CN110305192B - Delicate flavor polypeptide and flavor composition and application thereof - Google Patents

Abstract

The invention relates to a polypeptide, and particularly discloses an umami polypeptide, a flavor composition and an application thereof.

Description

Delicate flavor polypeptide and flavor composition and application thereof

Technical Field

The invention belongs to the field of food, and particularly relates to an umami polypeptide, a flavor composition and application thereof.

Background

The basic tastes of food include sour, sweet, bitter, salty and fresh, which are the tastes generated by stimulating taste buds on the surface of tongue by soluble components in food dissolved in saliva or food solution, transmitting to the taste center of brain through taste nerve fiber, and analyzing by brain. Umami is a complex combination of tastes and is the complex perception of human receptors on the tongue of the umami component in food. The addition of the flavor enhancer to the food can not only improve the overall taste intensity of the food, but also enhance the flavor characteristics of the food, such as persistence, thickness and the like.

The main flavoring agents commonly used for delicate flavor at present are amino acids, nucleotides, organic acids and compound flavoring agents. Wherein the amino acids include sodium glutamate, L-alanine, L-aspartic acid and L-glycine, the nucleotides include disodium 5 '-Inosinate (IMP) and disodium 5' -Guanylate (GMP), the organic acids include disodium succinate, and the compound flavoring agent mainly comprises sodium glutamate and the nucleotides to generate delicate flavor. In actual production, sodium glutamate is often added to improve the delicate flavor of meat-flavored essence or other foods. Compared with sodium glutamate, the flavoring polypeptide has natural taste of food, can maintain the original natural taste of food as flavoring agent, has good safety, and has certain nutritive value. At present, the application of the flavor-developing polypeptide in the aspect of food seasoning is more and more emphasized.

Peptides are known as effective taste components, and it is reported that sweet, salty, bitter, and umami peptides play an important role in the development of characteristic flavors of fermented products, pickled and smoked products, and impart a specific flavor to foods. The umami was first discovered and named by japanese scientist Ikeda in 1908. The umami taste of peptides has received increasing attention since the isolation of the umami peptide Lys-Gly-Asp-Glu-Glu-Ser-Leu-Ala from beef by Yamasaki et al in 1978. As the peptide having umami taste, acidic peptides having a molecular weight of 1000 or less obtained by enzymatically decomposing a fish protein are known, and peptides having an amino acid sequence such as Glu-Asp, Thr-Glu, Glu-Asp-Glu and the like are known (Journal of Agricultural & Food Chemistry,23,49-53,1975).

Patent 100525647 discloses a seasoning containing umami peptide, wherein the peptide as an active ingredient in the seasoning is a peptide consisting of amino acid sequences represented by Gly-Pro-3Hyp, Gly-Pro-4Hyp, and Gly-Pro-Ala. Patent document 108623660 discloses an umami tetradecapeptide with the chemical structure RGENESDEQGAIVT (wherein R is arginine, G is glycine, E is glutamic acid, N is asparagine, S is serine, D is aspartic acid, Q is glutamine, a is alanine, I is isoleucine, V is valine, T is threonine, all amino acids are in L configuration).

Since peptides have various effects on the taste of foods and beverages, even though more and more umami peptides are reported, it is desired to develop peptides that can effectively enhance the umami taste of foods and beverages.

The composition provided by the invention contains four polypeptides with delicate flavor, can be used in food and beverage independently or randomly in combination, and is used for improving the delicate flavor of the food and beverage.

Disclosure of Invention

The invention aims to provide a polypeptide capable of enhancing delicate flavor and a flavor composition thereof in combination with the defects of the prior art.

Compared with amino acid flavor enhancers, the flavor enhancer has softer and mellow flavor and good aftertaste, and has flavor synergistic effect when being mixed with other flavor enhancers.

It is another object of the present invention to provide a food product having enhanced umami taste using the above flavor composition.

The specific scheme of the invention is as follows:

an umami polypeptide, the structure of which is Val-His-Ala-His-Ser.

An umami polypeptide, the structure of which is Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala.

An umami polypeptide, the structure of which is Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly.

An umami polypeptide, the structure of which is Leu-Val-Gln-Tyr.

