WO2011081186A1 - Flavor-enriching agent - Google Patents

Flavor-enriching agent Download PDF

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Publication number
WO2011081186A1
WO2011081186A1 PCT/JP2010/073722 JP2010073722W WO2011081186A1 WO 2011081186 A1 WO2011081186 A1 WO 2011081186A1 JP 2010073722 W JP2010073722 W JP 2010073722W WO 2011081186 A1 WO2011081186 A1 WO 2011081186A1
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WO
WIPO (PCT)
Prior art keywords
glu
gly
food
nva
taste
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Application number
PCT/JP2010/073722
Other languages
French (fr)
Japanese (ja)
Inventor
貴志 宮木
直宏 宮村
恵 金子
裕右 網野
礼子 安田
譲 江藤
高穂 田島
Original Assignee
味の素株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Application filed by 味の素株式会社 filed Critical 味の素株式会社
Priority to AU2010339306A priority Critical patent/AU2010339306B2/en
Priority to JP2011547717A priority patent/JP5850399B2/en
Priority to KR1020127019882A priority patent/KR101512627B1/en
Priority to CN201080063730.6A priority patent/CN102753041B/en
Priority to MX2012007244A priority patent/MX2012007244A/en
Priority to NZ601030A priority patent/NZ601030A/en
Priority to CA2783415A priority patent/CA2783415C/en
Priority to RU2012132448/10A priority patent/RU2532106C2/en
Priority to SG2012048146A priority patent/SG181978A1/en
Publication of WO2011081186A1 publication Critical patent/WO2011081186A1/en
Priority to IL220527A priority patent/IL220527A0/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
    • A23L27/22—Synthetic spices, flavouring agents or condiments containing amino acids containing glutamic acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/175—Amino acids

Definitions

  • the present invention relates to a body taste imparting agent and a composite body taste imparting agent comprising a peptide exhibiting CaSR agonist activity.
  • the present invention also relates to a food composition containing a peptide having CaSR agonist activity at a certain concentration or more.
  • Patent Document 1 describes that ⁇ -Glu-X-Gly (X is an amino acid or an amino acid derivative) is a compound having CaSR agonist activity, etc., but ⁇ -Glu-Nva-Gly is described. Is not described in detail in the examples, and is not disclosed specifically. In addition, the content of patent document 1 and 2 shall be contained in description of this specification.
  • the present invention searches for many variation compounds having CaSR agonist activity and has a more excellent body taste imparting action, in particular, an aftertaste type and high potency body taste imparting action, and has excellent stability. It is an object of the present invention to find a substance capable of imparting a taste, and to provide a kokumi imparting agent comprising the substance and a complex kokumi imparting agent comprising the substance in combination with another substance having CaSR agonist activity. It is another object of the present invention to provide a food composition containing the substance at a certain concentration.
  • ⁇ -Glu-Nva-Gly L- ⁇ -glutamyl-L-norvalyl-glycine
  • CaSR agonist activity a substance that is a carboxylic acid
  • ⁇ -Glu-Nva-Gly has an imparting effect, and in particular, its taste pattern can impart a rich taste that is a medium aftertaste type.
  • ⁇ -Glu-Nva-Gly has an extremely high titer of 10 times or more compared with the same tripeptide ⁇ -Glu-Val-Gly, and has excellent stability. And it discovered that the preferable taste pattern that a middle aftertaste was strong was shown.
  • ⁇ -Glu-Nva-Gly can be a useful body taste imparting agent by itself. Further, it has been found that by adding ⁇ -Glu-Nva-Gly, a preferable food composition with enhanced richness can be obtained. Furthermore, it discovered that it could become a complex body taste imparting agent formed by using this substance together with another substance having CaSR agonist activity, and completed the present invention.
  • the present invention provides a rich taste imparting agent comprising ⁇ -Glu-Nva-Gly.
  • the present invention also provides a food composition containing ⁇ -Glu-Nva-Gly (hereinafter also referred to as “the food composition of the present invention”).
  • the present invention also relates to (a) ⁇ -Glu-Nva-Gly, (b) ⁇ -Glu-X-Gly (X represents an amino acid or amino acid derivative, except Nva), ⁇ -Glu-Val- Y (Y represents an amino acid or amino acid derivative), ⁇ -Glu-Nva, ⁇ -Glu-Abu, ⁇ -Glu-Ala, ⁇ -Glu-Gly, ⁇ -Glu-Cys, ⁇ -Glu-Met, ⁇ - Glu-Thr, ⁇ -Glu-Val, ⁇ -Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, ⁇ -Glu-Met ( O), ⁇ -Glu- ⁇ -Glu-Val, ⁇ -Glu-Val-NH 2 , ⁇ -Glu-Val-ol, ⁇ -Glu-Ser, ⁇ -Glu-Tau, ⁇ -Glu-Cys (S- Me) (
  • the present invention has a very excellent body taste imparting action, in particular, a unique medium aftertaste type having a profile whose taste pattern is shown in FIG.
  • a rich body taste imparting agent and a complex body taste imparting agent that can be produced easily and at low cost can be provided.
  • the outstanding food composition which contains the substance which has the outstanding richness imparting effect
  • the rich taste imparting agent of the present invention can impart a fat-like richness and smoothness to the taste of low-fat foods, so even if the fat content in fat-containing foods is reduced, it is the same as the original food Can maintain a rich sense of health and can be made into a health-conscious food. Examples of such foods include meat-containing foods and dairy products.
  • a food containing the richness-imparting agent of the present invention is eaten, there is an advantage that a fat-like richness and smoothness can be felt afterwards without being eaten.
  • FIG. 1 shows a taste profile of a medium aftertaste type body taste imparting agent.
  • ⁇ -Glu-Nva-Gly targeted in the present invention includes L- ⁇ -glutamyl-L-norvalyl-glycine formed by peptide bonding of three amino acids and / or a salt thereof, particularly an edible salt. Since ⁇ -Glu-Nva-Gly has an excellent body taste imparting effect, it can be used as a body taste imparting agent. ⁇ -Glu-Nva-Gly is 0.1 ppb to 99.9% by mass, preferably 1 ppb to 10% by mass, more preferably 0.01 ppm to 1% by mass with respect to the weight of the food composition imparting a rich taste. % Can be added and used.
  • another aspect of the present invention relates to a food composition containing ⁇ -Glu-Nva-Gly, preferably a food composition containing 0.1 ppb to 99.9% by mass of ⁇ -Glu-Nva-Gly. . More preferably, the present invention relates to a food composition containing 0.01 to 50 ppm by weight of ⁇ -Glu-Nva-Gly.
  • the body taste imparting agent of the present invention is composed of amino acids such as sodium glutamate (MSG), nucleic acids such as inosine monophosphate (IMP), inorganic salts such as sodium chloride,
  • amino acids such as sodium glutamate (MSG)
  • nucleic acids such as inosine monophosphate (IMP)
  • inorganic salts such as sodium chloride
  • the term “kokumi” means five basic tastes represented by sweet taste, salty taste, sour taste, bitter taste, and umami. It means a taste that cannot be expressed in terms of basic taste, as well as thickness, thickness (mounthfulness), continuity, harmony, etc. The taste is also enhanced.
  • “kokumi impartation” means to enhance the five basic tastes expressed by sweetness, salty taste, acidity, bitterness, umami, and to give a taste around the basic tastes such as thickness, spread, and unity. Say. This can also be expressed as a taste enhancing action. Therefore, ⁇ -Glu-Nva-Gly, which is the body taste imparting agent of the present invention, can also be expressed as a flavor enhancer.
  • ⁇ -Glu-Nva-Gly which is a body taste imparting agent of the present invention can be used as a sweetness enhancer, salty taste enhancer, sour taste enhancer, bitterness enhancer or umami enhancer.
  • the taste changes with the passage of time after eating, but they are called an initial taste, a middle taste, and an after taste in order from immediately after eating.
  • taste, medium and aftertaste are tastes to be felt from 0 to 2 seconds, from 2 to 5 seconds, and after 5 seconds, respectively, after eating. Further, the period from 0 to 5 seconds is called “first taste”, and the period from about 2 seconds to about 30 seconds is called “medium after taste” (see FIG. 1).
  • CaSR means a calcium sensing receptor (Calcium® Sensing® Receptor), which belongs to the class C of the 7-transmembrane receptor, and is also referred to as a calcium receptor.
  • the “CaSR agonist” means a substance that binds to the CaSR and activates the CaSR.
  • activate CaSR means that a ligand binds to CaSR and activates a guanine nucleotide-binding protein to transmit a signal. The property of binding to CaSR and activating CaSR is referred to as “CaSR agonist activity”.
  • a test substance is added to a CaSR activity measurement system for measuring CaSR activity, and the CaSR activity is measured.
  • the CaSR activity when the test substance is added is compared with the CaSR activity when the test substance is not added.
  • the measurement of CaSR activity can be performed using, for example, a measurement system using cells that express CaSR.
  • the cell may be a cell that endogenously expresses CaSR or a recombinant cell into which a CaSR gene has been introduced exogenously.
  • the CaSR activity measurement system can detect binding (reaction) between an activator and CaSR when an extracellular ligand (activator) specific to CaSR is added to the cell expressing CaSR. It can be used without particular limitation as long as it can transmit a detectable signal in the cell in response to the binding (reaction) between the activator and CaSR.
  • CaSR activity is detected by reaction with the test substance, it is determined that the test substance has CaSR stimulating activity.
  • CaSR examples include human CaSR encoded by the human CaSR gene registered under GenBank Accession No. NM_000388.
  • CaSR is not limited to the protein encoded by the gene of the above sequence, and as long as it encodes a protein having a CaSR function, it is 60% or more, preferably 80% or more, more preferably 90% or more. It may be a protein encoded by a homologous gene.
  • the CaSR function can be examined by expressing these genes in cells and measuring changes in current and intracellular calcium ion concentration when calcium is added.
  • the origin of the CaSR is not particularly limited, and examples include CaSR derived from any animal including mice, rats, dogs and the like as well as the human CaSR.
  • the CaSR activity can be confirmed using a living cell expressing CaSR or a fragment thereof, a cell membrane expressing CaSR or a fragment thereof, an in vitro system containing a protein of CaSR or a fragment thereof, or the like.
  • An example using living cells is shown below, but is not limited thereto.
  • CaSR is expressed in cultured cells such as Xenopus oocytes, hamster ovary cells, and human fetal kidney cells. This can be achieved by introducing a plasmid carrying a foreign gene into which a CaSR gene has been cleaned and introducing the plasmid state or cRNA using it as a template.
  • an electrophysiological technique or a fluorescent indicator reagent for increasing intracellular calcium can be used.
  • CaSR expression is first confirmed by a response with calcium or a specific activator.
  • An oocyte in which an intracellular current is observed or a cultured cell in which fluorescence of a fluorescent indicator reagent is observed is used with respect to calcium having a concentration of about 5 mM. The concentration dependence is measured by changing the calcium concentration.
  • the test substance is prepared to about 1 ⁇ M to 1 mM, added to the oocyte or cultured cell, and the CaSR activity of the test substance is measured by measuring the CaSR activity in the presence of the test substance. taking measurement.
  • examples of the CaSR agonist activity test include, but are not limited to, the tests shown in the test examples of the present specification.
  • the amino acid or peptide used in combination with ⁇ -Glu-Nva-Gly in the complex rich taste imparting agent of the present invention is ⁇ -Glu-X-Gly (X represents an amino acid or amino acid derivative other than Nva), ⁇ -Glu- Val-Y (Y represents an amino acid or amino acid derivative), ⁇ -Glu-Nva, ⁇ -Glu-Abu, ⁇ -Glu-Ala, ⁇ -Glu-Gly, ⁇ -Glu-Cys, ⁇ -Glu-Met, ⁇ -Glu-Thr, ⁇ -Glu-Val, ⁇ -Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, ⁇ -Glu- Met (O), ⁇ -Glu- ⁇ -Glu-Val, ⁇ -Glu-Val-NH 2 , ⁇ -Glu-Val-ol, ⁇ -Glu-Ser, ⁇ -Glu-Tau, ⁇ -G 1 selected
  • Neutral amino acids acidic amino acids such as Asp and Glu, basic amino acids such as Lys, Arg and His, aromatic amino acids such as Phe, Tyr and Trp, homoserine, citrulline, ornithine, ⁇ -aminobutyric acid, norvaline, norleucine , Taurine and the like are also included. Further, it may be a non-natural (non-protein constituent) amino acid such as tert-leucine, cycloloinsine, ⁇ -aminoisobutyric acid, L-penicillamine and the like.
  • X may be any of the above amino acids or derivatives thereof, but an amino acid other than Cys or a derivative thereof is preferred.
  • ⁇ -Glu-Val-Gly, ⁇ -Glu-Abu-Gly, ⁇ -Glu-tLeu-Gly, ⁇ -Glu-Nva, ⁇ -Glu-Abu and the like are preferable.
  • the body taste imparting agent of the present invention is composed of ⁇ -Glu-Nva-Gly and has a unique medium aftertaste type excellent body taste imparting action having a profile as shown in FIG. It is preferably used in combination with a peptide having a different profile from ⁇ -Glu-Abu-Gly, ⁇ -Glu-Abu, etc.
  • an amino acid residue means the following amino acids.
  • amino acid derivatives are various derivatives of the above amino acids.
  • amino acids side chains such as special amino acids, unnatural amino acids, amino alcohols, terminal carbonyl groups, amino groups, cysteine thiol groups, and the like are substituted with various substituents. Substituted ones are mentioned.
  • substituents include an alkyl group, an acyl group, a hydroxyl group, an amino group, an alkylamino group, a nitro group, a sulfonyl group, and various protective groups.
  • Val-NH 2 Valinamide
  • Val-ol Valinol (2-amino- 3-methyl-1-butanol) and the like.
  • ⁇ -Glu-Cys (SNO) -Gly has the following structural formula
  • ⁇ -Glu-Met (O) and ⁇ -Glu-Cys (S-Me) (O (O) in the formula means a sulfoxide structure.
  • ( ⁇ ) of ⁇ -Glu means that another amino acid is bonded via a carboxyl group at the ⁇ position of glutamic acid.
  • the ⁇ -Glu-Nva-Gly and the amino acid or peptide used in combination with the ⁇ -Glu-Nva-Gly of the present invention may be a commercially available product, in addition, (1) a chemical synthesis method, or ( 2) Although it can be obtained by appropriately using a known method such as a method of synthesizing by an enzymatic reaction, chemical synthesis is simpler. Since ⁇ -Glu-Nva-Gly used in the present invention has a short amino acid residue of 3 residues, the chemical synthesis method is relatively simple and industrially advantageous.
  • the oligopeptide is synthesized or semi-synthesized using a peptide synthesizer.
  • the chemical synthesis method include a peptide solid phase synthesis method.
  • the peptide thus synthesized can be purified by conventional means such as ion exchange chromatography, reverse phase high performance liquid chromatography, affinity chromatography and the like. Such peptide solid phase synthesis methods, and subsequent peptide purification, are well known in the art.
  • ⁇ -Glu-Nva-Gly and an amino acid or peptide used in combination therewith by an enzymatic reaction for example, the method described in International Publication Pamphlet WO 2004/011653 is used. May be. That is, an amino acid or dipeptide in which the carboxyl terminus of one amino acid or dipeptide is esterified or amidated, and an amino acid in which the amino acid is free (for example, an amino acid in which the carboxyl group is protected) are combined in the presence of a peptide-forming enzyme. It is also possible to produce by producing the dipeptide or tripeptide produced by reacting in the above.
  • the peptide-forming enzyme examples include a culture of a microorganism having the ability to produce a peptide, a microbial cell separated from the culture, a treated product of the microorganism, or a peptide-generating enzyme derived from the microorganism.
  • the matters described in WO 2004/011653 are included in the description of this specification.
  • the peptides used in the present invention may be present in plants such as vegetables and fruits, microorganisms such as yeast, and other natural products. If they exist in nature, they can be extracted from these and used.
