0052] Example 6

[0053] A salt substitute seasoning, comprising, by mass percentage, 63% of a main seasoning, 30% of an auxiliary material and 4% of a umami peptide, with the remainder being spices;

[0054] The main seasoning is selected from oyster sauce, the auxiliary material is selected from soybean protein, and the amino acid sequence of the salty peptide is shown as SEQ ID NO.1.

[0055] Example 7

[0056] A salt substitute seasoning, comprising, by mass percentage, 70% of a main seasoning, 35% of an auxiliary material and 6% of umami peptide, with the remainder being spices;

[0057] The main seasoning is selected from 5'-flavor ribonucleotide disodium, the auxiliary material is selected from red bean powder, and the amino acid sequence of the salty peptide is shown in SEQ ID NO.1.

[0058] Example 8 Extraction of salty peptides from mushroom squid soup

[0059] 1) Take 1.6 kg of pork shank, wash and cut into pieces, put into a pot with cold water, add 10 g of scallion, 10 g of ginger, and 10 g of cooking wine, and blanch for 30 minutes, skim off the foam, and then remove and wash to remove blood. After frying the pork shank for 10 minutes, add 4 L of water, 10 g of scallion, 10 g of ginger, 10 g of cooking wine, and 5 g of salt and cook for 4 hours to make the soup.

[0060] 2) Take 60 g of evenly matched squid rings, wash and blanch for 5 min, peel them, add 3 g of scallion, 3 g of ginger and 5 g of cooking wine, and then add the broth prepared in step 1 at a liquid-to-solid ratio of 2.5:1 to obtain squid soup.

[0061] 3) Add 9.6 g of shiitake mushrooms to the squid soup obtained in step 2, cook at 74°C for 1.5 h to obtain shiitake mushroom squid soup, and cool it down.

[0062] 4) Pour the mushroom squid soup obtained in step 3 into a culture dish, freeze it at -80°C for 3 h, and then take it out and put it into a vacuum freeze dryer for 2 days.

[0063] 5) Take an appropriate amount of the sample obtained in step 4 and desalt it through a C18 desalting column, and analyze the sample via LC-MS / MS equipped with an online nanospray ion source.

[0064] Example 9 Identification of salty peptides from mushroom squid soup

[0065] The peptide obtained in step 1 was sequenced by mass spectrometry. The mass spectrometry results were as follows: Figure 1 The amino acid sequence of the polypeptide is shown in SEQ ID NO. 1, specifically: Gly-Met-Gly-Gln-Lys-Asp-Ser-Tyr-Val-Gly-Asp-Glu-Ala-Gln-Ser-Lys-Arg-Gly-Ile-Leu-Thr-Leu, with a molecular weight of 2352.17 Da, a length of 22, and a molecular mass-to-charge ratio of 589.05.

[0066] Example 10 Evaluation of the Taste Properties of Salty Peptides from Mushroom Squid Soup

[0067] Since the amiloride-sensitive sodium ion channel encoded by SCNN1A plays an important role in taste perception, especially closely related to the perception of salty taste, it can respond to the stimulation of sodium ions and transmit signals to taste cells, thereby inducing nerve conduction and making us perceive salty taste. In this experiment, SCNN1A was used as a salty taste receptor, and the AlphaFold2 software was used to establish a model of the salty taste receptor SCNN1A and the peptide extracted in step 1. The pdb files of the salty taste receptor SCNN1A and the extracted peptide were selected, and molecular docking was performed through Discovery Studio 2019. The docking results are shown in the figure below. Figure 2 and Figure 3 shown.

[0068] Depend on Figure 2 and Figure 3It can be seen that the polypeptide with the amino acid sequence shown in SEQ ID NO.1 successfully docked with the salty taste receptor SCNN1A, and the binding sites were Tyr302, Met301, Thr345, Arg249, Glu250, Asp246, Ser242, Leu210, Ser209, Thr646, Thr638, Ala636, Pro635, Ser634, Pro633 and Ala632. Among them, Asp246, Arg249, Leu210, Thr646, Thr638, Ala636 and Ser634 interacted with the polypeptide with the amino acid sequence shown in SEQ ID NO.1 through hydrogen bonds, and Ser242, Thr345, Ala636 and Ser209 interacted with the polypeptide with the amino acid sequence shown in SEQ ID NO.1 through carbon-hydrogen bonds. The docking results showed that the sequences mainly interacted through hydrogen bonds and carbon-hydrogen bonds, indicating that the polypeptide extracted in Example 8 was a salty peptide.

[0069] Example 11 Sensory Evaluation of Salty Peptides from Mushroom Squid Soup

[0070] Ten volunteers (5 males and 5 females, aged between 23 and 26 years old) without taste disorders and no history of taste disorders were recruited and trained in advance to prepare for the basic taste of salty taste, namely NaCl. In order to avoid the influence of the previous sample on the next sample, the panelists were required to rinse their mouths with 50 mL of pure water between two samples. The sensory evaluation of taste peptides was carried out by quantitative descriptive analysis. First, the peptide was synthesized according to the amino acid sequence shown in SEQ ID NO.1, and then the synthesized peptide was fully dissolved in ultrapure water to prepare a peptide solution with a concentration of 2 mg / mL. The peptide solution was divided into 3 parts and then filled into sterilized bottles (each bottle was randomly numbered). 0.1% NaCl, 0.35% NaCl and 0.6% NaCl were added to the peptide solution as the test solution; at the same time, 3 different sterilized bottles were taken, and 0.1% NaCl, 0.35% NaCl and 0.6% NaCl were added into them as the control solution, and then the solutions in each group were tasted and scored. Ask the panelists to hold the sample in their mouth for 10 seconds, describe the taste of the sample, such as salty, and rate its intensity. After scoring, rinse the mouth with 50 mL of pure water and wait for at least 1 minute for the taste in the mouth to dissipate before tasting and scoring the next set of solutions. Finally, calculate the average value based on the scoring results.

