CN108623660B

CN108623660B - Delicious tetradecapeptide, Maillard reaction product and application thereof

Info

Publication number: CN108623660B
Application number: CN201810443024.2A
Authority: CN
Inventors: 苏国万; 赵谋明; 张佳男
Original assignee: South China University of Technology SCUT
Current assignee: South China University of Technology SCUT
Priority date: 2018-05-10
Filing date: 2018-05-10
Publication date: 2021-05-14
Anticipated expiration: 2038-05-10
Also published as: CN108623660A

Abstract

The invention discloses an umami tetradecapeptide and a Maillard reaction product thereof, wherein the primary structure amino acid sequence of the polypeptide is RGENESDEQGAIVT; the preparation method of the Maillard reaction product comprises the following steps: mixing the polypeptide with water and reducing sugar, adjusting the pH value of the reaction system to 5.0-6.5, and reacting at 80-110 ℃ for 30-240min to obtain the polypeptide. The invention discovers a polypeptide (tetradecapeptide) with outstanding delicate flavor, and the delicate flavor intensity and the freshness enhancing function of the polypeptide can be further improved by adopting the Maillard reaction technology; and the umami tetradecapeptide and the Maillard reaction product thereof are applied to food and seasonings, so that the umami and fullness of the food and the seasonings can be obviously improved.

Description

Delicious tetradecapeptide, Maillard reaction product and application thereof

Technical Field

The invention belongs to the field of food additives, and particularly relates to an umami tetradecapeptide, a Maillard reaction product thereof and application of the umami tetradecapeptide and the Maillard reaction product.

Background

Umami (umami) is similar to the feeling caused by sodium glutamate (MSG), is the fifth basic taste after sourness, sweetness, saltiness and bitterness, and has the characteristics of increasing the richness of food mouthfeel, enabling people to feel pleasure and the like. Umami was first identified by the Japanese chemist Ikeda in the beginning of the 20 th century as a taste that distinguished from the four basic tastes in seaweed soup, but was not accepted by Western scholars for a long time. Until 2002, scholars found umami receptors, umami was not identified as the fifth basic taste. In addition to the umami taste of sodium glutamate and flavor nucleotides (IMP, GMP), researchers also find small molecule peptide substances with umami taste in hydrolyzed animal and plant proteins (HAP, HVP).

The fresh taste peptide is a peptide substance which can make up or enhance the original taste of food, can improve the quality of the food, improve the perception of taste and enhance the appetite of consumers. The production of the concept of the umami peptide comes from the delicious peptide (BMP), the delicious peptide is a peptide segment obtained by separating enzymolysis products of beef hydrolyzed by papain, and the primary structure of the delicious peptide is Lys-Gly-Asp-Glu-Ser-Leu-Leu-Ala after being identified. With the proposal of the delicious peptide, a great deal of research is carried out on the delicious peptide at home and abroad, and a plurality of delicious peptides are separated and identified from the proteolysis products.

Disclosure of Invention

The primary object of the present invention is to provide a tetradecapeptide with enhanced umami properties.

It is another object of the present invention to provide Maillard reaction products of the above polypeptides (tetradecapeptides) having more excellent flavor enhancing properties.

The invention further aims to provide the application of the tetradecapeptide and the Maillard reaction product thereof as a freshness-enhancing food additive.

The purpose of the invention is realized by the following technical scheme:

a polypeptide (tetradecapeptide) having a primary structure amino acid sequence of 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 the L configuration.

The above polypeptides may be produced by solid phase synthesis methods, coupling the amino acid sequences one by one on a resin to form the umami tetradecapeptide of the invention. For example, the synthesis is carried out on a solid phase synthesizer, 1g of dichloro resin is swelled and washed, Fmoc protecting groups are removed, amino acid is added for condensation reaction, and the deprotection-condensation process is repeated until all amino acids are connected.

The polypeptide can also be purified by adopting a reversed-phase high performance liquid chromatography, namely, firstly, the resin of the crude umami tetradecapeptide is cut, and then the crude umami tetradecapeptide is separated and purified on the reversed-phase high performance liquid chromatography to obtain the high-purity umami tetradecapeptide.