  • the kokumi imparting agent or the complex kokumi imparting agent of the present invention can be used as a seasoning as it is or by mixing it with a carrier and other seasoning ingredients that are acceptable for food and drink.
  • seasoning materials include, for example, flavorings, sugars, sweeteners, dietary fibers, vitamins, amino acids such as sodium glutamate (MSG), nucleic acids such as inosine monophosphate (IMP), and inorganic substances such as sodium chloride. Examples thereof include organic acids such as salts and citric acid, and various yeast extracts.
  • the low fat food preferable as a food composition containing the rich taste imparting agent or the complex rich taste imparting agent of the present invention is a food originally containing fat, and particularly a food having a reduced fat content.
  • fat is synonymous with “oil and fat”, includes both solid and liquid, and may be either animal fat or vegetable fat.
  • Such low-fat foods include dairy products such as milk, yogurt, butter and cream, margarine, milk for coffee, foods containing animal oils and / or vegetable oils such as sauces, roux, emulsified foods such as dressings and mayonnaise, etc. And various curries and stews containing cooked meat, and various soups containing meat extracts.
  • steaks and grilled meat made of cooked low-fat beef and baked snacks that are not subjected to normal frying are also included.
  • the low-fat food those whose normal fat content is 1/2 to 1/3 are preferable.
  • the rich taste-imparting agent of the present invention in the low-fat foods, when these foods are eaten, the fat-like richness and smoothness can be felt next, not first.
  • milk and yogurt normal products are 3-4% fat, while zero fat products (about 0.1% fat) are also known.
  • the savoriness imparting agent or the complex savoriness imparting agent of the present invention is also effective for these zero fat products.
  • the present invention also provides a food composition containing ⁇ -Glu-Nva-Gly and pork ingredients.
  • the food containing the pork raw material is not particularly limited, and examples thereof include pork extract, sausage, and instant noodle soup.
  • the content of the pork raw material is not particularly limited, and examples thereof include a food composition of the present invention that is about 0.005 to 80% by weight.
  • the present invention also provides a food composition containing ⁇ -Glu-Nva-Gly and a beef raw material.
  • a foodstuff containing a beef raw material For example, beef extract, corn beef, beef use soup, beef use sauce, etc. are mentioned.
  • a food composition may be about 0.005 to 80% by weight.
  • ⁇ -Glu-Nva-Gly and the amino acid or peptide used together in the present invention also include a salt form.
  • the salt may be a pharmacologically acceptable edible salt.
  • acidic groups such as groups, ammonium salts, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnesium, aluminum salts, zinc salts, triethylamine, ethanolamine, morpholine
  • alkali metals such as sodium and potassium
  • alkaline earth metals such as calcium and magnesium
  • aluminum salts such as calcium and magnesium
  • zinc salts triethylamine, ethanolamine, morpholine
  • organic amines such as pyrrolidine, piperidine, piperazine and dicyclohexylamine
  • salts with basic amine acids such as arginine and lysine.
  • salts with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, hydrobromic acid, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, Salts with organic carboxylic acids such as tannic acid, butyric acid, hibenzic acid, pamoic acid, enanthic acid, decanoic acid, teocric acid, salicylic acid, lactic acid, oxalic acid, mandelic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p And salts with organic sulfonic acids such as toluenesulfonic acid.
  • inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, hydrobromic acid, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, succin
  • the rich taste imparting agent, food composition, or complex rich taste imparting agent of the present invention can be used in any form without limitation on physical properties such as dry powder, paste, and solution.
  • the rich taste imparting agent, food composition, or complex rich taste imparting agent of the present invention can be used by blending it with various foods and beverages such as foods, beverages and seasonings.
  • the amount of the amino acid or peptide used in combination is not particularly limited as long as the desired effect is obtained, but the amount of ⁇ -Glu-Nva-Gly and / or the amount of amino acid or peptide may be food, beverage or seasoning, etc. From about 0.1 ppb to 99.9% by weight, preferably from 1 ppb to 10% by weight, and more preferably from about 0.01 ppm to 1% by weight.
  • any solid or liquid carrier that is acceptable for foods and beverages may be further blended.
  • the carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, gelatin, albumin, amino acid, water, and physiological saline. Water etc. are mentioned.
  • the seasoning raw material may be any seasoning raw material used in the art and is not particularly limited, but more specifically, the above-mentioned ones are already mentioned.
  • the content of any of the above carriers and other seasoning ingredients is not particularly limited.
  • the yeast extract is not particularly limited in any of the cells from which it is derived, its culture conditions, and the extraction treatment method, and any yeast extract can be used. Further, heat treatment, enzyme treatment, concentration, powder It may be one that has been processed.
  • the present invention also provides a method for producing various foods and drinks, characterized in that ⁇ -Glu-Nva-Gly is added to the production intermediate product of various foods and drinks so that 1 mass ppb to 99.9% by mass is contained. provide.
  • various foods and drinks low-fat foods are preferable.
  • This invention also provides the manufacturing method of various food-drinks characterized by adding the food composition of this invention to the manufacture intermediate goods of various food-drinks.
  • various foods and drinks low-fat foods are preferable.
  • a taste enhancer comprising ⁇ -Glu-Nva-Gly is used as a raw material for foods and beverages (for example, umami raw material, protein hydrolyzate, animal meat extract).
  • a method for producing food or drink or a food intermediate for producing food or drink which includes a step of adding to and mixing, and, if necessary, a step of further cooking the resulting food or drink raw material mixture, is preferred.
  • the step of adding and mixing the taste enhancer composed of ⁇ -Glu-Nva-Gly to the raw material for the food and drink, the concentration of ⁇ -Glu-Nva-Gly in the intermediate product of the food and drink is 0.01-999900 ppm by weight
  • it includes a step of adjusting to 0.1 to 200,000 ppm by weight.
  • the intermediate product of the food and drink is added to another food and drink raw material (for example, agricultural products, marine products, livestock meat, dairy products, or processed foods thereof), and the resulting food and drink ⁇ -Glu-Nva-Gly It is preferable to further include a step of adjusting the concentration to 0.01 to 50 ppm by weight, preferably 0.05 to 20 ppm by weight.
  • the step of adding and mixing the taste enhancer comprising ⁇ -Glu-Nva-Gly to the food / beverage product raw material has a concentration of ⁇ -Glu-Nva-Gly of the food / beverage product of 0.01 to 50 ppm by weight, preferably 0 It is preferable to include a step of setting the content to 0.05 to 20 ppm by weight.
  • food / beverage products are the foodstuffs containing a pork raw material. In this case, it is preferable to contain 0.01 to 50 ppm by weight of ⁇ -Glu-Nva-Gly, 0.005 to 80% by weight of pork ingredients, and other food ingredients.
  • food / beverage products are the foodstuffs containing a beef raw material. In this case, it is preferable to contain 0.01 to 50 ppm by weight of ⁇ -Glu-Nva-Gly, 0.005 to 80% by weight of beef ingredients, and other food ingredients.
  • the target foods of the present invention are foods such as ice cream, honey, marmalade, and strawberry jam (sweet foods) such as desserts and confectionery (sweet foods), and salty tastes such as chicken soup
  • foods such as processed foods, side dishes and snacks (savory foods) are also preferred.
  • the reaction solution was kept at 0 ° C., and triethylamine (Et 3 N, 3.13 ml, 1.1 eq, 22.4 mmol), HOBt ⁇ H 2 O (1-Hydroxybenzotriazole hydrate, 3.44 g, 1.1 eq, 22.4 mmol) and WSC ⁇ HCl (1 -Ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride, 4.30 g, 1.1 eq, 22.4 mmol) was added. The temperature of the reaction solution was gradually raised and stirred at room temperature overnight (16 hours).
  • the reaction mixture was concentrated under reduced pressure, ethyl acetate (150 ml) was added to the residue, and the organic layer was washed twice with water (50 ml) and 5% aqueous citric acid solution (50 ml), saturated brine (50 ml), 5
  • the extract was washed twice with an aqueous sodium hydrogen carbonate solution (50 ml) and with saturated brine (50 ml), and dried over anhydrous magnesium sulfate. Magnesium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure.
  • Boc-Nva-Gly-OBzl (6.88 g, 18.9 mmol) as crystals.
  • Boc-Nva-Gly-OBzl (6.88 g, 18.9 mmol) was added 4N HCl / dioxane solution (94.5 ml), and the mixture was stirred at room temperature for 1 hour.
  • Dioxane was removed by concentration under reduced pressure, and the operation of adding n-hexane (30 ml) to the residue and concentrating under reduced pressure was repeated three times to obtain H-Nva-Gly-OBzlHCl in a quantitative yield.
  • H-Nva-Gly-OBzlHCl was dissolved in methylene chloride (130 ml), and the reaction solution was kept at 0 ° C.
  • Z-Glu-OBzl N- ⁇ -Carbobenzoxy-L-glutamic acid ⁇ -benzyl ester, 7.03 g, 18.9 mmol
  • triethylamine (2.90 ml, 1.1 eq, 20.8 mmol
  • HOBt ⁇ H 2 O (3.20 g, 1.1 eq, 20.8 mmol)
  • WSC.HCl (3.98 g, 1.1 eq, 20.8 mmol) were added.
  • the temperature of the reaction solution was gradually raised and stirred at room temperature overnight (16 hours).
  • the reaction mixture was concentrated under reduced pressure, ethyl acetate (1000 ml) was added to the residue, and the organic layer was washed twice with water (100 ml) and 5% aqueous citric acid solution (100 ml), saturated brine (100 ml), 5
  • the extract was washed twice with an aqueous sodium hydrogen carbonate solution (100 ml) and with saturated brine (100 ml), and dried over anhydrous magnesium sulfate. After the solution was heated to 50 ° C., magnesium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. When crystals began to appear, n-hexane was added to sufficiently precipitate the crystals.
  • CaSR expression plasmid was prepared as follows. Based on the DNA sequence (CaSR (calcium receptor): NM_000388, SEQ ID NO: 1, 2) registered in NCBI, synthetic oligo DNA (forward primer (SEQ ID NO: 3: ACTAATACGACTCACTATAGGGACCATGGCATTTTATAGCTGCTGCTGG)) and reverse primer (SEQ ID NO: SEQ ID NO: 1) 4: TTATGAATTCACTACGTTTTCTGTAACAG) was synthesized.
  • CaSR calcium receptor
  • PCR was carried out under the following conditions using cDNA derived from human kidney (manufactured by Clontech) as a material and the above primers and Pfu Ultra DNA Polymerase (manufactured by Stratagene). After 3 minutes at 94 ° C., 35 times of 94 ° C. for 30 seconds, 55 ° C. for 30 seconds and 72 ° C. for 2 minutes were repeated 35 times, followed by reaction at 72 ° C. for 7 minutes. After agarose electrophoresis and staining with a DNA staining reagent, it was detected whether or not amplification was performed by UV irradiation. In addition, the chain length of the PCR product was confirmed by comparison with a DNA marker of known size that was electrophoresed simultaneously.
  • Plasmid vector pBR322 was cleaved with restriction enzyme EcoRV (Takara), and the gene fragment amplified by PCR was ligated to the cleavage site using Ligation kit (Promega).
  • the Escherichia coli DH5 ⁇ strain was transformed with this reaction solution, and a transformant carrying a plasmid in which the PCR amplification product was cloned was selected, and the PCR amplification product was further confirmed by DNA nucleotide sequence analysis.
  • a human CaSR expression plasmid hCaSR / pcDNA3.1 was prepared.
  • Assay Buffer 146 mM NaCl, 5 mM KCl, 1 mM MgSO 4 , 1 mg / ml Glucose, 20 mM HEPES (pH 7.2) 200 ul / well of Ca 2+ fluorescent indicator Calcium 4 Assay Kit (Molecular Devices) dissolved in 0.75 to 1.25 mM CaCl 2 ) was added and allowed to stand at 37 ° C. for 1 hour and then at room temperature for 10 minutes to incorporate the indicator. .
  • a test compound dissolved in 0.1% BSA-containing assay buffer was added to the 96-well plate at 50 ⁇ l / well, and the change in fluorescence intensity was measured with FLEX Station (Molecular Devices) for 3 minutes.
  • Example 1 Evaluation of kokumi imparting activity
  • the strength of kokumi imparting activity of ⁇ -Glu-Nva-Gly was examined by a quantitative sensory evaluation test.
  • the quantitative sensory evaluation test was performed as follows. In a distilled water containing sodium glutamate (0.05 g / dl), inosinic acid monophosphate (0.05 g / dl), sodium chloride (0.5 g / dl), the compounds as a sample were 0.000001-0. The strength of the savoriness imparting activity when mixed at 1 g / dl was measured. About the sample which showed the acidity with respect to the additive-free control after sample dissolution, it was used according to the range of pH ⁇ 0.2 with respect to the additive-free control with NaOH.
  • control 0 points, strong: 3 points, very strong: 5 points, and ⁇ -Glu-Cys-Gly's pre-taste and after-taste were set to 3.0 points to make the scale clearer.
  • the “first taste” refers to a taste of 0 to 5 seconds after the mouth, and the aftertaste is a taste after that.
  • the test compounds exhibited a wide range of richness-imparting activity at the above addition concentrations.
  • Table 4 shows the results of typical concentrations. As a result, all the tripeptides other than ⁇ -Glu-Nva-Gly were about 10 times as high as glutathione ( ⁇ -Glu-Cys-Gly), but surprisingly, ⁇ -Glu -Nva-Gly was even higher and was shown to be 100 times more highly active.
  • ⁇ -Glu-Nva-Gly is about 100 times higher than ⁇ -Glu-Cys-Gly, and at least 10 times higher in taste-providing activity than ⁇ -Glu-Val-Gly. Furthermore, ⁇ -Glu-Nva-Gly is superior, with no off-flavors (such as aftertaste astringency), such as ⁇ -Glu-Ala-Gly, ⁇ -Glu-Abu-Gly, and ⁇ -Glu-tLeu-Gly. I understood that.
  • Example 2 Evaluation of kokumi imparting activity For ⁇ -Glu-Nva-Gly, the strength of kokumi imparting activity was examined by a quantitative sensory evaluation test in another evaluation item in order to clarify the middle aftertaste type.
  • the quantitative sensory evaluation test was performed as follows. In order to make the middle aftertaste easier to understand, the innocous acid monophosphate was not used in the evaluation solution, and the umami after the middle was reduced. That is, when the compounds as a sample were mixed at 0.000001 to 0.1 g / dl in distilled water containing sodium glutamate (0.1 g / dl) and sodium chloride (0.4 g / dl), The intensity of taste imparting activity was measured.
  • n 4.
  • the “prior taste” is a taste of 0 to 2 seconds after the mouth is contained, and the medium after taste is a taste after that.
  • the test compounds exhibited a wide range of richness-imparting activity at the above-mentioned added concentrations. Table 5 shows the results of typical concentrations.
  • ⁇ -Glu-Val-Gly was about 10 times as active as glutathione ( ⁇ -Glu-Cys-Gly), but ⁇ -Glu-Nva-Gly was even higher, slightly more than 100 times. It was shown to have a high activity.
  • ⁇ -Glu-Nva-Gly has an excellent kokumi imparting activity, has a taste of medium aftertaste, and is excellent with no off-flavors (such as astringent taste).
  • ⁇ -Glu-Nva-Gly is about 100 times higher than ⁇ -Glu-Cys-Gly, and has a kokumi imparting activity that is at least 10 times higher than ⁇ -Glu-Val-Gly. It can be used at low concentrations. Therefore, it is possible to provide a richness-imparting agent more easily and at a low cost, which is very advantageous from an industrial viewpoint.
  • Example 3 Evaluation of kokumi- taste-imparting activity in foods ⁇ -Glu-Nva-Gly is actually sensory-evaluated whether it is extremely effective compared to high-titer ⁇ -Glu-Val-Gly when used in foods. It was examined by testing. The sensory evaluation test was performed as follows. Commercially available ice cream, honey, marmalade, and strawberry jam were used as representative foods such as desserts and confectionery that mainly have sweetness (sweet foods) as foods that are thought to have a strong aftertaste.