Ala represents alanine, Glu represents glutamic acid, Gln represents glutamine, Tyr represents tyrosine, Val represents valine, Lys represents lysine, His represents histidine, Arg represents arginine, Leu represents leucine, Asn represents asparagine, Pro represents proline, Gly represents glycine, and Ser represents arginine.

Preferably, the four polypeptides are isolated from hydrolyzed animal and/or vegetable proteins.

Such as hydrolyzed meat or soybean protein, and separating by ultrafiltration, gel chromatography, high performance liquid chromatography, etc. to obtain the peptide or its salt for enhancing umami taste.

Preferably, the amino acids in all four of the above polypeptides are in the L-configuration.

The invention also relates to a polypeptide salt, which is prepared from one or more of the polypeptides.

Preferably, the polypeptide salt is a sodium salt of the polypeptide.

The invention also relates to a flavour composition comprising one or more of the above polypeptides and/or a salt of the above polypeptides.

Preferably, the flavour composition comprises only Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala polypeptides or salts thereof for enhancing umami taste.

Preferably, the flavour composition comprises only Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly polypeptide or salt thereof for enhancing umami taste.

Preferably, only Val-His-Ala-His-Ser containing polypeptide or salt thereof is used in the flavour composition to enhance umami taste.

Preferably, only the Leu-Val-Gln-Tyr polypeptide or salt thereof is included in the flavor composition for enhancing umami taste.

Preferably, the flavor composition contains two or more polypeptides selected from Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala, Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly, Val-His-Ala-His-Ser, and Leu-Val-Gln-Tyr, or salts thereof.

Preferably, the polypeptides Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala, Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly, Val-His-Ala-His-Ser, Leu-Val-Gln-Tyr or salts thereof are present in a molar ratio of (0-10): (0-5).

Preferably, the polypeptides Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala, Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly, Val-His-Ala-His-Ser, Leu-Val-Gln-Tyr or salts thereof are present in a molar ratio of (0.5-10): (0.5-5).

The invention also relates to a food which contains an additive, wherein the additive is one or more of the polypeptides and/or the polypeptide salt and/or the flavor composition.

Preferably, the additive can be added directly to the food product in an amount of 0.000001 to 1.0% by weight of the food product.

Preferably, the additive may be added directly to the food product, the concentration of the additive in the food product being from 0.000001 to 0.005%.

The food may be a seasoning containing an additive. If the additive is blended with food comprising salt, the salt can be sodium chloride or potassium chloride, not only can improve the delicate flavor of the food, but also can reduce the using amount of the salt, and has the effect of improving the salty taste.

The flavoring agent can be liquid, granular or powder.

The food containing the additive of the present invention may further contain excipients such as organic acids, amino acids, saccharides, nucleic acids, and amylolytic dextrins.

The food to be the umami-enhancing food of the present invention may be any food. For example, soy sauce, solid sauce soup, sauce, dressing, salad dressing, or other sauce having a flavor, etc. It can also be clear soup, thallus Porphyrae soup, Undaria Pinnatifida soup, thallus Porphyrae soup, broth, egg soup, vegetable soup, etc. It can also be soup or seasoning juice of flour. It can also be made into cooking food such as porridge and dessert, processed food of livestock such as sausage, cheese and ham, pickled product, processed product of vegetables, snack such as potato chip and biscuit, etc.

The above-mentioned additive may be added to the food or drink to be prepared, or may be added to the food or drink after the preparation, a solution or a treated product of the solution containing the peptide or the salt thereof, or an additive containing the flavor composition.

The invention also relates to the application of the polypeptide or the polypeptide salt or the flavor composition in food.

The invention has the beneficial effects that:

the invention has the structure of polypeptide or salt thereof of Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala, Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly, Val-His-Ala-His-Ser, Leu-Val-Gln-Tyr, which is used as an effective component for improving the delicate flavor, can also show obvious delicate flavor feeling under low concentration and has good delicate flavor characteristic; when the additive is used for food or seasoning, the delicate flavor of the food or seasoning can be obviously improved.

Drawings

FIG. 1 is a secondary mass spectrum of positive ion detection of polypeptide Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala.

