JP5354520B2 - Novel peptide composition having kokumi imparting function - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition which can sustain and reinforce the tastes of foods, to provide a method for producing the composition, and to provide a food raw material containing the same. <P>SOLUTION: There is provided the composition as a seasoning ingredient, especially comprising a nucleic acid-based taste ingredient and one or more tetrapeptides represented by Leu-Glu-His-Ala, Ile-Lys-Gln-Asp, Ala-Gln-Pro-Thr, and Ala-Gln-Pro-Ala. The tetrapeptides can be used to reinforce the taste of a food and increase its body more than when the nucleic acid-based taste ingredient is used alone. <P>COPYRIGHT: (C)2010,JPO&amp;INPIT

Description

The present invention relates to a novel composition that can be added to foods to enhance the flavor and impart richness, fat feeling, richness, milky feeling, and the like, and an invention related thereto.

Peptides have not been well reported for their own taste, but are known to have a synergistic effect with other taste components. Typical examples include 1, umami 2, acidity 3, and salt.

1 is known to have a synergistic effect with umami components sodium inosinate (abbreviated as IMP), sodium guanylate (abbreviated as GMP), and amino acids, and to bring out umami. Non-Patent Document 1 reports that a peptide derived from chicken protein has a thick and delicious umami in the presence of IMP. Non-patent document 2 describes a cheese-derived peptide having a molecular weight of 1000 or less.
It has been reported that a water-soluble peptide having 4 to 5 residues contributes to the richness of cheese. Non-Patent Document 3 reports that a peptide having a molecular weight of 500 to 1000 derived from wheat gluten enhances the umami of boiled soup stock.

Regarding 2, the cooked pork protein-derived peptide has been reported to contribute to acidity suppression and saltiness enhancement (Non-patent Document 4).

Regarding 3, the fish soy sauce-derived peptide has been reported to enhance saltiness (Non-patent Document 5).

It has been reported that peptides contribute to taste and richness and thickness, and a taste-imparting peptide has been identified as a taste-contributing substance contained in yeast extract (Non-patent Document 6). The umami enhancement effect of peptides has been reported for peptides contained in livestock meat and plants. Patent Document 1 reports that a peptide derived from yeast extract enhances umami and has an excellent body taste and richness-imparting effect in foods and drinks, but the composition and structure of the peptide have been clarified. It wasn't. Thus, development of a useful new composition that maintains umami and imparts richness and thickness has been desired.

Biosci. Biotechnol. Biochem, Vol. 63 No. 3 p555-559, 1999 J. Daily Sci, Vol.90 No.11 p4966 ~ 4973, 2007 Biosci. Biotechnol. Biochem, Vol.68 No.8 p1657-1662, 2004 Journal of Japanese Society of Home Economics, Vol.45 No.7 p615 ～ 620, 1994 FISHERIES SCIENCE, 14 p921-928, 2002 Journal of Japanese Society for Taste and Smell, Vol.14 No.1 p9-20, 2007 JP-A-2005-245438 JP 2005-245438 A

It is an object of the present invention to provide a composition that maintains and enhances the umami of food, a production method, and a food material containing the same.

As a result of intensive studies, the present inventors have found that the problem can be solved by using a umami component, particularly a nucleic acid-based umami component and the tetrapeptide, and have completed the present invention. Rather than using an umami component alone, it was found that the umami was enhanced and the richness increased, and the present invention was completed. That is, the present invention includes 1, a tetrapeptide represented by SEQ ID NO: 1 Leu-Met-Lys-Pro (LMKP), a tetrapeptide represented by SEQ ID NO: 2 Ile-Lys-Gln-Asp (IKQD), SEQ ID NO: 3 Ala-Gln-Pro-Ala (
AQPA), a tetrapeptide represented by SEQ ID NO: 4 Ala-Gln-Pro-Thr (AQPT), or a derivative thereof, or a salt thereof. Composition, 2, a composition comprising a tetrapeptide of SEQ ID NOs: 1 to 4 and a nucleic acid-based taste ingredient, 3, a composition according to claim 1 or 2 used for imparting a rich taste function To do.

The composition used in the present invention contains 0.0001% by weight or more of a tetrapeptide containing at least one of LMKP, IKQD, AQPA or AQPT, and preferably contains 0.01% by weight or more.

In this specification, kokumi generally refers to something related to richness, strength, animal taste, umami, persistence, spread, depth, and the like. The nucleic acid-based umami component used in the present invention is 5
´-guanylic acid and 5´-inosinic acid.