  • ⁇ -Glu-Val-Gly As a representative of processed foods such as salty foods, side dishes, and snacks (savory foods), a commercially available chicken soup, 0.1% by weight of powdered powdered pepper and mashed potato paste, commercially available ginger, and 2% by weight of butter The paste added to the mashed potato was used. The amount of ⁇ -Glu-Val-Gly to be compared was set to 0.002% by weight where the effect was obvious. When ⁇ -Glu-Nva-Gly was mixed at 0.0000001 to 0.01% by weight, the enhancement of the overall taste (strength of kokumi imparting activity) was measured. The control is an additive-free food.
  • n 4. With respect to ⁇ -Glu-Val-Gly, the richness-enhancing activity was widely exhibited at the above-mentioned added concentrations.
  • Tables 7 and 8 show the results of concentrations that can be clearly compared. This result also shows that ⁇ -Glu-Nva-Gly has an additional 5-13 compared to ⁇ -Glu-Val-Gly, which is about 10 times as rich as glutathione ( ⁇ -Glu-Cys-Gly). It was shown to have a double strength and extremely high activity.
  • ⁇ -Glu-Nva-Gly was found to have promising kokumi imparting activity that enhances the overall taste in all foods characterized by a medium aftertaste from actual savory to sweet foods. Furthermore, it was shown that ⁇ -Glu-Nva-Gly has an extremely high body taste imparting activity of 5 to 13 times as much as ⁇ -Glu-Val-Gly in actual foods. Therefore, it is desired to improve the quality. However, even if the raw materials cannot be blended any more because of the quality stability, the quality can be improved in a very small amount by using ⁇ -Glu-Nva-Gly. Moreover, it is possible to provide a rich taste imparting agent at low cost.
  • Example 4 Effect of ⁇ -Glu-Nva-Gly on pork extract As for ⁇ -Glu-Nva-Gly, kokumi imparting activity activity is higher than ⁇ -Glu-Cys-Gly (glutathione) and ⁇ -Glu-Val-Gly After eating, it turned out that it strengthened from an early time. Therefore, ⁇ -Glu-Nva-Gly is significantly more effective than ⁇ -Glu-Cys-Gly (glutathione) and ⁇ -Glu-Val-Gly for pork extracts that are not completely medium-type and have a slightly stronger taste. It was examined by a sensory evaluation test. The sensory evaluation test was performed as follows.
  • a commercially available pork extract (solid content 55.1 wt%, salt content 9.3 wt%) was dissolved in hot water so as to be 5.0 wt% to prepare a pork extract solution.
  • ⁇ -Glu-Nva-Gly, ⁇ -Glu-Cys-Gly, or ⁇ -Glu-Val-Gly was mixed as a sample. The measurement was performed using a two-point discrimination test method.
  • Table 9 shows the number of panels in which ⁇ -Glu-Nva-Gly 0.0003% by weight was evaluated as “poke extract is stronger and more favorable without changing the balance between taste and flavor”. From these results, it is clear that ⁇ -Glu-Nva-Gly clearly shows that “Poke extract is strengthened and favored without changing the balance between taste and flavor” even at the equivalent strength value as in (1) and (3). It was shown that.
  • the pork extract does not change the balance between taste and flavor base. It has been found that it has a very specific effect of “strengthening and favoring”. Pork ingredients are widely used worldwide for seasonings, soups, processed meat products, cooked products, confectionery, snacks, etc. Therefore, ⁇ -Glu-Nva-Gly can improve foods at a lower cost and in a minute amount, and is very advantageous from an industrial viewpoint.
  • Example 5 Effect of ⁇ -Glu-Nva-Gly on Beef Extract With ⁇ -Glu-Nva-Gly, Kokumi imparting activity is eaten from ⁇ -Glu-Cys-Gly (glutathione) and ⁇ -Glu-Val-Gly Later, it turned out that it strengthened from an early time. Therefore, ⁇ -Glu-Nva-Gly is significantly more ⁇ -Glu-Cys-Gly (glutathione) and ⁇ -Glu-Val-Gly than beef extract, which is not a perfect middle-type but has a slightly faster taste and lasts until aftertaste. Effectiveness was examined by a sensory evaluation test. The sensory evaluation test was performed as follows.
  • a commercially available beef extract (solid content: 61.2 wt%, salt content: 12.2 wt%) was dissolved in hot water to 3.0 wt% to prepare a beef extract solution.
  • ⁇ -Glu-Nva-Gly, ⁇ -Glu-Cys-Gly, or ⁇ -Glu-Val-Gly was mixed as a sample. The measurement was performed using a two-point discrimination test method.
  • Table 10 shows the number of panels in which ⁇ -Glu-Nva-Gly 0.0003% by weight was evaluated as “a beef extract is stronger and preferable without changing the balance between taste and flavor”. From these results, it is clear that ⁇ -Glu-Nva-Gly clearly “strengthens and favors beef extract without changing the balance of taste and flavor” even with the same rich taste strength as in (1) and (3) It has been shown.
  • “beef extract” can be used without changing the balance between taste and flavor base. It has been found that it has a very specific effect of “enhancing and favoring”. Beef ingredients are widely used worldwide for seasonings, soups, processed meat products, cooked products, confectionery, snacks, etc. Therefore, ⁇ -Glu-Nva-Gly can improve foods at a lower cost and in a minute amount, and is very advantageous from an industrial viewpoint.

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Abstract

Disclosed is a flavor-enriching agent comprising a substance discovered by searching for a number of variation compounds having a CaSR-agonizing activity. Said substance exhibits a potent flavor-enriching action, particularly with respect to middle tastes, and exhibits an improved stability. Also disclosed is a composite flavor-enriching agent comprising said substance and another substance having a CaSR-agonizing activity. Specifically disclosed are a flavor-enriching agent comprising γ-Glu-Nva-Gly (L-γ-glutamyl-L-norvaline-glycine), and a composite flavor-enriching agent comprising said substance together with another substance having a CaSR-agonizing activity.

Description

コク味付与剤Kokumi imparting agent
　本発明は、ＣａＳＲアゴニスト活性を示すペプチドからなるコク味付与剤及び複合コク味付与剤に関する。又、本発明はＣａＳＲアゴニスト活性を示すペプチドを一定濃度以上含有する食品組成物に関する。 The present invention relates to a body taste imparting agent and a composite body taste imparting agent comprising a peptide exhibiting CaSR agonist activity. The present invention also relates to a food composition containing a peptide having CaSR agonist activity at a certain concentration or more.
　近年、食生活の多様化等により味覚に対する消費者の要求が高まってきており、これに伴い、甘味、塩味、酸味、苦味、うま味で表される５基本味だけでは表すことのできない、厚み・ひろがり・持続性・まとまりなど上記基本味の周辺の味をも増強した味覚である「コク味」を付与することのできる優れたコク味付与剤へのニーズが高まっている。
　一方、カルシウムセンシング受容体（Calcium Sensing Receptor：ＣａＳＲ）は、カルシウム受容体とも呼ばれるが、当該受容体シグナルは種々の生体内機能を調節し、ＣａＳＲアゴニスト活性を有する物質はコク味付与剤として用いることができる（特許文献１および２、非特許文献３）。 In recent years, consumer demand for taste has increased due to diversification of eating habits, etc., and with this, thickness, thickness, which cannot be expressed only with 5 basic tastes expressed by sweetness, salty taste, acidity, bitterness, umami taste, There is an increasing need for an excellent body taste imparting agent capable of imparting a “kokumi” taste that enhances the taste around the basic tastes such as spread, sustainability, and unity.
On the other hand, calcium sensing receptor (Calcium Sensing Receptor: CaSR) is also called calcium receptor, but the receptor signal regulates various in vivo functions, and a substance having CaSR agonist activity should be used as a body taste imparting agent. (Patent Documents 1 and 2, Non-Patent Document 3).
　上記の「コク味」には種々の呈味パターンが存在するが、中後味型であるコク味を付与することのできる、より高力価のコク味付与剤が求められている。またコク味を付与する物質は通常、食品等に用いられるため、安定性に優れることが求められる。 There are various taste patterns in the above-mentioned “kokumi”, but there is a demand for a higher-potency kokumi imparting agent capable of imparting a medium aftertaste type kokumi. Moreover, since the substance imparting richness is usually used in foods and the like, it is required to have excellent stability.
　従って、ＣａＳＲアゴニスト活性を有する多くのバリエーション化合物を探索し、より優れたコク味付与作用、特に先味型のコク味付与作用を有し、かつ安定性に優れ簡便かつ低コストで生産することが可能なコク味を付与するこのできる物質を見出し、該物質からなるコク味付与剤、及び該物質を他のＣａＳＲアゴニスト活性を有する物質と併用してなる複合コク味付与剤を提供することが求められている。
　一方、γ－グルタミンをＮ末端に有するいくつかのγ－グルタミルペプチドについては、酵素活性の研究等において基質として合成された例は知られているが（特許文献３、非特許文献１、２）、γ-Ｇｌｕ－Ｎｖａ－Ｇｌｙが食品用途に実際に用いられた例や天然に存在した例は知られていない。
　また、前記特許文献１には、γ－Ｇｌｕ－Ｘ－Ｇｌｙ（Ｘはアミノ酸又はアミノ酸誘導体）がＣａＳＲアゴニスト活性を有する化合物であること等が記載されているが、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙについては、実施例において実際に合成・評価された旨の記載はなく、具体的に開示されていない。尚、特許文献１及び２の内容は、本明細書の記載に含まれるものとする。 Accordingly, many variation compounds having CaSR agonist activity can be searched for, and can have a more excellent body taste imparting action, in particular, a taste-type body taste imparting action, and can be easily produced at low cost with excellent stability. It is desired to find a substance capable of imparting a possible rich taste, to provide a rich taste imparting agent comprising the substance, and to provide a complex rich taste imparting agent comprising the substance in combination with another substance having CaSR agonist activity. It has been.
On the other hand, some γ-glutamyl peptides having γ-glutamine at the N-terminus are known to be synthesized as substrates in studies of enzyme activity or the like (Patent Document 3, Non-Patent Documents 1 and 2). There are no known examples where γ-Glu-Nva-Gly is actually used in food applications or naturally occurring.
Patent Document 1 describes that γ-Glu-X-Gly (X is an amino acid or an amino acid derivative) is a compound having CaSR agonist activity, etc., but γ-Glu-Nva-Gly is described. Is not described in detail in the examples, and is not disclosed specifically. In addition, the content of patent document 1 and 2 shall be contained in description of this specification.
国際公開第２００７／０５５３９３号パンフレットInternational Publication No. 2007/055393 Pamphlet 国際公開第２００８／１３９９４５号パンフレットInternational Publication No. 2008/139945 Pamphlet 国際公開第２００７／０６６４３０号パンフレットInternational Publication No. 2007/066430 Pamphlet
　本発明は、ＣａＳＲアゴニスト活性を有する多くのバリエーション化合物を探索してより優れたコク味付与作用、特に中後味型で高力価のコク味付与作用を有し、かつ安定性に優れた、コク味を付与するこのできる物質を見出し、該物質からなるコク味付与剤、及び該物質を他のＣａＳＲアゴニスト活性を有する物質と併用してなる複合コク味付与剤を提供することを課題とする。更に、一定濃度の該物質を含有する食品組成物を提供することを課題とする。 The present invention searches for many variation compounds having CaSR agonist activity and has a more excellent body taste imparting action, in particular, an aftertaste type and high potency body taste imparting action, and has excellent stability. It is an object of the present invention to find a substance capable of imparting a taste, and to provide a kokumi imparting agent comprising the substance and a complex kokumi imparting agent comprising the substance in combination with another substance having CaSR agonist activity. It is another object of the present invention to provide a food composition containing the substance at a certain concentration.
　本発明者は、種々の化合物を探索した結果、驚くべきことに、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ（Ｌ－γ－グルタミル－Ｌ－ノルバリル－グリシン）が、高いＣａＳＲアゴニスト活性と極めて優れたコク味付与効果を有し、特にその呈味パターンが中後味型であるコク味を付与することができることを見出した。さらに、見出されたγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙが、同様のトリペプチドであるγ－Ｇｌｕ－Ｖａｌ－Ｇｌｙと比較して10倍以上と、きわめて高力価であり、また安定性に優れ、かつより中後味が強いという好ましい呈味パターンを示すことを見出した。さらに、見出されたγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙが、単独で有用なコク味付与剤となりえることを見出した。また、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを添加することにより、コク味の増強した、好ましい食品組成物が得られることを見出した。さらに、該物質を他のＣａＳＲアゴニスト活性を有する物質と併用してなる複合コク味付与剤となりうることを見出し、本発明を完成させた。 As a result of searching for various compounds, the present inventor has surprisingly found that γ-Glu-Nva-Gly (L-γ-glutamyl-L-norvalyl-glycine) has high CaSR agonist activity and extremely excellent body taste. It has been found that it has an imparting effect, and in particular, its taste pattern can impart a rich taste that is a medium aftertaste type. Furthermore, the found γ-Glu-Nva-Gly has an extremely high titer of 10 times or more compared with the same tripeptide γ-Glu-Val-Gly, and has excellent stability. And it discovered that the preferable taste pattern that a middle aftertaste was strong was shown. Furthermore, it has been found that the found γ-Glu-Nva-Gly can be a useful body taste imparting agent by itself. Further, it has been found that by adding γ-Glu-Nva-Gly, a preferable food composition with enhanced richness can be obtained. Furthermore, it discovered that it could become a complex body taste imparting agent formed by using this substance together with another substance having CaSR agonist activity, and completed the present invention.
　すなわち、本発明は、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなるコク味付与剤を提供する。
　また、本発明はγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを含有する食品組成物をも提供する（以下、「本発明の食品組成物」ともいう。）。また、本発明は、（ａ）γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙに、（ｂ）γ－Ｇｌｕ－Ｘ－Ｇｌｙ（Ｘはアミノ酸又はアミノ酸誘導体を表す、但しＮｖａを除く）、γ－Ｇｌｕ－Ｖａｌ－Ｙ（Ｙはアミノ酸又はアミノ酸誘導体を表す）、γ－Ｇｌｕ－Ｎｖａ、γ－Ｇｌｕ－Ａｂｕ、γ－Ｇｌｕ－Ａｌａ、γ－Ｇｌｕ－Ｇｌｙ、γ－Ｇｌｕ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ、γ－Ｇｌｕ－Ｔｈｒ、γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｏｒｎ、Ａｓｐ－Ｇｌｙ、Ｃｙｓ－Ｇｌｙ、Ｃｙｓ－Ｍｅｔ、Ｇｌｕ－Ｃｙｓ、Ｇｌｙ－Ｃｙｓ、Ｌｅｕ－Ａｓｐ、Ｄ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ（Ｏ）、γ－Ｇｌｕ－γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｖａｌ－ＮＨ2、γ－Ｇｌｕ－Ｖａｌ－ｏｌ、γ－Ｇｌｕ－Ｓｅｒ、γ－Ｇｌｕ－Ｔａｕ、γ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）（Ｏ）、γ－Ｇｌｕ－Ｌｅｕ、γ－Ｇｌｕ－Ｉｌｅ、γ－Ｇｌｕ－ｔ－Ｌｅｕおよびγ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）からなる群より選択される１種又は２種以上のアミノ酸又はペプチド、を併用してなる複合コク味付与剤を提供する。 That is, the present invention provides a rich taste imparting agent comprising γ-Glu-Nva-Gly.
The present invention also provides a food composition containing γ-Glu-Nva-Gly (hereinafter also referred to as “the food composition of the present invention”). The present invention also relates to (a) γ-Glu-Nva-Gly, (b) γ-Glu-X-Gly (X represents an amino acid or amino acid derivative, except Nva), γ-Glu-Val- Y (Y represents an amino acid or amino acid derivative), γ-Glu-Nva, γ-Glu-Abu, γ-Glu-Ala, γ-Glu-Gly, γ-Glu-Cys, γ-Glu-Met, γ- Glu-Thr, γ-Glu-Val, γ-Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, γ-Glu-Met ( O), γ-Glu-γ-Glu-Val, γ-Glu-Val-NH 2 , γ-Glu-Val-ol, γ-Glu-Ser, γ-Glu-Tau, γ-Glu-Cys (S- Me) (O) One or more amino acids or peptides selected from the group consisting of γ-Glu-Leu, γ-Glu-Ile, γ-Glu-t-Leu and γ-Glu-Cys (S-Me), Provided is a complex rich taste imparting agent used in combination.