FIG. 2 is a secondary mass spectrum of positive ion detection of polypeptide Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly.

FIG. 3 is a secondary mass spectrum diagram of the positive ion detection of the polypeptide Val-His-Ala-His-Ser.

FIG. 4 is the secondary mass spectrum of the positive ion detection of the polypeptide Leu-Val-Gln-Tyr.

Detailed Description

The present invention will be described in further detail with reference to examples.

Example one

The polypeptides Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala (A), Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B), Val-His-Ala-His-Ser (C) and Leu-Val-Gln-Tyr (D) are prepared by solid phase synthesis.

Resin treatment: 0.45g of Fmoc-Rink amide-MBHA resin was weighed and placed in a reactor, the resin was swelled in 20mL of DCM for 30min, then the resin was washed with 7mL of DMF or DCM for 2 times (DMF and DCM washed with resin 1 times each, resin 1 time each) alternately, the solution was drained, then 7mL of deprotecting agent (20% piperidine/DMF solution) was added to the reactor and shaken at room temperature for 30min to remove the Fmoc protecting group from the resin, and then the resin was washed with 7mL of DMF or DCM alternately for 2 times, respectively, for further use.

(1) The synthesis of the polypeptide A comprises the following steps:

Fmoc-Ala (0.6mmol), activating agent (0.6mmol HBTU and 0.6mmol HOBt) and activating base (1.2mmol DIPEA) were dissolved in 7mL DMF and added to the reactor to mix with the above treated resin, reacted at room temperature for 1h, after which the reaction solution was removed by filtration, the resin was washed 3 times with 7mL DMF or DCM alternately (1 time each DMF and DCM), and the same procedure was repeated, adding 7mL deprotecting agent (20% piperidine/DMF solution) to deprotect, and then adding Fmoc-amino acid (0.6mmol), activating agent (0.6mmol HBTU and 0.6mmol HOBt) and activating base (1.2mmol DIPEA) to react with Fmoc-Gln, Fmoc-Asn, Fmoc-Ser, Fmoc-Tyr, Fmoc-Pro, Fmoc-His, Fmoc-Ala in order to link to Gln, Asn, Tyr, and activating base (1.2mmol DIPEA), respectively, Pro, His, Ala. Finally, the obtained polypeptide-resin compound is put into a reactor, 7mL of DCM or methanol is alternately cleaned for 3 times (methanol and DCM respectively wash the resin for 1 time, and the resin is washed alternately for 1 time), 10mL of cutting agent (95% trifluoroacetic acid solution) is added into the reactor, the shaking reaction is carried out for 30min at 0 ℃, the shaking reaction is carried out for 3h at room temperature, the liquid is filtered into 100-500mL of glacial ethyl ether, the frozen centrifugation and the vacuum drying are carried out, and then a Zorbax300SB-C18(9.4mm multiplied by 250mm, 5 mu m) chromatographic column is adopted for separation and purification, and the mobile phase comprises A water/trifluoroacetic acid (1: 0.001, v/v), B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 mu L of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the machine model is a liquid mass spectrometer 5800 MALDI-TOF mass spectrometer (AB Sciex company, USA), the obtained polypeptide can be confirmed by positive ion detection secondary mass spectrometry, 0.3g of polypeptide A is obtained by freeze-drying, and the spectrogram of the positive ion detection secondary mass spectrometry of the polypeptide A is shown in figure 1.

(2) The synthesis of the polypeptide B comprises the following steps:

Fmoc-Ala (0.6mmol), activating agent (0.6mmol HBTU and 0.6mmol HOBt) and activating base (1.2mmol DIPEA) are dissolved in 7mL DMF and added into a reactor to be mixed with the treated resin, the mixture is reacted for 1h at room temperature, after the reaction is finished, the reaction solution is filtered and removed, the resin is washed 3 times by 7mL DMF or DCM alternately (the resin is washed 1 time by DMF and DCM respectively), and the reaction is carried out sequentially with Fmoc-Glu, Fmoc-Gln, Fmoc-Tyr, Fmoc-Arg, Fmoc-Leu, Fmoc-Val and Fmoc-Gly by the same polypeptide A synthesis method so as to connect Glu, Gln, Tyr, Arg, Leu, Val and Gly. Finally, the obtained polypeptide-resin compound is put into a reactor, 7mL of DCM or methanol is alternately cleaned for 3 times (methanol and DCM respectively wash the resin for 1 time, and the resin is washed alternately for 1 time), 10mL of cutting agent (95% trifluoroacetic acid solution) is added into the reactor, the shaking reaction is carried out for 30min at 0 ℃, the shaking reaction is carried out for 3h at room temperature, the liquid is filtered into 100-500mL of glacial ethyl ether, the frozen centrifugation and the vacuum drying are carried out, and then a Zorbax300SB-C18(9.4mm multiplied by 250mm, 5 mu m) chromatographic column is adopted for separation and purification, and mobile phase components A water/trifluoroacetic acid (1: 0.001, v/v) and B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 mu L of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the machine model is a liquid mass spectrometer 5800 MALDI-TOF mass spectrometer (AB Sciex company, USA), the obtained polypeptide can be confirmed by positive ion detection secondary mass spectrometry, 0.25g of polypeptide B is obtained by freeze-drying, and the spectrum of the positive ion detection secondary mass spectrometry of the polypeptide B is shown in figure 2.

(3) The synthesis of the polypeptide C comprises the following steps:

dissolving Fomc-Val (0.6mmol), an activating agent (0.6mmol HBTU and 0.6mmol HOBt) and an activating base (1.2mmol DIPEA) in 7mLDMF, adding the solution into a reactor, mixing with the treated resin, reacting at room temperature for 1h, filtering to remove reaction liquid after the reaction is finished, washing the resin for 3 times by 7mLDMF or DCM (washing the resin for 1 time by DMF and DCM respectively, and washing the resin for 1 time alternately), and reacting with Fmoc-His, Fmoc-Ala, Fmoc-His and Fmoc-Ser in sequence by the same polypeptide A synthesis method to connect His, Ala, His and Ser. Finally, the obtained polypeptide-resin compound is put into a reactor, 7mL of DCM or methanol is alternately washed for 3 times (the resin is washed by methanol and DCM for 1 time, and the resin is washed by methanol and DCM for 1 time), 10mL of cutting agent (95% trifluoroacetic acid solution) is added into the reactor, the shaking reaction is carried out for 30min at 0 ℃, the shaking reaction is carried out for 3h at room temperature, the liquid is filtered into 100-500mL of glacial ethyl ether, the frozen centrifugation and the vacuum drying are carried out, and then a Zorbax300SB-C18(9.4mm multiplied by 250mm, 5 mu m) chromatographic column is adopted for separation and purification, and mobile phase compositions A water/trifluoroacetic acid (1: 0.001, v/v), B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 muL of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the machine model is a liquid mass spectrometer 5800 MALDI-TOF mass spectrometer (AB Sciex company, USA), the polypeptide can be confirmed by positive ion detection secondary mass spectrometry, 0.3g of polypeptide C is obtained by freeze-drying, and the spectrogram of the positive ion detection secondary mass spectrometry of the polypeptide C is shown in figure 3.

(4) The synthesis of the polypeptide D comprises the following steps:

Fmoc-Leu (0.6mmol), an activating agent (0.6mmol HBTU and 0.6mmol HOBt) and an activating base (1.2mmol DIPEA) are dissolved in 7mL DMF and added into a reactor to be mixed with the treated resin, the reaction is carried out for 1h at room temperature, after the reaction is finished, the reaction solution is filtered and removed, the resin is washed by 7mL DMF or DCM for 3 times alternately (the resin is washed by DMF and DCM for 1 time respectively, and the resin is washed by 1 time alternately), and the reaction is carried out with Fmoc-Val, Fmoc-Gln and Fmoc-Tyr sequentially by the same polypeptide A synthesis method to connect Val, Gln and Tyr. Finally, the obtained polypeptide-resin compound is put into a reactor, 7mL of DCM or methanol is alternately washed for 3 times (the resin is washed by methanol and DCM for 1 time, and the resin is washed by methanol and DCM for 1 time), 10mL of cutting agent (95% trifluoroacetic acid solution) is added into the reactor, the shaking reaction is carried out for 30min at 0 ℃, the shaking reaction is carried out for 3h at room temperature, the liquid is filtered into 100-500mL of glacial ethyl ether, the frozen centrifugation and the vacuum drying are carried out, and then a Zorbax300SB-C18(9.4mm multiplied by 250mm, 5 mu m) chromatographic column is adopted for separation and purification, and mobile phase compositions A water/trifluoroacetic acid (1: 0.001, v/v), B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 muL of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the machine model is a liquid mass spectrometer 5800 MALDI-TOF mass spectrometer (AB Sciex company, USA), the polypeptide can be confirmed by positive ion detection secondary mass spectrometry, 0.5g of polypeptide D is obtained by freeze-drying, and the spectrum of the positive ion detection secondary mass spectrometry of the polypeptide D is shown in figure 4.