The Maillard reaction product of the polypeptide has more excellent freshness-increasing property, and is prepared by the following method:

mixing the polypeptide with water and reducing sugar, and heating to obtain Maillard reaction product of the polypeptide;

the heating is preferably carried out for reaction at the temperature of 80-110 ℃ for 30-240 min; the pH value of the reaction system is preferably 5.0-6.5;

in the reaction system of the method, the solid content is 40-60% (W/V), wherein the polypeptide content is 30-55% (W/V), and the reducing sugar content is 5-10% (W/V);

the reducing sugar is glucose and/or xylose.

The Maillard reaction has a very important influence on the color and flavor of food. In the presence of reducing sugars, the product after heat treatment gives a more pronounced taste than itself (polypeptide).

The polypeptide (tetradecapeptide) and the Maillard reaction product thereof can be used as a freshness-enhancing food additive for improving the delicate flavor and fullness of food.

Compared with the prior art, the invention has the following advantages and effects:

the invention discovers a polypeptide (tetradecapeptide) with outstanding delicate flavor, and the delicate flavor intensity and the function enhancement of the polypeptide can be further improved by adopting the Maillard reaction technology; and the umami tetradecapeptide and the Maillard reaction product thereof are applied to food and seasonings, so that the umami and fullness of the food and the seasonings can be obviously improved.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

In the following examples, the evaluation methods of the respective sensory evaluation were as follows:

(1) delicate flavour threshold value determination method

And performing sensory evaluation on the umami peptide by adopting a taste dilution analysis method. The umami peptide is accurately dissolved in purified water to prepare 1% concentration, then 1:1 (volume ratio) gradual dilution is carried out, the gradually diluted solution of the sample is presented to trained evaluators according to the concentration increasing sequence, and each dilution level solution is evaluated by adopting 3-point measurement. When the difference in flavor between a certain dilution level of the solution and 2 blanks (purified water) is just recognizable, the sample concentration at that time is said to be the recognizable threshold. The threshold value of each evaluation group is the average value of the evaluation results of each evaluator.

(2) Freshness-increasing threshold value determination method

And performing sensory evaluation on the umami peptide by adopting a comparative taste dilution analysis method. Changing blank (purified water) in taste dilution analysis method to 0.03mg/L monosodium glutamate solution, and performing other steps such as delicate flavor threshold determination method.

(3) Application of delicious peptide in food and condiment

Preparing the polypeptide prepared in each example into a solution of 10 mg/ml;

adding 1mL of distilled water into 99mL of monosodium glutamate solution (0.05%) as a control group, and adding 1mL of prepared polypeptide into 99mL of monosodium glutamate solution (0.05%) as a sample;

adding 1mL of distilled water into 99mL of soy sauce as a control group, and adding 1mL of prepared polypeptide into 99mL of soy sauce as a sample;

③ 1mL of distilled water was added to 49mL of chicken broth as a control, and 1mL of the prepared polypeptide was added to 49mL of chicken broth as a sample.

The sensory panel consisted of 10 panelists (five males and five females, aged 25 to 35 years) who were familiar with five basic taste qualities. Sensory evaluation the temperature in the room was controlled at 23. + -. 2 ℃. The samples were evaluated by scoring from 0 to 10, with 0 indicating no taste and 10 indicating a marked taste. The control group was selected as a standard with an umami score of 5, and different samples were scored on the basis of these standards, to finally form tables 1 and 2.

Example 1

A method for synthesizing polypeptide RGENESDEQGAIVT, comprising the steps of:

1g of dichloro resin is swelled and washed, Fmoc protecting groups are removed, amino acid is added for condensation reaction, and the deprotection-condensation process is repeated until all amino acids are connected. Cutting resin to obtain crude polypeptide RGENESDEQGAIVT, purifying by reverse phase high performance liquid chromatography, and freeze drying to obtain umami tetradecapeptide.

The umami taste and umami taste enhancing thresholds of the umami tetradecapeptide are shown in table 1.

The effect of the use of umami tetradecapeptide in a dressing is shown in table 2.

Example 2

A method for preparing a maillard reaction product of polypeptide RGENESDEQGAIVT, comprising the steps of:

mixing 3g of umami tetradecapeptide (RGENESDEQGAIVT), 1g of glucose and 6g of deionized water, adjusting the pH value of the system to 6.5 by adopting 0.1mol/L sodium hydroxide solution, reacting at 110 ℃ for 60min, cooling to normal temperature after the reaction is finished, and freeze-drying to obtain the umami tetradecapeptide Maillard reaction product 1.