　本発明によれば、極めて優れたコク味付与作用、特に呈味パターンが図１に示されるようなプロフィールを有するユニークな中後味型の極めて強力なコク味付与作用を有し、かつ安定性に優れ簡便かつ低コストで生産することが可能なコク味付与剤、及び複合コク味付与剤を提供することができる。又、本発明によれば、優れたコク味付与作用を有する物質を一定濃度含有する、優れた食品組成物を提供することができる。
　本発明のコク味付与剤を用いると、低脂肪食品の呈味に脂肪様濃厚感及び滑らかさを付与できるので、脂肪含有食品中の脂肪の含有量を低下させても、元の食品と同様の濃厚感を保持でき、健康志向の高い食品にすることができる。このような食品としては、肉類含有食品や乳製品などがあげられる。特に、本発明のコク味付与剤を含有する食品を喫食すると、食べたとたんではなくて、その後に、脂肪様濃厚感及び滑らかさを感じることができるという利点がある。 According to the present invention, it has a very excellent body taste imparting action, in particular, a unique medium aftertaste type having a profile whose taste pattern is shown in FIG. A rich body taste imparting agent and a complex body taste imparting agent that can be produced easily and at low cost can be provided. Moreover, according to this invention, the outstanding food composition which contains the substance which has the outstanding richness imparting effect | action in a fixed density | concentration can be provided.
Using the rich taste imparting agent of the present invention can impart a fat-like richness and smoothness to the taste of low-fat foods, so even if the fat content in fat-containing foods is reduced, it is the same as the original food Can maintain a rich sense of health and can be made into a health-conscious food. Examples of such foods include meat-containing foods and dairy products. In particular, when a food containing the richness-imparting agent of the present invention is eaten, there is an advantage that a fat-like richness and smoothness can be felt afterwards without being eaten.
図１は、中後味型のコク味付与剤の味覚プロフィールを示す。FIG. 1 shows a taste profile of a medium aftertaste type body taste imparting agent.
　本発明で対象とするγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは３つのアミノ酸がペプチド結合してなるＬ－γ－グルタミル－Ｌ－ノルバリル－グリシン及び/又はその塩、特に可食性の塩が含まれる。
　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは優れたコク味付与効果を有するため、コク味付与剤として用いることができる。γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは、コク味を付与する食品組成物の重量に対して、０．１ｐｐｂ～９９．９質量％、好ましくは１ｐｐｂ～１０質量％、より好ましくは０．０１ｐｐｍ～１質量％、含有するように添加して用いることができる。すなわち、本発明の別の態様は、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを含有する食品組成物、好ましくは、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを０．１ｐｐｂ～９９．９質量％含有する食品組成物に関する。より好ましくは、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを０．０１～５０重量ｐｐｍ含有する食品組成物に関する。 Γ-Glu-Nva-Gly targeted in the present invention includes L-γ-glutamyl-L-norvalyl-glycine formed by peptide bonding of three amino acids and / or a salt thereof, particularly an edible salt.
Since γ-Glu-Nva-Gly has an excellent body taste imparting effect, it can be used as a body taste imparting agent. γ-Glu-Nva-Gly is 0.1 ppb to 99.9% by mass, preferably 1 ppb to 10% by mass, more preferably 0.01 ppm to 1% by mass with respect to the weight of the food composition imparting a rich taste. % Can be added and used. That is, another aspect of the present invention relates to a food composition containing γ-Glu-Nva-Gly, preferably a food composition containing 0.1 ppb to 99.9% by mass of γ-Glu-Nva-Gly. . More preferably, the present invention relates to a food composition containing 0.01 to 50 ppm by weight of γ-Glu-Nva-Gly.
　また、本発明のコク味付与剤、すなわち、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは、グルタミン酸ナトリウム（ＭＳＧ）などのアミノ酸類、イノシン一リン酸（ＩＭＰ）などの核酸類、塩化ナトリウムなどの無機塩類、クエン酸などの有機酸類、種々の酵母エキスなどから選択される少なくとも１種の他の調味原料と組み合わせて用いることにより、他の調味原料を単独で用いる場合に比べて、よりコク味の増した、好ましい調味料を提供することができる。 Further, the body taste imparting agent of the present invention, that is, γ-Glu-Nva-Gly is composed of amino acids such as sodium glutamate (MSG), nucleic acids such as inosine monophosphate (IMP), inorganic salts such as sodium chloride, By using in combination with at least one other seasoning raw material selected from organic acids such as citric acid, various yeast extracts, etc., the richness of the flavor increased more than when using other seasoning raw materials alone. A preferable seasoning can be provided.
　本発明において「コク味」とは、甘味（sweet taste）、塩味（salty taste）、酸味（sour taste）、苦味（bitter taste）、うま味（umami）で表される５基本味（five basic tastes）では表すことのできない味を意味し、基本味だけでなく、厚み（thickness）・ひろがり（growth（mounthfulness））・持続性（continuity）・まとまり（harmony）など基本味の周辺の味（marginal tastes）をも増強した味をいう。ここで、「コク味付与」とは、甘味、塩味、酸味、苦味、うま味で表される５基本味の増強と、それに伴う厚み・ひろがり・まとまりなど基本味の周辺の味を付与することをいう。また、これを呈味増強作用と表現することもできる。したがって、本発明のコク味付与剤であるγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは、呈味増強剤（Flavor Enhancer）と表現することもできる。本発明のコク味付与剤であるγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは、甘味増強剤、塩味増強剤、酸味増強剤、苦味増強剤またはうま味増強剤として使用することができる。
　また、味覚は喫食後の時間経過とともに変化するが、喫食直後から順に、先味（initial taste）、中味（middle taste）及び後味（after taste）と呼ぶ。これらは相対的な概念であるが、概して、先味、中味及び後味は、それぞれ喫食後０から２秒まで、２秒から５秒まで、及び５秒以降に感じる呈味である。また、０から５秒までを「先中味」といい、２秒以降約３０秒前後までを「中後味」とする（図１参照）。３区分に分けた評価について、喫食者の評価への集中が困難なため、通常２区分に分けた評価を常用する。
　コク味及び呈味パターンに対するＣａＳＲ活性を有する物質の効果は、ヒトによる味覚試験などの方法によって確認することができる。このようなヒトによる味覚官能試験としては、例えば本願明細書の実施例で示される試験が挙げられるが、これらに限定されない。 In the present invention, the term “kokumi” means five basic tastes represented by sweet taste, salty taste, sour taste, bitter taste, and umami. It means a taste that cannot be expressed in terms of basic taste, as well as thickness, thickness (mounthfulness), continuity, harmony, etc. The taste is also enhanced. Here, “kokumi impartation” means to enhance the five basic tastes expressed by sweetness, salty taste, acidity, bitterness, umami, and to give a taste around the basic tastes such as thickness, spread, and unity. Say. This can also be expressed as a taste enhancing action. Therefore, γ-Glu-Nva-Gly, which is the body taste imparting agent of the present invention, can also be expressed as a flavor enhancer. Γ-Glu-Nva-Gly which is a body taste imparting agent of the present invention can be used as a sweetness enhancer, salty taste enhancer, sour taste enhancer, bitterness enhancer or umami enhancer.
In addition, the taste changes with the passage of time after eating, but they are called an initial taste, a middle taste, and an after taste in order from immediately after eating. These are relative concepts, but in general, taste, medium and aftertaste are tastes to be felt from 0 to 2 seconds, from 2 to 5 seconds, and after 5 seconds, respectively, after eating. Further, the period from 0 to 5 seconds is called “first taste”, and the period from about 2 seconds to about 30 seconds is called “medium after taste” (see FIG. 1). Regarding the evaluation divided into three categories, it is difficult to concentrate on the evaluation of the eater, so the evaluation divided into two categories is usually used.
The effect of a substance having CaSR activity on kokumi and taste patterns can be confirmed by methods such as a human taste test. Examples of such human taste sensory tests include, but are not limited to, the tests shown in the Examples of the present specification.
　本明細書において、「ＣａＳＲ」とは、カルシウムセンシング受容体（Calcium Sensing Receptor）を意味し、７回膜貫通型受容体のクラスＣに属するものであり、カルシウム受容体とも呼ばれる。本明細書において「ＣａＳＲアゴニスト」とは、上記ＣａＳＲに結合し、ＣａＳＲを活性化する物質を意味する。また、本明細書において、「ＣａＳＲを活性化する」とは、ＣａＳＲにリガンドが結合し、グアニンヌクレオチド結合タンパク質を活性化して、シグナルを伝達することを意味する。また、ＣａＳＲに結合し、ＣａＳＲを活性化する性質を「ＣａＳＲアゴニスト活性」という。 In this specification, “CaSR” means a calcium sensing receptor (Calcium® Sensing® Receptor), which belongs to the class C of the 7-transmembrane receptor, and is also referred to as a calcium receptor. In the present specification, the “CaSR agonist” means a substance that binds to the CaSR and activates the CaSR. In the present specification, “activate CaSR” means that a ligand binds to CaSR and activates a guanine nucleotide-binding protein to transmit a signal. The property of binding to CaSR and activating CaSR is referred to as “CaSR agonist activity”.
　ＣａＳＲアゴニスト活性を有する化合物をスクリーニングする方法を具体的に示すが、これらのステップに限定されるものではない。
１）ＣａＳＲ活性を測定するためのＣａＳＲ活性測定系に被検物質を添加して、ＣａＳＲ活性を測定する。
２）被検物質を添加したときのＣａＳＲ活性と、被検物質を添加しなかったときのＣａＳＲ活性を比較する。
３）被検物質を添加したときにＣａＳＲアゴニスト活性を示す被検物質を選択する。 Although a method for screening a compound having CaSR agonist activity is specifically shown, it is not limited to these steps.
1) A test substance is added to a CaSR activity measurement system for measuring CaSR activity, and the CaSR activity is measured.
2) The CaSR activity when the test substance is added is compared with the CaSR activity when the test substance is not added.
3) Select a test substance that exhibits CaSR agonist activity when the test substance is added.
　ＣａＳＲ活性の測定は、例えば、ＣａＳＲを発現する細胞を用いた測定系を用いて行うことができる。上記細胞は、ＣａＳＲを内在的に発現する細胞であっても、外来的にＣａＳＲ遺伝子を導入した組み換え細胞であってもよい。上記ＣａＳＲ活性測定系は、上記ＣａＳＲを発現する細胞に、ＣａＳＲに特異的な細胞外リガンド（活性化物質）を加えたときに、活性化物質とＣａＳＲとの結合（反応）を検出することができるか、又は、活性化物質とＣａＳＲとの結合（反応）に応答して細胞内に検出可能なシグナルを伝達するものであれば、特に制限なく用いることができる。被検物質との反応によりＣａＳＲ活性が検出された場合、当該被検物質はＣａＳＲ刺激活性を有すると判定される。 The measurement of CaSR activity can be performed using, for example, a measurement system using cells that express CaSR. The cell may be a cell that endogenously expresses CaSR or a recombinant cell into which a CaSR gene has been introduced exogenously. The CaSR activity measurement system can detect binding (reaction) between an activator and CaSR when an extracellular ligand (activator) specific to CaSR is added to the cell expressing CaSR. It can be used without particular limitation as long as it can transmit a detectable signal in the cell in response to the binding (reaction) between the activator and CaSR. When CaSR activity is detected by reaction with the test substance, it is determined that the test substance has CaSR stimulating activity.
　上記ＣａＳＲとしては、GenBank Accession No. NM_000388で登録されているヒトＣａＳＲ遺伝子によってコードされるヒトＣａＳＲが好ましく例示できる。尚、ＣａＳＲは、上記配列の遺伝子によってコードされるタンパク質に制限されず、ＣａＳＲ機能を有するタンパク質をコードする限りにおいて、上記配列と６０％以上、好ましくは８０％以上、より好ましくは９０％以上の相同性を有する遺伝子によってコードされるタンパク質であってもよい。なお、ＣａＳＲ機能はこれらの遺伝子を細胞に発現させ、カルシウム添加時の電流の変化や細胞内カルシウムイオン濃度の変化を測定することによって調べることができる。
　上記ＣａＳＲは、その由来は特に制限されず、上記ヒトのＣａＳＲのみならず、マウス、ラット、イヌなどを含むあらゆる動物由来のＣａＳＲが挙げられる。 Preferred examples of the CaSR include human CaSR encoded by the human CaSR gene registered under GenBank Accession No. NM_000388. CaSR is not limited to the protein encoded by the gene of the above sequence, and as long as it encodes a protein having a CaSR function, it is 60% or more, preferably 80% or more, more preferably 90% or more. It may be a protein encoded by a homologous gene. The CaSR function can be examined by expressing these genes in cells and measuring changes in current and intracellular calcium ion concentration when calcium is added.
The origin of the CaSR is not particularly limited, and examples include CaSR derived from any animal including mice, rats, dogs and the like as well as the human CaSR.
　上述の如く、ＣａＳＲ活性は、ＣａＳＲ又はその断片を発現した生きた細胞、ＣａＳＲ又はその断片を発現した細胞膜、ＣａＳＲ又はその断片のタンパク質を含むインビトロの系などを利用して確認することができる。
　以下に生きた細胞を用いた一例を示すが、これに限定されるものではない。
　ＣａＳＲは、アフリカツメガエル卵母細胞やハムスター卵巣細胞やヒト胎児腎臓細胞等の培養細胞に発現させる。これは外来遺伝子を保持するプラスミドにＣａＳＲ遺伝子をクリーニングしたものを、プラスミドの状態もしくはそれを鋳型にしたｃＲＮＡを導入することで可能となる。反応の検出には電気生理学的手法や細胞内カルシウム上昇の蛍光指示試薬を用いることができる。
　ＣａＳＲの発現は、初めにカルシウムもしくは特異的活性化剤による応答で確認する。５ｍＭ程度の濃度のカルシウムに対して、細胞内電流が観察された卵母細胞もしくは蛍光指示試薬の蛍光が観察された培養細胞を使用する。カルシウムの濃度を変えて濃度依存性を測定する。次に、被検物質を１μＭ～１ｍＭ程度に調製し、卵母細胞もしくは培養細胞に添加し、上記被検物質存在下でのＣａＳＲ活性を測定することで、上記被検物質のＣａＳＲアゴニスト活性を測定する。
　又、より具体的には、ＣａＳＲアゴニスト活性試験としては例えば本願明細書の試験例で示される試験が挙げられるが、これらに限定されない。 As described above, the CaSR activity can be confirmed using a living cell expressing CaSR or a fragment thereof, a cell membrane expressing CaSR or a fragment thereof, an in vitro system containing a protein of CaSR or a fragment thereof, or the like.
An example using living cells is shown below, but is not limited thereto.
CaSR is expressed in cultured cells such as Xenopus oocytes, hamster ovary cells, and human fetal kidney cells. This can be achieved by introducing a plasmid carrying a foreign gene into which a CaSR gene has been cleaned and introducing the plasmid state or cRNA using it as a template. For the detection of the reaction, an electrophysiological technique or a fluorescent indicator reagent for increasing intracellular calcium can be used.
CaSR expression is first confirmed by a response with calcium or a specific activator. An oocyte in which an intracellular current is observed or a cultured cell in which fluorescence of a fluorescent indicator reagent is observed is used with respect to calcium having a concentration of about 5 mM. The concentration dependence is measured by changing the calcium concentration. Next, the test substance is prepared to about 1 μM to 1 mM, added to the oocyte or cultured cell, and the CaSR activity of the test substance is measured by measuring the CaSR activity in the presence of the test substance. taking measurement.
More specifically, examples of the CaSR agonist activity test include, but are not limited to, the tests shown in the test examples of the present specification.