Example two

(1) Preparation of the polypeptide Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala (A), Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B), Val-His-Ala-His-Ser (C), Leu-Val-Gln-Tyr (D) by hydrolysis of animal proteins

The method comprises the following steps: 300g of chicken meat broth, defatting with petroleum ether for 2 times (300 mL each), and filtering the aqueous layer through a 0.45 μm filter membrane. And (3) carrying out Sephadex G-15 gel chromatography separation, wherein the sample loading is 5mL, the sample concentration is 100mg/mL, the ultra-pure water is eluted, the flow rate is 3.0mL/min, and the ultraviolet detection wavelength is 220 nm. Collecting fractions according to the peak, lyophilizing, and separating and purifying with Zorbax300SB-C18(9.4mm × 250mm, 5 μm) chromatographic column to obtain mobile phase composition A water/trifluoroacetic acid (1: 0.001, v/v), and B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 mu L of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the model of the machine is a liquid chromatograph-5800 MALDI-TOF mass spectrometer (AB Sciex company in America), the secondary mass spectrometry of positive ion detection is carried out, the obtained polypeptide can be confirmed, and Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B) is obtained by freeze-drying, Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala (A) is 10mg, Val-His-Ser (C) is 15mg, and Leu-Val-Gln-Tyr (D) is 15 mg.

(2) Preparation of the polypeptide Val-His-Ala-His-Ser (C), Leu-Val-Gln-Tyr (D) by hydrolysis of vegetable proteins

The method comprises the following steps: 100g of soy protein isolate was mixed with water at a ratio of 1:5(w/w) and hydrolyzed with 0.5% w/w (based on the weight fraction of the aqueous soy protein isolate) of trypsin at 55 ℃ for 1.5 h. And (3) performing Sephadex G-25 gel chromatography separation, wherein the sample loading is 10mL, the sample concentration is 200mg/mL, the ultra-pure water is eluted, the flow rate is 2.0mL/min, and the ultraviolet detection wavelength is 220 nm. Collecting fractions according to the peak, lyophilizing, and separating and purifying with Zorbax300SB-C18(9.4mm × 250mm, 5 μm) chromatographic column to obtain mobile phase composition A water/trifluoroacetic acid (1: 0.001, v/v), and B acetonitrile/trifluoroacetic acid (1: 0.001, v/v); the flow rate is 3 ml/min; gradient elution, 0-14min, 3 → 23% B; 14-16min, 23 → 75% B; 16-28min, 75 → 95% B. The column temperature is 30 ℃; the detection wavelength is 220nm, the sample concentration is 30mg/mL, 50 mu L of sample is injected each time, fractions are collected according to spectral peaks, fraction analysis is carried out by liquid chromatography and mass spectrometry, the model of a machine is a liquid chromatograph-5800 MALDI-TOF mass spectrometer (AB Sciex company in America), the secondary mass spectrometry of positive ion detection is carried out, the obtained polypeptide can be confirmed, and Val-His-Ala-His-Ser (C)15mg and Leu-Val-Gln-Tyr (D)15mg are obtained by freeze-drying.

EXAMPLE III

Taking polypeptide A as an example, 0.1g of polypeptide A prepared in the first example is dissolved in 10mL of water, added with a proper amount of sodium hydroxide to neutralize to pH7.5, and freeze-dried to remove water to obtain the sodium salt of polypeptide A.