A relatively large amount of the peptide of the present invention is contained in yeast. Examples of yeast include baker's yeast, brewer's yeast, and Torula yeast. Among them, Torula yeast contains a large amount.

The peptide used in the present invention can be prepared by a known method. For example, the peptide of the present invention can be obtained by a chemical synthesis method (for example, a solid phase method (for example, Fmac method). The amino acids constituting the peptide of the present invention may be L-form or D-form. But preferably
L body.

Furthermore, the peptide of the present invention can also be prepared by cutting out a peptide comprising the target amino acid sequence from the amino acid sequence of the protein containing the target amino acid sequence by a known means such as protease treatment. For example, examples of the protein containing the LMKP sequence include proteins derived from baker's yeast as shown in Table 1. In addition, any food containing the LMKP sequence, preferably a protein derived from Torula yeast, is preferably used as a substrate.
(table 1)

A person skilled in the art considers the sequence specificity of the protease, etc., to select a neutral protease (derived from Bacillus subtilis) suitable for excising a peptide comprising the target amino acid sequence from the amino acid sequence of the protein containing the target amino acid sequence. It can be selected appropriately. For example, in order to excise the LMKP sequence from the baker's yeast-derived sequence, use of a neutral protease (EC 3.4.24.4) can be mentioned. Commercially available proteases include protease N “Amano” G, which is an endo-type protease. Neutral protease (EC3.4.24.4) specifically hydrolyzes peptide bonds on the amino group side of hydrophobic amino acids when the enzyme concentration is low.

The reaction conditions used when the protein is hydrolyzed with a protease are not particularly limited, and can be appropriately selected by those skilled in the art according to common general technical knowledge. For example, in the case of a commercially available protease, the reaction conditions can be selected according to the instructions for use. Generally, 30-80 ° C
A reaction temperature of preferably 40 ° C. to 70 ° C. can be used. In general, reaction times of 2 to 30 hours, preferably 3 to 24 hours may be used. The reaction pH is preferably near the optimum pH of the protease to be used. The reaction stopping means is not particularly limited, and known means can be used. For example, heat treatment such as heating at 85 ° C. for 15 minutes or heating at 100 ° C. for 5 minutes can be mentioned.

After hydrolysis with protease, the target peptide can be obtained by purification by a known means. As known means, strong acid ion exchange treatment, octadecyl silica (ODC) resin, or the like can be used. As an example, a target peptide can be purified by adsorbing an aqueous peptide solution after protease treatment to an ODS resin and eluting it with an organic solvent of any concentration (for example, acetonitrile or the like). Alternatively, the target peptide can be purified by adsorbing the aqueous peptide solution after the protease treatment to a strongly acidic ion exchange resin and using an eluate (sodium chloride, potassium chloride) having a salt concentration of about 0.18 to 0.25M.

As described above, a peptide obtained by hydrolyzing a natural protein with a protease is more advantageous from the viewpoint of cost than a case where it is produced by a chemical synthesis method. Furthermore, it is considered that a peptide obtained by hydrolyzing a natural protein with a protease is safer for a living body. Therefore, the peptide thus obtained can be suitably used for foods and the like that require high safety for application to living bodies.

Derivatives of the peptides of the present invention can be obtained by the solid phase synthesis method (LACarpino, GYHan, J. Am.
em. Sci., 92, 5748 (1970)) and the like.

In the present invention, the above-mentioned specific peptides may be used alone or in any combination of two or more.

The blending amount of the specific peptides blended in the composition of the present invention is not particularly limited as long as the effect of the present application can be obtained, but it is usually preferable to be about 0.00001 to 50% by weight with respect to the food composition. Especially 0.00
It is preferable to blend 1 to 1% by weight.

The blending weight ratio between the peptides of the present invention and the nucleic acid-tasting component is not particularly limited as long as the effect of the present invention can be achieved. Usually, nucleic acid-based taste components (5′-guanylic acid and 5′-inosinic acid) 1 The specific peptide is 0.0001 to 1000 parts by weight, preferably 0.001 to 100 parts by weight, and more preferably 0.01 to 10 parts by weight with respect to parts by weight. With this blending ratio, it is easy to handle and easy to blend with other components.