　本発明の複合コク味付与剤においてγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙと併用されるアミノ酸又はペプチドは、γ－Ｇｌｕ－Ｘ－Ｇｌｙ（ＸはＮｖａ以外のアミノ酸又はアミノ酸誘導体を表す）、γ－Ｇｌｕ－Ｖａｌ－Ｙ（Ｙはアミノ酸又はアミノ酸誘導体を表す）、γ－Ｇｌｕ－Ｎｖａ、γ－Ｇｌｕ－Ａｂｕ、γ－Ｇｌｕ－Ａｌａ、γ－Ｇｌｕ－Ｇｌｙ、γ－Ｇｌｕ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ、γ－Ｇｌｕ－Ｔｈｒ、γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｏｒｎ、Ａｓｐ－Ｇｌｙ、Ｃｙｓ－Ｇｌｙ、Ｃｙｓ－Ｍｅｔ、Ｇｌｕ－Ｃｙｓ、Ｇｌｙ－Ｃｙｓ、Ｌｅｕ－Ａｓｐ、Ｄ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ（Ｏ）、γ－Ｇｌｕ－γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｖａｌ－ＮＨ2、γ－Ｇｌｕ－Ｖａｌ－ｏｌ、γ－Ｇｌｕ－Ｓｅｒ、γ－Ｇｌｕ－Ｔａｕ、γ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）（Ｏ）、γ－Ｇｌｕ－Ｌｅｕ、γ－Ｇｌｕ－Ｉｌｅ、γ－Ｇｌｕ－ｔ－Ｌｅｕおよびγ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）からなる群より選択される１種又は２種以上のアミノ酸又はペプチドであるが、ここで、アミノ酸とは、Ｇｌｙ、Ａｌａ、Ｖａｌ、Ｌｅｕ、Ｉｌｅ、Ｓｅｒ、Ｔｈｒ、Ｃｙｓ、Ｍｅｔ、Ａｓｎ、Ｇｌｎ、Ｐｒｏ、Ｈｙｐ、ｔ－Ｌｅｕ等の中性アミノ酸、Ａｓｐ、Ｇｌｕ等の酸性アミノ酸、Ｌｙｓ、Ａｒｇ、Ｈｉｓ等の塩基性アミノ酸、Ｐｈｅ、Ｔｙｒ、Ｔｒｐ等の芳香族アミノ酸や、ホモセリン、シトルリン、オルニチン、α－アミノ酪酸、ノルバリン、ノルロイシン、タウリン等も含まれる。また、ｔｅｒｔ－ロイシン、シクロロインシン、α－アミノイソブチル酸、Ｌ－ペニシラミン等の非天然（非タンパク質構成）アミノ酸であってもよい。尚、ペプチドγ－Ｇｌｕ－Ｘ－Ｇｌｙにおいては、Ｘは上記のようなアミノ酸又はその誘導体のいずれでもよいが、Ｃｙｓ以外のアミノ酸又はその誘導体が好ましい。中でも、併用されるペプチドとしては、γ－Ｇｌｕ－Ｖａｌ－Ｇｌｙ、γ－Ｇｌｕ－Ａｂｕ－Ｇｌｙ、γ－Ｇｌｕ－ｔＬｅｕ－Ｇｌｙ、γ－Ｇｌｕ－Ｎｖａおよびγ－Ｇｌｕ－Ａｂｕ等が好ましい。
　特に、本発明のコク味付与剤はγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなり、図１に示されるようなプロフィールを有するユニークな中後味型の優れたコク味付与作用を有するので、このようなプロフィールとは異なるプロフィールを有するペプチト、例えば、先味傾向のγ－Ｇｌｕ－Ａｂｕ－Ｇｌｙ、γ－Ｇｌｕ－Ａｂｕなどと組み合わせて用いるのが好ましい。 The amino acid or peptide used in combination with γ-Glu-Nva-Gly in the complex rich taste imparting agent of the present invention is γ-Glu-X-Gly (X represents an amino acid or amino acid derivative other than Nva), γ-Glu- Val-Y (Y represents an amino acid or amino acid derivative), γ-Glu-Nva, γ-Glu-Abu, γ-Glu-Ala, γ-Glu-Gly, γ-Glu-Cys, γ-Glu-Met, γ-Glu-Thr, γ-Glu-Val, γ-Glu-Orn, Asp-Gly, Cys-Gly, Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, γ-Glu- Met (O), γ-Glu-γ-Glu-Val, γ-Glu-Val-NH 2 , γ-Glu-Val-ol, γ-Glu-Ser, γ-Glu-Tau, γ-G 1 selected from the group consisting of lu-Cys (S-Me) (O), γ-Glu-Leu, γ-Glu-Ile, γ-Glu-t-Leu and γ-Glu-Cys (S-Me) Species or two or more amino acids or peptides, where amino acids are Gly, Ala, Val, Leu, Ile, Ser, Thr, Cys, Met, Asn, Gln, Pro, Hyp, t-Leu, etc. Neutral amino acids, acidic amino acids such as Asp and Glu, basic amino acids such as Lys, Arg and His, aromatic amino acids such as Phe, Tyr and Trp, homoserine, citrulline, ornithine, α-aminobutyric acid, norvaline, norleucine , Taurine and the like are also included. Further, it may be a non-natural (non-protein constituent) amino acid such as tert-leucine, cycloloinsine, α-aminoisobutyric acid, L-penicillamine and the like. In the peptide γ-Glu-X-Gly, X may be any of the above amino acids or derivatives thereof, but an amino acid other than Cys or a derivative thereof is preferred. Among these, γ-Glu-Val-Gly, γ-Glu-Abu-Gly, γ-Glu-tLeu-Gly, γ-Glu-Nva, γ-Glu-Abu and the like are preferable.
In particular, the body taste imparting agent of the present invention is composed of γ-Glu-Nva-Gly and has a unique medium aftertaste type excellent body taste imparting action having a profile as shown in FIG. It is preferably used in combination with a peptide having a different profile from γ-Glu-Abu-Gly, γ-Glu-Abu, etc.
　本明細書において、アミノ酸残基の略号は以下のアミノ酸を意味する。
（１）Ｇｌｙ：グリシン
（２）Ａｌａ：アラニン
（３）Ｖａｌ：バリン
（４）Ｌｅｕ：ロイシン
（５）Ｉｌｅ：イソロイシン
（６）Ｍｅｔ：メチオニン
（７）Ｐｈｅ：フェニルアラニン
（８）Ｔｙｒ：チロシン
（９）Ｔｒｐ：トリプトファン
（10）Ｈｉｓ：ヒスチジン
（11）Ｌｙｓ：リジン
（12）Ａｒｇ：アルギニン
（13）Ｓｅｒ：セリン
（14）Ｔｈｒ：トレオニン
（15）Ａｓｐ：アスパラギン酸
（16）Ｇｌｕ：グルタミン酸
（17）Ａｓｎ：アルパラギン
（18）Ｇｌｎ：グルタミン
（19）Ｃｙｓ：システイン
（20）Ｐｒｏ：プロリン
（21）Ｏｒｎ：オルニチン
（22）Ｓａｒ：サルコシン
（23）Ｃｉｔ：シトルリン
（24）Ｎ－Ｖａｌ（又は、Ｎｖａ）：ノルバリン　（２－アミノ吉草酸）
（25）Ｎ－Ｌｅｕ（又は、Ｎｌｅ）：ノルロイシン
（26）Ａｂｕ：α－アミノ酪酸
（27）Ｔａｕ：タウリン
（28）Ｈｙｐ：ヒドロキシプロリン
（29）ｔ－Ｌｅｕ：ｔｅｒｔ－ロイシン
（30）Ｃｌｅ：シクロロイシン
（31）Ａｉｂ：α－アミノイソブチル酸（α－aminoisobutyric acid、２－メチルアラニン）
（32）Ｐｅｎ：Ｌ－ペニシラミン（penicillamine）
（33）ａｌｌｏ－Ｔｈｒ：アロスレオニン
（34）ａｌｌｏ－Ｉｌｅ：アロイソロイシン In this specification, the abbreviation of an amino acid residue means the following amino acids.
(1) Gly: glycine (2) Ala: alanine (3) Val: valine (4) Leu: leucine (5) Ile: isoleucine (6) Met: methionine (7) Phe: phenylalanine (8) Tyr: tyrosine (9 ) Trp: tryptophan (10) His: histidine (11) Lys: lysine (12) Arg: arginine (13) Ser: serine (14) Thr: threonine (15) Asp: aspartic acid (16) Glu: glutamic acid (17) Asn: Asparagine (18) Gln: Glutamine (19) Cys: Cysteine (20) Pro: Proline (21) Orn: Ornithine (22) Sar: Sarcosine (23) Cit: Citrulline (24) N-Val (or Nva) : Norvaline (2-aminovaleric acid)
(25) N-Leu (or Nle): norleucine (26) Abu: α-aminobutyric acid (27) Tau: taurine (28) Hyp: hydroxyproline (29) t-Leu: tert-leucine (30) Cle: Cycloleucine (31) Aib: α-aminoisobutyric acid (2-methylalanine)
(32) Pen: L-penicillamine
(33) allo-Thr: arosleonine (34) allo-Ile: alloisoleucine
　また、アミノ酸誘導体とは、上記アミノ酸の各種誘導体であって、例えば、特殊アミノ酸や非天然アミノ酸、アミノアルコール、或いは末端カルボニル基やアミノ基、システインのチオール基等のアミノ酸側鎖が各種置換基により置換したものが挙げられる。置換基としては、アルキル基、アシル基、水酸基、アミノ基、アルキルアミノ基、ニトロ基、スルホニル基や各種保護基等が挙げられ、例えば、Ａｒｇ（ＮＯ2）：Ｎ－γ－ニトロアルギニン、Ｃｙｓ（ＳＮＯ）：Ｓ－ニトロシステイン、Ｃｙｓ（Ｓ－Ｍｅ）：Ｓ－メチルシステイン、Ｃｙｓ（Ｓ－allyl）：Ｓ－アリルシステイン、Ｖａｌ－ＮＨ2：バリンアミド、Ｖａｌ－ｏｌ：バリノール（２－アミノ－３－メチル－１－ブタノール）等が含まれる。尚、本明細書において、γ－Ｇｌｕ－Ｃｙｓ（ＳＮＯ）－Ｇｌｙは下記の構造式を有するものであり、上記γ－Ｇｌｕ－Ｍｅｔ（Ｏ）およびγ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）（Ｏ）式中の（Ｏ）はスルホキシド構造であることを意味する。γ－Ｇｌｕの（γ）とは、グルタミン酸のγ位のカルボキシル基を介して他のアミノ酸が結合していることを意味する。
Figure JPOXMLDOC01-appb-C000001
 The amino acid derivatives are various derivatives of the above amino acids. For example, amino acids side chains such as special amino acids, unnatural amino acids, amino alcohols, terminal carbonyl groups, amino groups, cysteine thiol groups, and the like are substituted with various substituents. Substituted ones are mentioned. Examples of the substituent include an alkyl group, an acyl group, a hydroxyl group, an amino group, an alkylamino group, a nitro group, a sulfonyl group, and various protective groups. For example, Arg (NO 2 ): N-γ-nitroarginine, Cys (SNO): S-nitrocysteine, Cys (S-Me): S-methylcysteine, Cys (S-allyl): S-allylcysteine, Val-NH 2 : Valinamide, Val-ol: Valinol (2-amino- 3-methyl-1-butanol) and the like. In the present specification, γ-Glu-Cys (SNO) -Gly has the following structural formula, and γ-Glu-Met (O) and γ-Glu-Cys (S-Me) (O (O) in the formula means a sulfoxide structure. (γ) of γ-Glu means that another amino acid is bonded via a carboxyl group at the γ position of glutamic acid.
Figure JPOXMLDOC01-appb-C000001
　本発明のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ及びこれと併用されるアミノ酸又はペプチドは、市販されている場合には市販品を用いることもでき、その他、（１）化学的に合成する方法、又は（２）酵素的な反応により合成する方法等の公知手法を適宜用いることによって取得することができるが、化学的な合成がより簡便である。本発明において用いられるγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙは含まれるアミノ酸の残基数が３残基と短いので、化学的に合成する方法が比較的簡便であり、産業上有利である。また、本発明のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ及びこれと併用されるアミノ酸又はペプチドを化学的に合成する場合には、該オリゴペプチドをペプチド合成機を用いて合成あるいは半合成することにより行うことができる。化学的に合成する方法としては、例えばペプチド固相合成法等が挙げられる。そのようにして合成したペプチドは通常の手段、例えばイオン交換クロマトグラフィー、逆相高速液体クロマトグラフィー、アフィニティークロマトグラフィー等によって精製することができる。このようなペプチド固相合成法、およびそれに続きペプチド精製はこの技術分野においてよく知られたものである。 The γ-Glu-Nva-Gly and the amino acid or peptide used in combination with the γ-Glu-Nva-Gly of the present invention may be a commercially available product, in addition, (1) a chemical synthesis method, or ( 2) Although it can be obtained by appropriately using a known method such as a method of synthesizing by an enzymatic reaction, chemical synthesis is simpler. Since γ-Glu-Nva-Gly used in the present invention has a short amino acid residue of 3 residues, the chemical synthesis method is relatively simple and industrially advantageous. In addition, when the γ-Glu-Nva-Gly of the present invention and the amino acid or peptide used in combination therewith are chemically synthesized, the oligopeptide is synthesized or semi-synthesized using a peptide synthesizer. Can do. Examples of the chemical synthesis method include a peptide solid phase synthesis method. The peptide thus synthesized can be purified by conventional means such as ion exchange chromatography, reverse phase high performance liquid chromatography, affinity chromatography and the like. Such peptide solid phase synthesis methods, and subsequent peptide purification, are well known in the art.
　また、本発明において用いられるγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ及びこれと併用されるアミノ酸又はペプチドを酵素的な反応により生産する場合には、例えば、国際公開パンフレットＷＯ２００４／０１１６５３号に記載の方法を用いてもよい。すなわち、一方のアミノ酸又はジペプチドのカルボキシル末端をエステル化又はアミド化したアミノ酸又はジペプチドと、アミノ酸がフリーの状態であるアミノ酸（例えば、カルボキシル基が保護されたアミノ酸）とを、ペプチド生成酵素の存在下において反応させ、生成したジペプチド又はトリペプチドを生成することによっても生産することが可能である。ペプチド生成酵素としては、ペプチドを生成する能力を有する微生物の培養物、該培養物より分離した微生物菌体、又は該微生物の菌体処理物、又は該微生物に由来するペプチド生成酵素が挙げられる。尚、ＷＯ２００４／０１１６５３号に記載されている事項は、本明細書の記載に含まれるものとする。
　さらに、上述したような酵素的な方法や化学的合成方法以外にも本発明において用いられるペプチドが、野菜や果物等の植物、酵母等の微生物、その他の天然物中に存在する場合がある。天然に存在する場合には、これらから抽出して用いることも可能である。
　本発明のコク味付与剤あるいは複合コク味付与剤は、そのままで、又は飲食品的に許容しうる担体や他の調味原料と混合して、調味料とすることができる。他の調味原料としては、例えば、香料、糖類、甘味料、食物繊維類、ビタミン類、グルタミン酸ナトリウム（ＭＳＧ）などのアミノ酸類、イノシン一リン酸（ＩＭＰ）などの核酸類、塩化ナトリウムなどの無機塩類、クエン酸などの有機酸類が挙げられ、更には種々の酵母エキスも挙げられる。 In the case of producing γ-Glu-Nva-Gly and an amino acid or peptide used in combination therewith by an enzymatic reaction, for example, the method described in International Publication Pamphlet WO 2004/011653 is used. May be. That is, an amino acid or dipeptide in which the carboxyl terminus of one amino acid or dipeptide is esterified or amidated, and an amino acid in which the amino acid is free (for example, an amino acid in which the carboxyl group is protected) are combined in the presence of a peptide-forming enzyme. It is also possible to produce by producing the dipeptide or tripeptide produced by reacting in the above. Examples of the peptide-forming enzyme include a culture of a microorganism having the ability to produce a peptide, a microbial cell separated from the culture, a treated product of the microorganism, or a peptide-generating enzyme derived from the microorganism. The matters described in WO 2004/011653 are included in the description of this specification.