Test example 1

The flavor composition comprising the polypeptides Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala (A), Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B), Val-His-Ala-His-Ser (C), Leu-Val-Gln-Tyr (D) was assayed for the profile of the amount used.

Sensory evaluation: 10 experienced evaluators trained in appearance, flavor and texture evaluations were picked for comparative evaluations. The total score is 10 points, wherein 0 is tasteless, 1 is umami/salty, 2 is umami/salty, 3 is umami/salty, 4 is umami/salty, 5 is umami/salty, 6 is umami/salty, 7 is umami/salty, 8 is umami/salty, 9 is umami/salty, and 10 is umami/salty. This rating includes the ability to use 0.1 points and thus has the potential of 100 rating differences, which can be extended beyond 10 points to include extreme ratings, if desired.

The polypeptides a-D in the flavour compositions were prepared as test samples of 10ppb, 100ppb, 1ppm and 10ppm water, and the oral profile data for this amount of product was collected and the results are shown in table 1.

Salty refers to a taste sensation associated with sodium chloride on the tongue; umami refers to the taste associated with monosodium glutamate on the tongue.

TABLE 1 results of the perception of the Properties in the mouth of the Polypeptides A-D in the flavour compositions

Test example 2

Use of a flavour composition in food products and condiments, the flavour composition being tested for its ability to enhance umami taste.

Adding 1mL of prepared polypeptide Val-His-Ala-His-Ser (C)10mg/mL of aqueous solution into 99mL of monosodium glutamate solution (0.05%) as a sample, and adding 1mL of distilled water into 99mL of monosodium glutamate solution (0.05%) as a control group.

② adding 1mL of prepared polypeptide (sodium salt of polypeptide C, 10mg/mL) aqueous solution into 99mL of monosodium glutamate solution (0.05%) as a sample, and adding 1mL of distilled water into 99mL of monosodium glutamate solution (0.05%) as a control group.

③ 1mL of the prepared aqueous solution of the polypeptide (5 mg/mL of the polypeptide A and 5mg/mL of the sodium salt of the polypeptide A) was added to 99mL of the monosodium glutamate solution (0.05%) to serve as a sample, and 1mL of distilled water was added to 99mL of the monosodium glutamate solution (0.05%) to serve as a control group.

Fourthly, diluting 2mg of polypeptide Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B) with 500mL of distilled water. 2mL of the diluted solution was added to 50mL of soy sauce as a sample, and raw soy sauce was used as a control.

Fifthly, weighing a mixture of the polypeptides Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly (B) and Val-His-Ala-His-Ser (C) according to the molar ratio of 2:1, and diluting the mixture with 500mL of distilled water. 2mL of the diluted solution was added to 50mL of soy sauce as a sample, and raw soy sauce was used as a control.

Sixthly, weighing a mixture of polypeptides Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala (A), Val-His-Ala-His-Ser (C) and Leu-Val-Gln-Tyr (D) according to a molar ratio of 5:2:3, and diluting the mixture with 100mL of distilled water. 1mL of the dilution was added to 100mL of the laver soup, and the seasoned laver soup was used as a sample. The original laver soup was used as a control group.

Seventhly, 2mg of the mixture of the polypeptides A, B, C and D is weighed according to the molar ratio of 1:1:1:1 and diluted with 100mL of distilled water. 1mL of the dilution was added to 100mL of the laver soup, and the seasoned laver soup was used as a sample. The original laver soup was used as a control group.

Eighthly, weighing 1mg of a mixture of the polypeptide A, B, C and the polypeptide D according to the molar ratio of 6:8:3:4, diluting the mixture with 100mL of distilled water to obtain a diluent, and adding 1mL of the prepared diluent into 49mL of chicken broth to serve as a sample; 1mL of distilled water was added to 49mL of chicken broth as a control.

Ninthly, weighing 1mg of a mixture of the polypeptides A, B, C and D according to a molar ratio of 6:8:3:4, diluting the mixture with 100mL of distilled water to obtain a diluent, and adding 1mL of the prepared diluent into 49mL of chicken broth to serve as a sample; 1mg of polypeptide A was diluted with 100mL of distilled water to obtain a diluted solution, and 1mL of the prepared diluted solution was added to 49mL of chicken broth as a control.