Examples of foods that can be used or blended include fat and oil foods, bakery products, confectionery, noodles, processed rice products, processed wheat products, processed corn products, processed beans (soybeans, red beans, etc.), cereals (buckwheat noodles, sea breams, awa , Millet) processed products, dairy products, soups, beverages, seasonings, processed meat products, processed marine products, cooked and prepared foods, health foods, low calorie foods, foods for allergic patients, foods for infants, foods for the elderly, Examples thereof include beauty foods, medicinal foods, and further frozen foods, retort foods, instant foods, and canned foods.

Examples of the oil and fat food include margarine, shortening, whipped cream, cream, salad oil, custard cream, dip cream, fat spread, mayonnaise, tartar sauce, dressing, and oil frying.

Examples of the above bakery products include bread, white bread, sweet bread, side dish bread, French bread,
Examples include croissants, pies, castellas, sponge cakes, butter cakes, shoe confectionery, waffles, steamed bread, and fermented confectionery.

Examples of confectionery include snacks, donuts, biscuits, crackers, buns,
Japanese sweets, yokan, in the middle, uiro, dumplings, daifuku mochi, candy, gum, chocolate,
Examples include salmon, ice cream, soft cream, sherbet, popsicle, lact ice, ice confectionery, jelly, pudding, dessert, and topping.

Examples of the above processed rice products include cooked rice (including frozen cooked rice, sterile cooked rice, etc.), rice balls, rice ball rice cakes, rolled rice cakes, chili rice cakes, rice cakes, pilaf, tea pickles, doria, rice noodles, arabe, rice crackers, etc. Can be mentioned.

Examples of processed wheat products include cereal, udon, pizza, pasta, hoto, Chinese soba,
Yakisoba, champon, okonomiyaki, monja-yaki, piroshki, buns, cup noodles and their ingredients.

Examples of the corn processed product include corn flakes, corn snacks, popcorn and the like.

Examples of the processed soybean product include tofu, soy milk, soy milk beverage, yuba, deep-fried oil, deep-fried chicken, cancer pounding, red bean paste, miso, various bean dishes, and the like.

Examples of processed cereals (buckwheat, sea bream, awa, millet) and processed agricultural products (potato, sweet potato, taro, troroimo, etc.) include buckwheat, potato chips, sweet potatoes, potato fries, and various potato dishes. .

Examples of the dairy products include milk, processed milk, yogurt, whey drink, lactic acid bacteria drink, butter,
Examples include cheese and coffee whitener.

Examples of the soup include potage soup, consomme soup, stew, miso soup, soup, soup, curry and the like, and these may be added to these ingredients. Or you may add in the foodstuffs added directly to these soups like glutone.

Examples of the beverages include soft drinks, carbonated drinks, colas, juices, fruit juices, vegetable juices, tomato juices, shakes, sake, beer, sparkling wine, western liquor, wine, fruit liquors, cocktails, tea, tea, coffee , Cafe au lait, oolong tea, green juice, mineral water, water and the like.

Examples of the above-mentioned seasonings include soy sauce, fish sauce (squid crab soy sauce, sardine soy sauce, salted juice, nanpura, etc.), miso, jam, sauce, toaster sauce, tomato sauce, tomato ketchup, tomato paste, tomato puree, chili sauce, sauce , Pepper, pepper, garlic, ginger, vinegar, chili oil, tabasco, salt, various spices.

Examples of the processed meat product include hamburger, sausage, salami sausage, ham, meatball, meat dumpling, meat bun, dumpling, shumai, various meat dishes, and the like.

Examples of the processed fishery product include kamaboko, deep-fried sweet potato, tsumire, and kneaded products.

Examples of the health food or medicinal food include supplements, tablets, drinks, sports drinks and the like.

In the case of blending into food, the blending amount may be determined according to the purpose. For example, it is preferably 0.0001 to 50% by weight, particularly preferably 0.001 to 1% by weight based on the total amount of food.

In the composition of the present invention, the peptide or derivative thereof or a salt thereof is appropriately blended, for example, with carriers, base materials, additives, and the like that are usually used in the food field and the like within the scope of achieving the object of the present invention. Can be prepared. Examples of the carrier include sugars (glucose, maltose, mannitol, lactose, nonfat dry milk, dextrin, etc.), celluloses (eg, hydroxypropylcellulose, methylcellulose, crystalline cellulose), poorly water-soluble gums (eg, gum arabic, gum tragacanth) Etc.), cross-linked vinyl polymers, lipids and the like may be used alone or in combination of two or more. Particularly preferred for improving the flavor are dextrins with high DE and skim milk powder.