Furthermore, in addition to the enzymatic methods and chemical synthesis methods described above, the peptides used in the present invention may be present in plants such as vegetables and fruits, microorganisms such as yeast, and other natural products. If they exist in nature, they can be extracted from these and used.
The kokumi imparting agent or the complex kokumi imparting agent of the present invention can be used as a seasoning as it is or by mixing it with a carrier and other seasoning ingredients that are acceptable for food and drink. Other seasoning materials include, for example, flavorings, sugars, sweeteners, dietary fibers, vitamins, amino acids such as sodium glutamate (MSG), nucleic acids such as inosine monophosphate (IMP), and inorganic substances such as sodium chloride. Examples thereof include organic acids such as salts and citric acid, and various yeast extracts.
　尚、本発明のコク味付与剤あるいは複合コク味付与剤を含有する食品組成物として好ましい低脂肪食品は、元来脂肪を含む食品であり、特に、その脂肪含量が低減された食品である。ここで、「脂肪」とは「油脂」と同義であり、固形と液状の両方を含み、動物性脂肪及び植物性脂肪のいずれであってもよい。
　このような低脂肪食品としては、牛乳、ヨーグルト、バター、クリーム等の乳製品、マーガリン、コーヒー用ミルク、 ソース、ルーなどの動物油脂及び/又は植物油脂含有食品、ドレッシング、マヨネーズなどの乳化食品等、調理済みの肉類を含む各種カレーやシチュー、肉エキスを含む各種スープなどがあげられる。又、調理済みの低脂肪牛肉からなるステーキや焼き肉類などや通常のフライ処理ではないベイクドスナックもあげられる。これらのうち、低脂肪食品としては、通常食品の脂肪含有量が1/2～1/3であるものが好ましい。
　本発明のコク味付与剤を上記低脂肪食品に含有させることにより、これらの食品を喫食した時、最初ではなくてその次に、脂肪様濃厚感及び滑らかさを感じることができる。
　なお、牛乳やヨーグルトに関しては、通常品が脂肪３～４％であるのに対し、ゼロ脂肪品（脂肪０．１％前後）も知られている。本発明のコク味付与剤あるいは複合コク味付与剤はこれらゼロ脂肪品にも有効である。 In addition, the low fat food preferable as a food composition containing the rich taste imparting agent or the complex rich taste imparting agent of the present invention is a food originally containing fat, and particularly a food having a reduced fat content. Here, “fat” is synonymous with “oil and fat”, includes both solid and liquid, and may be either animal fat or vegetable fat.
Such low-fat foods include dairy products such as milk, yogurt, butter and cream, margarine, milk for coffee, foods containing animal oils and / or vegetable oils such as sauces, roux, emulsified foods such as dressings and mayonnaise, etc. And various curries and stews containing cooked meat, and various soups containing meat extracts. In addition, steaks and grilled meat made of cooked low-fat beef and baked snacks that are not subjected to normal frying are also included. Among these, as the low-fat food, those whose normal fat content is 1/2 to 1/3 are preferable.
By containing the rich taste-imparting agent of the present invention in the low-fat foods, when these foods are eaten, the fat-like richness and smoothness can be felt next, not first.
Regarding milk and yogurt, normal products are 3-4% fat, while zero fat products (about 0.1% fat) are also known. The savoriness imparting agent or the complex savoriness imparting agent of the present invention is also effective for these zero fat products.
　また、本発明のコク味付与剤あるいは複合コク味付与剤は、とりわけ、ポーク原料を含有する食品に添加することも好ましい。すなわち、本発明は、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙとポーク原料とを含有する食品組成物も提供する。ポーク原料を含有する食品としては、特に制限はないが、例えば、ポークエキス、ソーセージ、即麺スープ等が挙げられる。ポーク原料の含有量に特に制限はないが、例えば、本発明の食品組成物において、0.005～８０重量％程度であるものが挙げられる。
　また、本発明のコク味付与剤あるいは複合コク味付与剤は、ビーフ原料を含有する食品に添加することも好ましい。すなわち、本発明は、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙとビーフ原料とを含有する食品組成物も提供する。ビーフ原料を含有する食品としては、特に制限はないが、例えば、ビーフエキス、コーンビーフ、ビーフ使用スープ、ビーフ使用ソース等が挙げられる。ビーフ原料の含有量に特に制限はないが、例えば食品組成物において、0.005～８０重量％程度であるものが挙げられる。 Moreover, it is also preferable to add the rich taste imparting agent or the complex rich taste imparting agent of the present invention to foods containing pork raw materials, among others. That is, the present invention also provides a food composition containing γ-Glu-Nva-Gly and pork ingredients. The food containing the pork raw material is not particularly limited, and examples thereof include pork extract, sausage, and instant noodle soup. The content of the pork raw material is not particularly limited, and examples thereof include a food composition of the present invention that is about 0.005 to 80% by weight.
Moreover, it is also preferable to add the rich taste imparting agent or the complex rich taste imparting agent of the present invention to foods containing beef raw materials. That is, the present invention also provides a food composition containing γ-Glu-Nva-Gly and a beef raw material. Although there is no restriction | limiting in particular as a foodstuff containing a beef raw material, For example, beef extract, corn beef, beef use soup, beef use sauce, etc. are mentioned. There is no particular limitation on the content of the beef raw material, but for example, a food composition may be about 0.005 to 80% by weight.
　本発明において用いられるγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ及び併用されるアミノ酸又はペプチドは塩の形態をも包含する。本発明のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ及び併用されるアミノ酸又はペプチドが塩の形態を形成し得る場合、その塩は薬理学的に許容される、可食性の塩であればよく、例えば、カルボキシル基等の酸性基に対しては、アンモニウム塩、ナトリウム、カリウム等のアルカリ金属との塩、カルシウム、マグネシウム等のアルカリ土類金属との塩、アルミニウム塩、亜鉛塩、トリエチルアミン、エタノールアミン、モルホリン、ピロリジン、ピペリジン、ピペラジン、ジシクロヘキシルアミン等の有機アミンとの塩、アルギニン、リジン等の塩基性アミン酸との塩を挙げることができる。また、塩基性基に対しては、塩酸、硫酸、リン酸、硝酸、臭化水素酸等の無機酸との塩、酢酸、クエン酸、安息香酸、マレイン酸、フマル酸、酒石酸、コハク酸、タンニン酸、酪酸、ヒベンズ酸、パモ酸、エナント酸、デカン酸、テオクル酸、サリチル酸、乳酸、シュウ酸、マンデル酸、リンゴ酸等の有機カルボン酸との塩、メタンスルホン酸、ベンゼンスルホン酸、ｐ－トルエンスルホン酸等の有機スルホン酸との塩を挙げることができる。 Γ-Glu-Nva-Gly and the amino acid or peptide used together in the present invention also include a salt form. When the γ-Glu-Nva-Gly of the present invention and the amino acid or peptide used in combination can form a salt form, the salt may be a pharmacologically acceptable edible salt. For acidic groups such as groups, ammonium salts, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnesium, aluminum salts, zinc salts, triethylamine, ethanolamine, morpholine, Examples thereof include salts with organic amines such as pyrrolidine, piperidine, piperazine and dicyclohexylamine, and salts with basic amine acids such as arginine and lysine. For basic groups, salts with inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, hydrobromic acid, acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, succinic acid, Salts with organic carboxylic acids such as tannic acid, butyric acid, hibenzic acid, pamoic acid, enanthic acid, decanoic acid, teocric acid, salicylic acid, lactic acid, oxalic acid, mandelic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p And salts with organic sulfonic acids such as toluenesulfonic acid.
　本発明のコク味付与剤、食品組成物、あるいは複合コク味付与剤は、乾燥粉末、ペースト、溶液などの物性に制限なしにあらゆる形態で用いることができる。
　本発明のコク味付与剤、食品組成物、あるいは複合コク味付与剤は、食品、飲料、調味料等の各種飲食品に配合して用いることができる。
　本発明のコク味付与剤、食品組成物、あるいは複合コク味付与剤を食品、飲料、調味料等の各種飲食品に配合して用いる場合の最終的なγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙの量及び併用されるアミノ酸又はペプチドの量は所望の効果が得られる量であれば特に制限されないが、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙの量及び／又はアミノ酸若しくはペプチドの量として、食品、飲料あるいは調味料等の全質量を基準として、それぞれについて０．１ｐｐｂ～９９．９重量％、好ましくは１ｐｐｂ～１０質量％、より好ましくは０．０１ｐｐｍ～１質量％程度である。 The rich taste imparting agent, food composition, or complex rich taste imparting agent of the present invention can be used in any form without limitation on physical properties such as dry powder, paste, and solution.
The rich taste imparting agent, food composition, or complex rich taste imparting agent of the present invention can be used by blending it with various foods and beverages such as foods, beverages and seasonings.
The final amount of γ-Glu-Nva-Gly when the kokumi imparting agent, food composition, or complex kokumi imparting agent of the present invention is used in various foods and beverages such as foods, beverages and seasonings, and The amount of the amino acid or peptide used in combination is not particularly limited as long as the desired effect is obtained, but the amount of γ-Glu-Nva-Gly and / or the amount of amino acid or peptide may be food, beverage or seasoning, etc. From about 0.1 ppb to 99.9% by weight, preferably from 1 ppb to 10% by weight, and more preferably from about 0.01 ppm to 1% by weight.
　本発明のコク味付与剤、食品組成物、あるいは複合コク味付与剤が配合された食品、飲料、調味料等の各種飲食品には、飲食品的に許容しうるあらゆる固体又は液体の担体、適当な調味原料等をさらに配合させてもよい。
　上記担体としては、例えば、グルコース、乳糖、ショ糖、澱粉、マンニトール、デキストリン、脂肪酸グリセリド、ポリエチレングリコール、ヒドロキシエチルデンプン、エチレングリコール、ポリオキシエチレンソルビタン脂肪酸エステル、ゼラチン、アルブミン、アミノ酸、水、生理食塩水等が挙げられる。
　上記の調味原料は、当業界で用いられるいずれの調味原料であってもよく特に制限されないが、より具体的には既に上述のものが挙げられる。
　上記の担体、他の調味原料等はいずれもその含有量は特に制限されない。
　上記調味原料のうち、酵母エキスは、由来となる菌体・その培養条件・抽出処理方法のいずれも特に限定されず任意の酵母エキスを用いることができ、更に加熱処理、酵素処理、濃縮、粉末化処理等が施されたものでも良い。
　本発明は又、各種飲食品の製造中間品に、１質量ｐｐｂ～９９．９質量％含有させるようにγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを添加することを特徴とする、各種飲食品の製造方法を提供する。ここで各種飲食品としては、低脂肪食品が好ましい。 Various foods and beverages such as foods, beverages, and seasonings containing the rich body taste imparting agent, food composition, or complex body taste imparting agent of the present invention, any solid or liquid carrier that is acceptable for foods and beverages, An appropriate seasoning material or the like may be further blended.
Examples of the carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, gelatin, albumin, amino acid, water, and physiological saline. Water etc. are mentioned.
The seasoning raw material may be any seasoning raw material used in the art and is not particularly limited, but more specifically, the above-mentioned ones are already mentioned.
The content of any of the above carriers and other seasoning ingredients is not particularly limited.
Among the above-mentioned seasoning raw materials, the yeast extract is not particularly limited in any of the cells from which it is derived, its culture conditions, and the extraction treatment method, and any yeast extract can be used. Further, heat treatment, enzyme treatment, concentration, powder It may be one that has been processed.
The present invention also provides a method for producing various foods and drinks, characterized in that γ-Glu-Nva-Gly is added to the production intermediate product of various foods and drinks so that 1 mass ppb to 99.9% by mass is contained. provide. Here, as various foods and drinks, low-fat foods are preferable.
　本発明は又、各種飲食品の製造中間品に、本発明の食品組成物を添加することを特徴とする、各種飲食品の製造方法を提供する。ここで各種飲食品としては、低脂肪食品が好ましい。
　本発明のコク味付与剤を用いる製造中間品の製造方法については、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料（例えば、うま味原料、たん白加水分解物、畜肉エキス）に添加混合する工程、および、必要に応じて、得られる飲食品原料混合物をさらに調理する工程を含む、飲食品又は飲食品の製造中間品の製造方法が好ましい。
　ここで、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料に添加混合する工程が、飲食品の製造中間品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～９９９９００重量ｐｐｍ、好ましくは、０．１～２０００００重量ｐｐｍとする工程を含むのが好ましい。
　又、飲食品の製造中間品を別の飲食品原料（例えば、農産物、水産物、畜肉、乳製品、又は、それらの加工食品）に添加して、得られる飲食品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～５０重量ｐｐｍ、好ましくは、０．０５～２０重量ｐｐｍとする工程をさらに含むのが好ましい。
　又、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料に添加混合する工程が、飲食品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～５０重量ｐｐｍ、好ましくは、０．０５～２０重量ｐｐｍとする工程を含むのが好ましい。 This invention also provides the manufacturing method of various food-drinks characterized by adding the food composition of this invention to the manufacture intermediate goods of various food-drinks. Here, as various foods and drinks, low-fat foods are preferable.
Regarding the method for producing a production intermediate product using the body taste imparting agent of the present invention, a taste enhancer comprising γ-Glu-Nva-Gly is used as a raw material for foods and beverages (for example, umami raw material, protein hydrolyzate, animal meat extract). A method for producing food or drink or a food intermediate for producing food or drink, which includes a step of adding to and mixing, and, if necessary, a step of further cooking the resulting food or drink raw material mixture, is preferred.
Here, the step of adding and mixing the taste enhancer composed of γ-Glu-Nva-Gly to the raw material for the food and drink, the concentration of γ-Glu-Nva-Gly in the intermediate product of the food and drink is 0.01-999900 ppm by weight Preferably, it includes a step of adjusting to 0.1 to 200,000 ppm by weight.
In addition, the intermediate product of the food and drink is added to another food and drink raw material (for example, agricultural products, marine products, livestock meat, dairy products, or processed foods thereof), and the resulting food and drink γ-Glu-Nva-Gly It is preferable to further include a step of adjusting the concentration to 0.01 to 50 ppm by weight, preferably 0.05 to 20 ppm by weight.
Further, the step of adding and mixing the taste enhancer comprising γ-Glu-Nva-Gly to the food / beverage product raw material has a concentration of γ-Glu-Nva-Gly of the food / beverage product of 0.01 to 50 ppm by weight, preferably 0 It is preferable to include a step of setting the content to 0.05 to 20 ppm by weight.
　上記の製造方法において、飲食品がポーク原料を含有する食品であるのが好ましい。この場合、０．０１～５０重量ｐｐｍのγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙと、０．００５～８０重量％のポーク原料と、他の食品原料とを含有するのが好ましい。
　上記の製造方法において、飲食品がビーフ原料を含有する食品であるのが好ましい。この場合、０．０１～５０重量ｐｐｍのγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙと、０．００５～８０重量％のビーフ原料と、他の食品原料とを含有するのが好ましい。
　又、本発明の対象食品としては、上記の食品に加えて、アイスクリーム、蜂蜜、マーマレード、およびイチゴジャムなどの甘味が主のデザート・菓子など食品（スィート系食品）や、チキンスープなどの塩味が主の加工食品・惣菜・スナックなど食品（セイボリー系食品）も好ましいものとしてあげられる。
　以下に、実施例を挙げて本発明をさらに詳しく説明するが、これらは本発明を限定するものではない。 In said manufacturing method, it is preferable that food / beverage products are the foodstuffs containing a pork raw material. In this case, it is preferable to contain 0.01 to 50 ppm by weight of γ-Glu-Nva-Gly, 0.005 to 80% by weight of pork ingredients, and other food ingredients.