R2 mg of polypeptide C was diluted with 500mL of distilled water. 2mL of the diluted solution was added to 50mL of soy sauce as a sample, and raw soy sauce was used as a control.

10 experienced evaluators trained in appearance, flavor and texture evaluations were picked for comparative evaluations. The total score is 10, wherein 0 is none, 10 is very strong, and the specific score can be determined according to the above criteria. This rating includes the ability to use 0.1 points and thus has the potential of 100 rating differences, which can be extended beyond 10 points to include extreme ratings if desired.

The scoring results for the above applications are shown in table 2:

TABLE 2 results of evaluation of the use of flavor compositions in foods and seasonings

Name (R)	Control group (fraction)	Experimental group samples (minutes)
			Gourmet powder + sample (polypeptide C)	5	10
② monosodium glutamate + sample (sodium salt of polypeptide C)	5	9.8
			③ monosodium glutamate + sample (polypeptide A + sodium salt of polypeptide A)	5	9.9
Sauce + sample (polypeptide B)	2	8.5
			Soy sauce + sample (polypeptide B + C)	2	8.8
Sixth, Laver soup + sample (polypeptide A + C + D)	3	8.8
			Seventhly, the laver soup and the sample (polypeptide A + B + C + D)	3	9.0
Eight chicken soup + sample (polypeptide A + B + C + D)	2	9.0
			Ninthly Chicken soup + sample (polypeptide A + B + C + D)	8.5	9.0
Sample of soy sauce (polypeptide C)	2	8.6

As can be seen from table 1, the four polypeptides in the flavour composition of the invention show a pronounced umami perception at low concentrations, with good umami properties.

As can be seen from Table 2, the polypeptide or the salt thereof in the flavor composition can remarkably improve the delicate flavor intensity of monosodium glutamate, soy sauce, laver soup and chicken soup, and the delicate flavor score is greatly improved compared with that of a control group and even can reach 10 points, so that the flavor composition has a good application prospect in foods and seasonings.

The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.

Sequence listing

<110> Beijing university of Industrial and commercial

<120> delicate flavor polypeptide, flavor composition and application thereof

<130> 20190717

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 5

<212> PRT

<213> Artificial sequence

<400> 1

Val His Ala His Ser

1 5

<210> 2

<211> 4

<212> PRT

<213> Artificial sequence

<400> 2

Leu Val Gln Tyr

1

<210> 3

<211> 8

<212> PRT

<213> Artificial sequence

<400> 3

Ala Gln Asn Ser Tyr Pro His Ala

1 5

<210> 4

<211> 8

<212> PRT

<213> Artificial sequence

<400> 4

Ala Glu Gln Tyr Arg Leu Val Gly

1 5

Claims (9)

1. The umami polypeptide is characterized in that the amino acid sequence of the polypeptide is Val-His-Ala-His-Ser.

2. The polypeptide of claim 1, wherein all of the amino acids in the polypeptide are in the L-configuration.

3. A salt of the polypeptide of claim 1 or 2.

4. A flavour composition comprising a polypeptide according to any one of claims 1-2 and/or a salt of a polypeptide according to claim 3.

5. Flavour composition according to claim 4, characterized in that said composition further comprises at least one polypeptide of Ala-Gln-Asn-Ser-Tyr-Pro-His-Ala, Ala-Glu-Gln-Tyr-Arg-Leu-Val-Gly, Leu-Val-Gln-Tyr or a salt thereof.

6. A food product comprising an additive which is a polypeptide according to any one of claims 1 to 2 and/or a salt of a polypeptide according to claim 3 and/or a flavour composition according to any one of claims 4 to 5.

7. The food product of claim 6, wherein the additive is added in an amount of 0.000001 to 1.0% by weight of the food product.

8. The food product of claim 7, wherein the additive is present in the food product at a concentration of 0.000001 to 0.005%.

9. Use of a polypeptide according to any of claims 1-2 or a salt of a polypeptide according to claim 3 or a flavour composition according to any of claims 4-5 as an additive in a food product.

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