Examples of feeds that can be used or blended include livestock feed, pet feed, seafood feed,
For example, feed for farmed fish.

In the case of blending into feed, the blending amount may be determined according to the purpose. For example, it is preferably blended in an amount of 0.01 to 10% by weight, particularly 0.01 to 0.5% by weight based on the total amount of the feed.

When the tetrapeptide of the present invention is used in foods, pharmaceuticals, feeds, etc., a substance that gives some physiological activity to the body when ingested may be further blended. For example, amino acids, peptides, lipids, polysaccharides, oligosaccharides, proteins, carbohydrates, dietary fiber, enzyme components and the like can be mentioned.

Specific examples of substances that give these physiological activities include polyunsaturated fatty acids (EPA / DHA),
Plant sterol that regulates serum cholesterol, and its esterified products, diacylglycerol, γ-linolenic, α-linolenic acid, linoleic acid, conjugated linoleic acid and other unsaturated fatty acids, beet fiber, corn fiber, psyllium seed coat, tea polyphenol, lecithin, Bonito peptide, sardine peptide, casein decapeptide, soybean isolate protein, lactic acid bacteria, gluconic acid, oligosaccharide. Others, chlorella, spirulina, propolis, chitin, chitosan, deoxyribonucleic acid, ribonucleic acid, ganoderma, agaricus, ginkgo biloba extract, citrus fruit, turmeric, garcinia, collagen, blueberry, aloe, saw palmetto, capsaitin, lutein, β-crypt Xanthine, renin, taurine, casein, collagen, glucosamine, casein phosphopeptide, theanine, gabba, aspartame, xylitol, recardent, alginic acid, sodium alginate, fucodan, chondroitin, hyaluronic acid, cellulose, peptin, indigestible dextrin, glucomannan, inulin , Fructan, cyclofructan, difructose, levan, mucin, flavonoid, polyphenol, catechin, tannin, anthocyanin, rutin, Lucetin, Soy isoflavone, Soy saponin, Soy globulin, Chlorogenic acid, Citric acid, Lactic acid, Malic acid, Capsaicin, Somaliignan allicin, Caffeine, Chlorophyll, Nattokinase, β-lactoglobulin, Plant fermentation enzyme, Mevalonic acid, Chlorophyll, Royal jelly, Ginseng, prunes,
Chamomile, thyme, sage, peppermint, lemon palm, mallow, oregano, vitamins, calcium fortifiers such as calcium-containing compounds, iron fortifiers such as iron-containing compounds, mineral fortifiers containing essential minerals, and animal and plant extract ingredients And physiologically active components of naturally derived components such as seaweed extract components and herbal medicine components.

As vitamins that can be added, those listed as fortifiers in food additives can be used, such as vitamin B12, folic acid, pantothenic acid, niacin, biotin, vitamin C, vitamin D, vitamin E, vitamin K, these derivatives, oil-coated vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, pantothenic acid, niacin,
Biotin, vitamin C, coenzyme Q10, etc. Specific examples of vitamin derivatives include thiamine hydrochloride, thiamine sulfate, dibenzoyl thiamine, dibenzoyl thiamine nitrate, thiamine sulfate, dibenzoyl thiamine as vitamin B1, and riboflavin butyrate and riboflavin 5′-phosphate as vitamin B2. Examples include sodium ester, pyridoxine hydrochloride as vitamin B6, and L-ascorbic acid stearate and sodium L-ascorbate as vitamin C.

EXAMPLES The present invention will be described in detail below based on examples, but the present invention is not limited to these examples.

Example 1 (1) Enzyme-treated Candida utilis KLS-0582 (FERM from yeast cells)
After adjusting 1000 ml of 10% cell suspension of P-7396) to pH 3.5 with sulfuric acid and treating at 60 ° C. for 30 minutes, the cells were collected by centrifugation and washed with water. After adjusting the cell concentration to 10% with water and suspending it, 90%
The mixture was heated for 30 minutes to completely inactivate the intracellular enzyme, and 0.5 g of cell wall lytic enzyme (Tunicase Yamato Kasei) was added at 40 ° C. and pH 7.0 and reacted for 4 hours. The cell residue is removed by centrifugation, and the resulting supernatant is adjusted to pH 5, 0.1 g of ribonuclease Amano D (Amano Enzyme) is added, reacted at 65 ° C. for 4 hours, and then deaminase (Amano Enzyme) 0.3 g was added and reacted at 50 ° C. for 4 hours. After completion of the reaction, the reaction solution was heated to deactivate the added enzyme, and then concentrated, spray dried, and 30 g of yeast extract were obtained. This yeast extract is 5´-GMP5.5 wt%, 5´-IMP5.5 wt%
Contained. The peptide content in the yeast extract is 92.7% by weight (total amino acid content: 1
7.8 wt%, free amino acid content: 1.3 wt%).