[0071] The scoring results of each group are as follows Figure 4As shown, it can be seen that after adding 0.1% NaCl, 0.35% NaCl, and 0.6% NaCl to the polypeptide solution, the sensory score was significantly higher than that of 0.1% NaCl, 0.35% NaCl, and 0.6% NaCl solutions (P < 0.05), indicating that the flavor peptide can increase the saltiness intensity of NaCl to a certain extent, and can be used as a base material or auxiliary material in the field of food seasoning.

[0072] Example 12 Application of salty peptides derived from mushroom squid soup

[0073] A salt substitute seasoning, comprising 55% to 70% of a main seasoning, 25% to 35% of an auxiliary material and 1% to 6% of umami peptide, with the remainder being spices;

[0074] The main seasoning is selected from one or more of monosodium glutamate, yeast extract, oyster sauce, fish sauce, edible salt and disodium 5'-ribonucleotide, the auxiliary materials are selected from one or more of corn starch, xanthan gum, glucose, konjac flour, polyglutamic acid, soy protein, pea starch, potato starch, wheat starch, rice starch, tapioca starch, erythritol, red bean powder and calcium stearate, and the amino acid sequence of the salty peptide is shown in SEQ ID NO.1.

[0075] The production process of the above-mentioned salt substitute seasoning is as follows:

[0076] ① Crushing of raw and auxiliary materials: crush the main seasoning into powder using a grinder.

[0077] ② Stirring and mixing: weigh the auxiliary materials and spices and mix them for 3 to 5 minutes, then add a certain proportion of salty peptide mixture for about 6 to 10 minutes to mix all the additives evenly.

[0078] ③ Rotary granulation: Slowly pour the evenly mixed wet material into the trough. The wet material is pushed by the S-shaped milling knife and squeezed into shape using a sieve cylinder with an aperture of 1.2mm.

[0079] ④ Drying and sieving: Place the granulated product in a 60℃ oven and dry for 4 hours. After drying, sieve it through a 10-mesh sieve and then a 16-mesh sieve to collect particles between 10 and 16 mesh as the final product.

[0080] ⑤Packaging: After drying, immediately cool to around 30~45℃ and package.

[0081] According to the above production process, salt substitute seasonings with different salty peptide addition amounts were prepared to study the effect of the salty peptide addition amount on the salty effect. The steps are as follows:

[0082] The salt-substitute seasoning without added salty peptides includes, by mass percentage, 35% of monosodium glutamate, 25% of edible salt, 15% of corn starch, 20% of wheat starch and 5% of spices.

[0083] The salt substitute seasoning with 1% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 19% wheat starch, 1% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0084] The salt substitute seasoning with 2% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 18% wheat starch, 2% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0085] The salt substitute seasoning with 3% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 17% wheat starch, 3% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0086] The salt substitute seasoning with 4% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 16% wheat starch, 4% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0087] The salt substitute seasoning with 5% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 15% wheat starch, 5% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0088] The salt substitute seasoning with 6% salty peptide added includes, by mass percentage, 35% sodium glutamate, 25% edible salt, 15% corn starch, 14% wheat starch, 6% salty peptide as shown in SEQ ID NO.1 and 5% spices.

[0089] According to the sensory evaluation method of Example 11, the salt-replacing seasonings with different amounts of salty peptides added were scored.

[0090] The scoring results of each group are as follows Figure 5 As shown, as the content of salty peptides from mushroom squid soup increases, the salty effect of the salt substitute seasoning becomes stronger. Therefore, adding the above salt substitute seasoning to food can achieve a low-sodium flavoring effect while ensuring the salty taste, and is suitable for adding to infant and elderly food, nutritional diet for cardiovascular patients, and pet food.

[0091] The above contents are only for explaining the technical idea of ​​the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the claims of the present invention.

Claims

1. A salty peptide derived from mushroom squid soup, characterized in that: The salty peptide can bind to the salty taste receptor SCNN1A. The amino acid sequence of the salty peptide is shown in SEQ ID NO.1, which is GMGQKDSYVGDEAQSKRGILTL.

2. Use of the salty peptide derived from mushroom squid soup according to claim 1 in preparing food seasonings.

3. The use according to claim 2, characterized in that: The food is a low-salt food.

4. The use according to claim 2, characterized in that: The food is a flour product or a meat product.

5. The use according to claim 2, characterized in that: The seasoning is a salty seasoning.

6. The use according to claim 2, characterized in that: The food seasoning is a low-sodium food additive for compound seasoning powder, seasoning sauce and liquid seasoning.

7. The use according to claim 3, characterized in that: The low-salt food is food for infants and the elderly, nutritional dietary food for cardiovascular patients or pet food.

8. A salt substitute seasoning, characterized in that: In terms of mass percentage, it includes 55% to 70% of main seasonings, 25% to 35% of auxiliary materials and 1% to 6% of umami peptides, and the remainder is spices; The main seasoning is selected from one or more of monosodium glutamate, yeast extract, oyster sauce, fish sauce, edible salt and disodium 5'-ribonucleotide, the auxiliary materials are selected from one or more of corn starch, xanthan gum, glucose, konjac flour, polyglutamic acid, soy protein, pea starch, potato starch, wheat starch, rice starch, tapioca starch, erythritol, red bean powder and calcium stearate, and the amino acid sequence of the salty peptide is shown in SEQ ID NO.1.

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