In said manufacturing method, it is preferable that food / beverage products are the foodstuffs containing a beef raw material. In this case, it is preferable to contain 0.01 to 50 ppm by weight of γ-Glu-Nva-Gly, 0.005 to 80% by weight of beef ingredients, and other food ingredients.
In addition to the above-mentioned foods, the target foods of the present invention are foods such as ice cream, honey, marmalade, and strawberry jam (sweet foods) such as desserts and confectionery (sweet foods), and salty tastes such as chicken soup However, foods such as processed foods, side dishes and snacks (savory foods) are also preferred.
Hereinafter, the present invention will be described in more detail with reference to examples, but these do not limit the present invention.
（合成例１）γ-Glu-Nva-Gly（γ-L-グルタミル-L-ノルバリル-グリシン）の合成
　Boc-Nva（t-Butoxycarbonyl-L-norvaline、4.44 g、20.4 mmol）とGly-OBzl・HCl（Glycine benzyl ester hydrochloride、4.12 g、20.4 mmol）を塩化メチレン（CH2Cl2、100 ml）に溶解した。反応液を0 ℃に保ち、トリエチルアミン（Et3N、3.13 ml、1.1当量、22.4 mmol）、HOBt・H2O（1-Hydroxybenzotriazole hydrate、3.44 g、1.1当量、22.4 mmol）及びWSC・HCl（1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride、4.30 g、1.1当量、22.4 mmol）を加えた。反応液の温度を徐々に昇温し、室温で一夜（16時間）攪拌した。反応液を減圧濃縮し、残渣に酢酸エチル（150 ml）を加え、有機層を、水（50 ml）、5 %クエン酸水溶液（50 ml）で2回、飽和食塩水（50 ml）、5 %炭酸水素ナトリウム水溶液（50 ml）で2回及び飽和食塩水（50 ml）で洗浄し、無水硫酸マグネシウムで乾燥した。硫酸マグネシウムをろ過して除き、ろ液を減圧濃縮した。残渣にn-ヘキサンを加えると結晶化したので、結晶をろ過して集め、減圧乾燥してBoc-Nva-Gly-OBzl (6.88 g、18.9 mmol)を結晶として得た。
　Boc-Nva-Gly-OBzl (6.88 g、18.9 mmol)に4N HCl / ジオキサン溶液（94.5 ml）を加え、室温で1時間攪拌した。減圧濃縮してジオキサンを除き、残渣にn-ヘキサン（30 ml）を加えて減圧濃縮する操作を3回繰り返して、定量的な収率でH-Nva-Gly-OBzlHClを得た。 (Synthesis Example 1) Synthesis of γ-Glu-Nva-Gly (γ-L-glutamyl-L-norvalyl-glycine) Boc-Nva (t-Butoxycarbonyl-L-norvaline, 4.44 g, 20.4 mmol) and Gly-OBzl HCl (Glycine benzyl ester hydrochloride, 4.12 g, 20.4 mmol) was dissolved in methylene chloride (CH 2 Cl 2 , 100 ml). The reaction solution was kept at 0 ° C., and triethylamine (Et 3 N, 3.13 ml, 1.1 eq, 22.4 mmol), HOBt · H 2 O (1-Hydroxybenzotriazole hydrate, 3.44 g, 1.1 eq, 22.4 mmol) and WSC · HCl (1 -Ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride, 4.30 g, 1.1 eq, 22.4 mmol) was added. The temperature of the reaction solution was gradually raised and stirred at room temperature overnight (16 hours). The reaction mixture was concentrated under reduced pressure, ethyl acetate (150 ml) was added to the residue, and the organic layer was washed twice with water (50 ml) and 5% aqueous citric acid solution (50 ml), saturated brine (50 ml), 5 The extract was washed twice with an aqueous sodium hydrogen carbonate solution (50 ml) and with saturated brine (50 ml), and dried over anhydrous magnesium sulfate. Magnesium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. Since the residue was crystallized by adding n-hexane, the crystals were collected by filtration and dried under reduced pressure to obtain Boc-Nva-Gly-OBzl (6.88 g, 18.9 mmol) as crystals.
To Boc-Nva-Gly-OBzl (6.88 g, 18.9 mmol) was added 4N HCl / dioxane solution (94.5 ml), and the mixture was stirred at room temperature for 1 hour. Dioxane was removed by concentration under reduced pressure, and the operation of adding n-hexane (30 ml) to the residue and concentrating under reduced pressure was repeated three times to obtain H-Nva-Gly-OBzlHCl in a quantitative yield.
　H-Nva-Gly-OBzlHClを塩化メチレン（130 ml）に溶解し、反応液を０ ℃に保った。反応液にZ-Glu-OBzl (N-α-Carbobenzoxy-L-glutamic acid α-benzyl ester、7.03 g、 18.9 mmol)、トリエチルアミン（2.90 ml、1.1当量、20.8 mmol）、HOBt・H2O（3.20 g、1.1当量、20.8 mmol）及びWSC・HCl（3.98 g、1.1当量、20.8 mmol）を加えた。反応液の温度を徐々に昇温し、室温で一夜（16時間）攪拌した。反応混合物を減圧濃縮し、残渣に酢酸エチル（1000 ml）を加え、有機層を、水（100 ml）、5 %クエン酸水溶液（100 ml）で2回、飽和食塩水（100 ml）、5 %炭酸水素ナトリウム水溶液（100 ml）で2回及び飽和食塩水（100 ml）で洗浄し、無水硫酸マグネシウムで乾燥した。溶液を50 ℃に加熱した後に、硫酸マグネシウムをろ過して除き、ろ液を減圧濃縮した。結晶が出始めたところでn-ヘキサンを加えて十分結晶を析出させた。結晶をろ過して集め、減圧乾燥してZ-Glu(Nva-Gly-OBzl)-OBzl (10.16 g、16.4 mmol)を結晶として得た。
　エタノール（250 ml）と水（30 ml）の混合液にZ-Glu(Nva-Gly-OBzl)-OBzl (10.16 g、16.4 mmol)と5 %パラジウム炭素（5 % palladium/carbon、1.20 g）を加え、50 ℃で一夜（14時間）、水素雰囲気下で接触還元を行った。反応中、水（100 ml）を少しずつ加えた。パラジウム炭素をろ過して除き、ろ液を減圧濃縮した。残渣を少量の水とエタノールから再結晶してγ-Glu-Nva-Gly (4.59 g、15.1 mmol)を白色結晶として得た。その特性値を次に示す。
ESI-MS:(M+H)+ = 304.1
1H-NMR (400 MHz, D2O) δ (ppm): 0.82 (3H, t, J=7.4 Hz), 1.23-1.37 (2H, m), 1.55-1.75 (2H, m), 2.01-2.09 (2H, m), 2.38-2.48 (2H, m), 3.72 (1H, t, J=6.4Hz), 3.87 (1H, dd, J=17.8  and 20.9Hz), 4.21 (1H, dd, J=4.4 and 8.9 Hz) H-Nva-Gly-OBzlHCl was dissolved in methylene chloride (130 ml), and the reaction solution was kept at 0 ° C. Z-Glu-OBzl (N-α-Carbobenzoxy-L-glutamic acid α-benzyl ester, 7.03 g, 18.9 mmol), triethylamine (2.90 ml, 1.1 eq, 20.8 mmol), HOBt · H 2 O (3.20 g, 1.1 eq, 20.8 mmol) and WSC.HCl (3.98 g, 1.1 eq, 20.8 mmol) were added. The temperature of the reaction solution was gradually raised and stirred at room temperature overnight (16 hours). The reaction mixture was concentrated under reduced pressure, ethyl acetate (1000 ml) was added to the residue, and the organic layer was washed twice with water (100 ml) and 5% aqueous citric acid solution (100 ml), saturated brine (100 ml), 5 The extract was washed twice with an aqueous sodium hydrogen carbonate solution (100 ml) and with saturated brine (100 ml), and dried over anhydrous magnesium sulfate. After the solution was heated to 50 ° C., magnesium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure. When crystals began to appear, n-hexane was added to sufficiently precipitate the crystals. The crystals were collected by filtration and dried under reduced pressure to give Z-Glu (Nva-Gly-OBzl) -OBzl (10.16 g, 16.4 mmol) as crystals.
Mix ethanol (250 ml) and water (30 ml) with Z-Glu (Nva-Gly-OBzl) -OBzl (10.16 g, 16.4 mmol) and 5% palladium on carbon (5% palladium / carbon, 1.20 g). In addition, catalytic reduction was carried out at 50 ° C. overnight (14 hours) under a hydrogen atmosphere. During the reaction, water (100 ml) was added little by little. Palladium carbon was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was recrystallized from a small amount of water and ethanol to obtain γ-Glu-Nva-Gly (4.59 g, 15.1 mmol) as white crystals. The characteristic values are as follows.
ESI-MS: (M + H) + = 304.1
1 H-NMR (400 MHz, D 2 O) δ (ppm): 0.82 (3H, t, J = 7.4 Hz), 1.23-1.37 (2H, m), 1.55-1.75 (2H, m), 2.01-2.09 (2H, m), 2.38-2.48 (2H, m), 3.72 (1H, t, J = 6.4Hz), 3.87 (1H, dd, J = 17.8 and 20.9Hz), 4.21 (1H, dd, J = 4.4 and 8.9 Hz)
（試験例１）ＣａＳＲ発現プラスミドの調製
　ＣａＳＲ発現プラスミドの調製を以下のように行った。
　ＮＣＢＩに登録されたＤＮＡ配列（ＣａＳＲ（カルシウム受容体）：NM_000388、配列番号１、２）を元に、ＰＣＲに使う合成オリゴＤＮＡ（フォワードプライマー（配列番号３：ACTAATACGACTCACTATAGGGACCATGGCATTTTATAGCTGCTGCTGG）、及びリバースプライマー（配列番号４:TTATGAATTCACTACGTTTTCTGTAACAG）を合成した。
　ヒト腎臓由来のｃＤＮＡ（Clontech社製）を材料として、前記プライマー、及びPfu Ultra DNA Polymerase（Stratagene社製）を用い、以下の条件でＰＣＲを実施した。９４℃で３分の後、９４℃で３０秒、５５℃で３０秒、７２℃で２分を３５回繰り返した後、７２℃で７分反応させた。アガロース電気泳動を行い、ＤＮＡ染色試薬で染色した後、紫外線照射によってＰＣＲによって増幅がなされたか否かを検出した。又、同時に電気泳動したサイズ既知のＤＮＡマーカーと比較することで、ＰＣＲ産物の鎖長を確認した。
　プラスミドベクターｐＢＲ３２２を制限酵素ＥｃｏＲＶ（Takara社製）によって切断し、その切断部位にＰＣＲによって増幅された遺伝子断片をLigation kit（Promega社製）を用いて連結した。この反応溶液でエシェリヒア・コリＤＨ５α株を形質転換し、ＰＣＲ増幅産物がクローニングされたプラスミドを保持する形質転換体を選抜し、更にＰＣＲ増幅産物をＤＮＡ塩基配列解析によって確認した。
　この組換えプラスミドを用いてヒトＣａＳＲ発現プラスミドhCaSR/pcDNA3.1を作製した。 (Test Example 1) Preparation of CaSR expression plasmid A CaSR expression plasmid was prepared as follows.
Based on the DNA sequence (CaSR (calcium receptor): NM_000388, SEQ ID NO: 1, 2) registered in NCBI, synthetic oligo DNA (forward primer (SEQ ID NO: 3: ACTAATACGACTCACTATAGGGACCATGGCATTTTATAGCTGCTGCTGG)) and reverse primer (SEQ ID NO: SEQ ID NO: 1) 4: TTATGAATTCACTACGTTTTCTGTAACAG) was synthesized.
PCR was carried out under the following conditions using cDNA derived from human kidney (manufactured by Clontech) as a material and the above primers and Pfu Ultra DNA Polymerase (manufactured by Stratagene). After 3 minutes at 94 ° C., 35 times of 94 ° C. for 30 seconds, 55 ° C. for 30 seconds and 72 ° C. for 2 minutes were repeated 35 times, followed by reaction at 72 ° C. for 7 minutes. After agarose electrophoresis and staining with a DNA staining reagent, it was detected whether or not amplification was performed by UV irradiation. In addition, the chain length of the PCR product was confirmed by comparison with a DNA marker of known size that was electrophoresed simultaneously.
Plasmid vector pBR322 was cleaved with restriction enzyme EcoRV (Takara), and the gene fragment amplified by PCR was ligated to the cleavage site using Ligation kit (Promega). The Escherichia coli DH5α strain was transformed with this reaction solution, and a transformant carrying a plasmid in which the PCR amplification product was cloned was selected, and the PCR amplification product was further confirmed by DNA nucleotide sequence analysis.
Using this recombinant plasmid, a human CaSR expression plasmid hCaSR / pcDNA3.1 was prepared.
（試験例２）CaSRアゴニスト活性の評価
　293E細胞（EBNA1発現HEK293細胞、ATCC No.CRL-10852）を、200μg/mlのG418（ジェネティシン）存在下、10%のウシ胎児血清を含むDMEM/Ham's-F12（3.15/ml Glucose含有Dulbecco's modified Eagle medium、ナカライテスク）にて培養した。3×106ceells/10mlでF25フラスコに撒き、CO2インキュベータ（5%CO2、37℃）に24時間静置した後、トランスフェクション試薬Fugene6（Roche）にてヒトCaSR発現プラスミドhCaSR/pcDNA3.1をトランスフェクションした。CO2インキュベータに6～7時間置いた後、細胞を10%ウシ胎児血清含有DMEM/Ham's-F12にて回収し、70,000cells/wellでpoly-D-lysine coat 96well plate（BD-Biocoat）に播種した。
　CO2インキュベータにて24時間静置した後、この細胞を播種した96 well plateから培地を除去し、Assay Buffer (146mM NaCl、5mM KCl、1mM MgSO4、1mg/ml Glucose、20mM HEPES(pH 7.2)、0.75～1.25 mM CaCl2)に溶解したCa2+蛍光指示薬Calcium 4 Assay Kit（Molecular Devices）を200μl/well添加し、37℃で1時間、次いで室温で10分静置し指示薬を取り込ませた。
　この96well plateに、0.1%BSA含有Assay Bufferに溶解した被験化合物を50μl/well添加し、FLEX Station（Molecular Devices）で3分間蛍光強度変化を測定した。 (Test Example 2) Evaluation of CaSR agonist activity 293E cells (EBNA1-expressing HEK293 cells, ATCC No. CRL-10852) were treated with DMEM / Ham's- containing 10% fetal bovine serum in the presence of 200 μg / ml G418 (Geneticin). The cells were cultured in F12 (Dulbecco's modified Eagle medium containing 3.15 / ml Glucose, Nacalai Tesque). Place in 3 x 10 6 ceells / 10 ml in a F25 flask, leave it in a CO 2 incubator (5% CO 2 , 37 ° C) for 24 hours, and then use human gene for transfection with the transfection reagent Fugene6 (Roche) hCaSR / pcDNA3. 1 was transfected. After 6-7 hours in a CO 2 incubator, cells were collected with DMEM / Ham's-F12 containing 10% fetal bovine serum and seeded on poly-D-lysine coat 96 well plate (BD-Biocoat) at 70,000 cells / well did.
After standing for 24 hours in a CO 2 incubator, remove the medium from the 96-well plate seeded with these cells, Assay Buffer (146 mM NaCl, 5 mM KCl, 1 mM MgSO 4 , 1 mg / ml Glucose, 20 mM HEPES (pH 7.2) 200 ul / well of Ca 2+ fluorescent indicator Calcium 4 Assay Kit (Molecular Devices) dissolved in 0.75 to 1.25 mM CaCl 2 ) was added and allowed to stand at 37 ° C. for 1 hour and then at room temperature for 10 minutes to incorporate the indicator. .
A test compound dissolved in 0.1% BSA-containing assay buffer was added to the 96-well plate at 50 μl / well, and the change in fluorescence intensity was measured with FLEX Station (Molecular Devices) for 3 minutes.