Fractionation of the peptide of the present invention and sensory test of each fraction (1) Peptide fractionation method 100 ml of the 5% by weight yeast extract aqueous solution described above was added to ODS resin (Chromatorex, 100-200mesh, manufactured by Fuji Silysia Chemical)
The column was added to a column (124 ml) and eluted stepwise in the order of 100 ml of water → 200 ml of 5% EtOH → 200 ml of 10% EtOH → 200 ml of 100% EtOH, and fractionated every 200 ml. Flow conditions are column 25 ° C, flow rate 5ml / min
. Fraction eluted with 10% EtOH peptide with almost complete removal of nucleic acid content (10% EtOH fraction)
200 ml was concentrated and dried by a rotary evaporator and concentrated to 50 ml. The concentrated solution was added to a gel filtration resin (Sepahadex G-25, manufactured by Amersham Bioscience) column (195 ml), eluted with distilled water, and fractionated to 440 ml every 10 ml. As flow conditions, column temperature 25 ° C, flow rate 10 ml
/ min. The absorbance at a wavelength of 280 nm was measured for the fraction, and the chromatogram shown in FIG. 1 was obtained. Based on the chromatogram, 0-180 ml of eluate is G1 fraction, 180-220 ml is G2 fraction, 220-320 ml is G3 fraction
The fraction was used. The molecular weight was measured under the conditions shown in Table 2. FIG. 2 shows a chromatogram of the sample and 10% EtOH fraction measured under the conditions shown in Table 2. From the retention time of the specimen, it was estimated that the G1 fraction was tens of thousands to 700, the G2 fraction was 700 to 400, and the G3 fraction was less than 400.

Table 2 shows the conditions for molecular weight measurement.
(Table 2)

Table 3 shows the ratio (weight / weight%) of each fraction in the total amount of yeast extract.
(Table 3)

Table 4 shows the proportion (weight / weight%) of the G1 to G3 fractions in the entire 10% EtOH fraction.
(Table 4)

Sensory evaluation test for each fraction Synergy effect test with nucleotides 5'-Guanylate Na (5'-GMP, manufactured by Yamasa Shoyu Co., Ltd.) 200ppm solution, so that the above fraction is 400ppm in terms of solid content Added to. Sensory evaluation was performed by five panelists. The case where only the 5′-GMP solution was drunk was used as a control, and the strength of the aftertaste was evaluated as an index. The meanings of the symbols in Table 5 are as follows: ×: Less than control, Δ: Same level as control, ○: Stronger aftertaste than control, A: Stronger aftertaste than control. In the G2 fraction addition group, after tasting the taste of 5′-guanylic acid, the taste remained for a while and spread.

(Table 5)

(図２）

(1) Purification and identification of peptide of G2 fraction Four types of peaks detected at 280 nm were collected from the G2 fraction by reversed-phase high performance liquid chromatography. (2) Identification of peptides in G2 fraction The peaks in G2 fraction were subjected to peptide sequence by Edman degradation method, and the respective sequences were determined. Leu-Met-Lys-Pro , Ile-Lys-Gln-Asp, Ala-Gln-Pro-Thr, and Ala-Gln-Pro-Ala. The G2 fraction contained 0.057 wt%, 0.024 wt%, 0.001 wt% and 0.002 wt%, respectively.
(Figure 1)

(Figure 2)

The present invention provides a useful new composition that enhances umami and imparts a rich feeling. The composition of the present invention can be used to enhance the umami component, and can maintain and enhance umami by adding a small amount to a food or drink. Although the novel tetrapeptide composition of the present invention itself is tasteless, it can impart richness, a feeling of fat, and a rich feeling only by adding it to food.

Claims (3)

Tetrapeptide represented by SEQ ID NO: 1, a tetrapeptide represented by SEQ ID NO: 2, compositions containing the tetrapeptide represented by tetrapeptides and SEQ ID NO: 4, represented by SEQ ID NO: 3. The composition of Claim 1 containing the tetrapeptide of sequence number 1-4 and the nucleic acid type | system | group taste component are contained. The composition of Claim 1 or 2 used in order to give kokumi.