The umami taste and freshness enhancement thresholds of the umami tetradecapeptide maillard reaction product 1 are shown in table 1.

The effect of the use of the umami tetradecapeptide maillard reaction product 1 in a condiment is shown in table 2.

Example 3

5.5g of umami tetradecapeptide (RGENESDEQGAIVT), 0.5g of glucose and 4g of deionized water are mixed, the pH value of the system is adjusted to 5.0 by adopting 0.1mol/L sodium hydroxide solution and 0.1mol/L HCL solution, the reaction is carried out for 20min at the temperature of 80 ℃, the temperature is reduced to normal temperature after the reaction is finished, and the product 2 of the Maillard reaction of the umami tetradecapeptide is obtained after freeze drying.

The umami taste and freshness enhancement thresholds of the umami tetradecapeptide maillard reaction product 2 are shown in table 1.

The effect of the use of the umami tetradecapeptide maillard reaction product 2 in a condiment is shown in table 2.

Example 4

4g of umami tetradecapeptide (RGENESDEQGAIVT), 0.8g of glucose, 0.2g of xylose and 6g of deionized water are mixed, the pH value of the system is adjusted to 6.0 by adopting 0.1mol/L sodium hydroxide solution, the reaction is carried out for 45min at 100 ℃, the temperature is reduced to normal temperature after the reaction is finished, and the product is frozen and dried to obtain the umami tetradecapeptide Maillard reaction product 3.

The umami taste and freshness enhancement thresholds of the umami tetradecapeptide maillard reaction product 3 are shown in table 1.

The effect of the use of the umami tetradecapeptide maillard reaction product 3 in a condiment is shown in table 2.

TABLE 1 Umamizymetrium and Umamizymetrium umami and umami enhancing thresholds of Maillard reaction products thereof

TABLE 2 Umamizymetrium and the use of Maillard reaction products thereof in seasonings

As can be seen from table 1, the umami threshold of the present invention, umami tetradecapeptide (example 1), was 0.43mmol/L, i.e., the umami tetradecapeptide showed a pronounced umami sensation at a concentration of about 0.075%, which was only 1.87 times the umami threshold of monosodium glutamate, and in addition, the umami tetradecapeptide showed a pronounced effect at a concentration of 0.38 mmol/L. Moreover, the Maillard reaction (examples 2 to 4) is performed on the umami tetradecapeptide, so that the umami intensity (the umami threshold value is 0.35 to 0.36mmol/L) and the umami effect (the umami threshold value is 0.21 to 0.23mmol/L) can be obviously improved. Therefore, the umami tetradecapeptide disclosed by the invention has a better umami characteristic, and the umami intensity of the umami tetradecapeptide can be obviously improved through a Maillard reaction.

As can be seen from table 2, the umami tetradecapeptide (example 1) and the maillard reaction products thereof (examples 2 to 4) of the present invention can significantly improve the umami strengths of monosodium glutamate, soy sauce and chicken broth, and the umami scores of the samples (monosodium glutamate, soy sauce and chicken broth) are increased from 5 to more than 7, even 9 (chicken broth + example 4). The invention shows that the delicious tetradecapeptide and the Maillard reaction product thereof have better application prospect in food and seasoning.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Sequence listing

<110> university of southern China's science

<120> umami tetradecapeptide, Maillard reaction product and application thereof

<140> 2018104430242

<141> 2018-05-10

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Arg Gly Glu Asn Glu Ser Asp Glu Gln Gly Ala Ile Val Thr

1 5 10

Claims

1. A polypeptide characterized by a primary structure amino acid sequence of RGENESDEQGAIVT.

2. A method for preparing a maillard reaction product of a polypeptide, comprising the steps of:

mixing the polypeptide of claim 1 with water and a reducing sugar and heating to obtain a maillard reaction product of the polypeptide;

the reducing sugar is glucose and/or xylose;

the heating is carried out for 30-240min at the temperature of 80-110 ℃.

3. The method of claim 2, wherein: in the reaction system, the solid content is 40-60% (W/V), wherein the polypeptide content is 30-55% (W/V), and the reducing sugar content is 5-10% (W/V).

4. A maillard reaction product of a polypeptide, characterized in that: is prepared by the method of claim 2 or 3.

5. Use of the polypeptide of claim 1 or the maillard reaction product of the polypeptide of claim 4 as a savory food additive.