（EC50算出法）
　化合物添加前後の蛍光強度の最大値と最小値の差(RFU(Max-Min)）をFLEX Stationの自動計算にて求めた。化合物最大濃度添加時のRFU(Max-Min)を100%、被験化合物を含まない0.1%BSA含有Assay Bufferを使用時のRFU(Max-Min)を0%と定義した活性率を計算し、表計算ソフトXfitもしくはグラフパッドプリズムにてカーブフィッティングし、活性率50%時の化合物濃度であるEC50値を求めた。結果を表１に示した。また、比較例として、同様に測定したトリペプチドのデータを表２、表３に示した。なお、表３に記載のデータは非特許文献３に掲載されている。
　　　　　　　　表１
Figure JPOXMLDOC01-appb-I000002
(EC 50 calculation method)
The difference (RFU (Max-Min)) between the maximum value and the minimum value of the fluorescence intensity before and after the compound addition was determined by automatic calculation of FLEX Station. Calculate the activity rate by defining the RFU (Max-Min) at the time of adding the maximum compound concentration as 100%, and the RFU (Max-Min) as 0% when using 0.1% BSA-containing assay buffer that does not contain the test compound. Curve fitting was performed with the calculation software Xfit or the graph pad prism, and the EC 50 value, which is the compound concentration when the activity rate was 50%, was determined. The results are shown in Table 1. As comparative examples, data of tripeptides measured in the same manner are shown in Tables 2 and 3. The data shown in Table 3 is published in Non-Patent Document 3.
Table 1
Figure JPOXMLDOC01-appb-I000002
　　　　　　　　表２
Figure JPOXMLDOC01-appb-I000003
Table 2
Figure JPOXMLDOC01-appb-I000003
　　　　　　　　　表３
Figure JPOXMLDOC01-appb-I000004
　この結果から、γ-Glu-Nva-Glyは、他のγ-Glu-Ｘ-Gly型の構造を有するペプチドと同程度に高いＣａＳＲ活性を有することを予期せず見出した。 Table 3
Figure JPOXMLDOC01-appb-I000004
From this result, it was unexpectedly found that γ-Glu-Nva-Gly has CaSR activity as high as that of other peptides having a γ-Glu-X-Gly type structure.
実施例１　コク味付与活性の評価
　γ-Glu-Nva-Glyについて、定量的な官能評価試験によりコク味付与活性の強度を調べた。
　定量的官能評価試験は以下のように実施した。グルタミン酸ナトリウム（０．０５ｇ／ｄｌ）、イノシン酸一リン酸（０．０５ｇ／ｄｌ）、塩化ナトリウム（０．５ｇ／ｄｌ）を含有する蒸留水に、試料として化合物類を０．０００００１～０．１ｇ／ｄｌにて混合した場合の、コク味付与活性の強度を測定した。試料溶解後に無添加コントロールに対し酸性を呈したサンプルについては、ＮａＯＨで無添加コントロールに対しｐＨ±０．２の幅に合わせて使用した。官能評点について、コントロール：０点、強い：３点、非常に強い：５点とするとともに、尺度をより明確にするため、γ-Glu-Cys-Glyの先中味、後味を各々3.0点とした。採点ついては、直線尺度法を用い、－５～０～５点の位置を示した直線に対し、該当する採点を位置として記入する方法を用いた。また、食品の調味開発を累積で１年以上経験し、うま味塩味溶液に添加したγ-Glu-Cys-Glyとγ-Glu-Val-Glyの力価の差が１０倍前後と判定できる者（定期的に確認）をパネラーとした。評価は、ｎ＝４で実施した。尚、「先中味」とは、***み後、0～5秒の呈味、後味はそれ以降の呈味である。被検化合物は、上記添加濃度で幅広くコク味付与活性を示したが、代表的な濃度の結果を表４に示した。
　この結果、γ-Glu-Nva-Gly以外のトリペプチドについては、いずれも最大でグルタチオン（γ-Glu-Cys-Gly）の約１０倍程度の活性であったが、驚くべきことにγ-Glu-Nva-Glyは、さらに高く、１００倍強の高活性を有することが示された。 Example 1 Evaluation of kokumi imparting activity The strength of kokumi imparting activity of γ-Glu-Nva-Gly was examined by a quantitative sensory evaluation test.
The quantitative sensory evaluation test was performed as follows. In a distilled water containing sodium glutamate (0.05 g / dl), inosinic acid monophosphate (0.05 g / dl), sodium chloride (0.5 g / dl), the compounds as a sample were 0.000001-0. The strength of the savoriness imparting activity when mixed at 1 g / dl was measured. About the sample which showed the acidity with respect to the additive-free control after sample dissolution, it was used according to the range of pH ± 0.2 with respect to the additive-free control with NaOH. For sensory scores, control: 0 points, strong: 3 points, very strong: 5 points, and γ-Glu-Cys-Gly's pre-taste and after-taste were set to 3.0 points to make the scale clearer. . Regarding the scoring, a linear scaling method was used, and a method of entering the corresponding scoring as a position on a straight line indicating positions of −5 to 0 to 5 points was used. Those who have experienced the development of seasoning for food for more than a year and can determine that the difference in titer between γ-Glu-Cys-Gly and γ-Glu-Val-Gly added to the umami salty solution is around 10 times ( Regularly confirmed) was the panel. Evaluation was carried out with n = 4. The “first taste” refers to a taste of 0 to 5 seconds after the mouth, and the aftertaste is a taste after that. The test compounds exhibited a wide range of richness-imparting activity at the above addition concentrations. Table 4 shows the results of typical concentrations.
As a result, all the tripeptides other than γ-Glu-Nva-Gly were about 10 times as high as glutathione (γ-Glu-Cys-Gly), but surprisingly, γ-Glu -Nva-Gly was even higher and was shown to be 100 times more highly active.
　　　　　　　　　　　　　　　　表４
Figure JPOXMLDOC01-appb-I000005
Table 4
Figure JPOXMLDOC01-appb-I000005
　γ－Glu－Nva-Glyはγ-Glu-Cys-Glyに比較して約100倍、γ-Glu-Val-Glyなどに比べて、コク味付与活性が少なくとも10倍程度高い。更に、γ-Glu-Ala-Gly、γ-Glu-Abu-Gly、γ-Glu-ｔLeu-Glyのように、異味（後味の収斂味など）なく、γ-Glu-Nva-Glyが優れていることが分かった。 Γ-Glu-Nva-Gly is about 100 times higher than γ-Glu-Cys-Gly, and at least 10 times higher in taste-providing activity than γ-Glu-Val-Gly. Furthermore, γ-Glu-Nva-Gly is superior, with no off-flavors (such as aftertaste astringency), such as γ-Glu-Ala-Gly, γ-Glu-Abu-Gly, and γ-Glu-tLeu-Gly. I understood that.
実施例２　コク味付与活性の評価
　γ-Glu-Nva-Glyについて、中後味型を明確にするため別評価項目での定量的な官能評価試験によりコク味付与活性の強度を調べた。
　定量的官能評価試験は以下のように実施した。中後味を分かり易くするため、評価液に関し、イノシン酸一リン酸を使用せず、中後のうま味を低減した。すなわち、グルタミン酸ナトリウム（０．１ｇ／ｄｌ）、塩化ナトリウム（０．４ｇ／ｄｌ）を含有する蒸留水に、試料として化合物類を０．０００００１～０．１ｇ／ｄｌにて混合した場合の、コク味付与活性の強度を測定した。試料溶解後に無添加コントロールに対し酸性を呈したサンプルについては、ＮａＯＨで無添加コントロールに対しｐＨ±０．２の幅に合わせて使用した。官能評点について、コントロール：０点、強い：３点、非常に強い：５点とするとともに、尺度をより明確にするため、γ-Glu-Cys-Glyの先、中後味を各々3.0点とした。採点ついては、直線尺度法を用い、－５～０～５点を示した直線に対し、該当する採点を位置として示す方法を用いた。また、食品の調味開発を累積で１年以上経験し、うま味塩味溶液に添加したγ-Glu-Cys-Glyとγ-Glu-Val-Glyの力価の差が１０倍前後と判定できる者（定期的に確認）をパネラーとした。評価は、ｎ＝４で実施した。尚、「先味」とは、***み後、0～2秒の呈味、中後味はそれ以降の呈味である。被検化合物は、上記添加濃度で幅広くコク味付与活性を示したが、代表的な濃度の結果を表５に示した。
　この結果でも、γ-Glu-Val-Glyは、グルタチオン（γ-Glu-Cys-Gly）の約１０倍程度の活性であったが、γ-Glu-Nva-Glyは、さらに高く、１００倍強の高活性を有することが示された。 Example 2 Evaluation of kokumi imparting activity For γ-Glu-Nva-Gly, the strength of kokumi imparting activity was examined by a quantitative sensory evaluation test in another evaluation item in order to clarify the middle aftertaste type.
The quantitative sensory evaluation test was performed as follows. In order to make the middle aftertaste easier to understand, the innocous acid monophosphate was not used in the evaluation solution, and the umami after the middle was reduced. That is, when the compounds as a sample were mixed at 0.000001 to 0.1 g / dl in distilled water containing sodium glutamate (0.1 g / dl) and sodium chloride (0.4 g / dl), The intensity of taste imparting activity was measured. About the sample which showed the acidity with respect to the additive-free control after sample dissolution, it was used according to the range of pH ± 0.2 with respect to the additive-free control with NaOH. For sensory scores, control: 0 points, strong: 3 points, very strong: 5 points, and γ-Glu-Cys-Gly tip and middle aftertaste were set to 3.0 points respectively to make the scale clearer. . Regarding the scoring, a linear scaling method was used, and a method of showing the corresponding scoring as a position with respect to a straight line indicating −5 to 0 to 5 points was used. Those who have experienced the development of seasoning for food for more than a year and can determine that the difference in titer between γ-Glu-Cys-Gly and γ-Glu-Val-Gly added to the umami salty solution is around 10 times ( Regularly confirmed) was the panel. Evaluation was carried out with n = 4. The “prior taste” is a taste of 0 to 2 seconds after the mouth is contained, and the medium after taste is a taste after that. The test compounds exhibited a wide range of richness-imparting activity at the above-mentioned added concentrations. Table 5 shows the results of typical concentrations.
Even in this result, γ-Glu-Val-Gly was about 10 times as active as glutathione (γ-Glu-Cys-Gly), but γ-Glu-Nva-Gly was even higher, slightly more than 100 times. It was shown to have a high activity.
　　　　　　　　　　　　　　　　表５
Figure JPOXMLDOC01-appb-I000006
Table 5
Figure JPOXMLDOC01-appb-I000006
　γ-Glu-Nva-Glyが優れたコク味付与活性を有し、その呈味パターンについて中後味の特質を有し、異味（収斂味など）なく優れていることが分かった。また、γ-Glu-Nva-Glyはγ-Glu-Cys-Glyに比較して約100倍、γ-Glu-Val-Glyなどに比べて、コク味付与活性が少なくとも10倍程度高いため、きわめて低濃度での使用が可能である。したがって、より簡便かつ低コストでコク味付与剤を提供することを可能にし、産業上も非常に有利である。 It was found that γ-Glu-Nva-Gly has an excellent kokumi imparting activity, has a taste of medium aftertaste, and is excellent with no off-flavors (such as astringent taste). In addition, γ-Glu-Nva-Gly is about 100 times higher than γ-Glu-Cys-Gly, and has a kokumi imparting activity that is at least 10 times higher than γ-Glu-Val-Gly. It can be used at low concentrations. Therefore, it is possible to provide a richness-imparting agent more easily and at a low cost, which is very advantageous from an industrial viewpoint.
実施例３　食品におけるコク味付与活性の評価
　γ-Glu-Nva-Glyについて、実際、食品に用いた場合、高力価なγ-Glu-Val-Glyより、効果が極めて高いかどうかを官能評価試験により調べた。
　官能評価試験は以下のように実施した。中後味が強いと考えられる食品として、甘味が主のデザート・菓子など食品（スィート系食品）の代表として、市販されているアイスクリーム、蜂蜜、マーマレード、およびイチゴジャムを使用した。塩味が主の加工食品・惣菜・スナックなど食品（セイボリー系食品）の代表として市販のチキンスープ、０．１重量％市販粉末コショーをマッシュポテトに加えたペースト、市販ねりショウガ、および２重量％バターをマッシュポテトに加えたペーストを使用した。比較するγ-Glu-Val-Glyの混合量について、効果が明白な０．００２重量％とした。γ-Glu-Nva-Glyについて、０．００００００１～０．０１重量％にて混合した場合の、呈味全体の増強（コク味付与活性の強度）を測定した。コントロールは無添加の食品である。官能評点について、小数点の差など小差を生まないようコントロール：±、やや強い：＋、強い：＋＋,非常に強い＋＋＋とした。＋を１、＋＋を２、＋＋＋を３とし、数値平均が２．２ならば２＝＋＋など、四捨五入した。また、食品の調味開発を累積で１年以上経験し、うま味塩味溶液に添加したγ-Glu-Cys-Glyとγ-Glu-Val-Glyの力価の差が１０倍前後と判定できる者（定期的に確認）をパネラーとした。評価は、ｎ＝４で実施した。γ-Glu-Val-Glyについて、上記添加濃度で幅広くコク味付与活性を示したが、明確に比較できる濃度の結果を表７と表８に示した。
　この結果からも、グルタチオン（γ-Glu-Cys-Gly）の約１０倍程度のコク味付与活性であるγ-Glu-Val-Glyに対し、γ-Glu-Nva-Glyは、さらに５～１３倍強と著しく高い活性を有することが示された。 Example 3 Evaluation of kokumi- taste-imparting activity in foods γ-Glu-Nva-Gly is actually sensory-evaluated whether it is extremely effective compared to high-titer γ-Glu-Val-Gly when used in foods. It was examined by testing.
The sensory evaluation test was performed as follows. Commercially available ice cream, honey, marmalade, and strawberry jam were used as representative foods such as desserts and confectionery that mainly have sweetness (sweet foods) as foods that are thought to have a strong aftertaste. As a representative of processed foods such as salty foods, side dishes, and snacks (savory foods), a commercially available chicken soup, 0.1% by weight of powdered powdered pepper and mashed potato paste, commercially available ginger, and 2% by weight of butter The paste added to the mashed potato was used. The amount of γ-Glu-Val-Gly to be compared was set to 0.002% by weight where the effect was obvious. When γ-Glu-Nva-Gly was mixed at 0.0000001 to 0.01% by weight, the enhancement of the overall taste (strength of kokumi imparting activity) was measured. The control is an additive-free food. For sensory scores, control: ±, slightly strong: +, strong: ++, very strong ++, so as not to produce small differences such as decimal point differences. + Was set to 1, ++ was set to 2, ++ was set to 3, and if the numerical average was 2.2, 2 = ++ was rounded off. Those who have experienced the development of seasoning for food for more than a year and can determine that the difference in titer between γ-Glu-Cys-Gly and γ-Glu-Val-Gly added to the umami salty solution is around 10 times ( Regularly confirmed) was the panel. Evaluation was carried out with n = 4. With respect to γ-Glu-Val-Gly, the richness-enhancing activity was widely exhibited at the above-mentioned added concentrations. Tables 7 and 8 show the results of concentrations that can be clearly compared.
This result also shows that γ-Glu-Nva-Gly has an additional 5-13 compared to γ-Glu-Val-Gly, which is about 10 times as rich as glutathione (γ-Glu-Cys-Gly). It was shown to have a double strength and extremely high activity.
　　　　　　　　　　　　　　　　　　表７
Figure JPOXMLDOC01-appb-I000007
Table 7
Figure JPOXMLDOC01-appb-I000007
　　　　　　　　　　　　　　　　　　　表８
Figure JPOXMLDOC01-appb-I000008
Table 8
Figure JPOXMLDOC01-appb-I000008
　γ-Glu-Nva-Glyについて、実際のセイボリー系からスィート系食品までの中後味が特徴的なすべての食品において、全体の呈味を上げる有望なコク味付与活性を有することが分かった。さらにγ-Glu-Nva-Glyは、実際の食品において、γ-Glu-Val-Glyに比し、５～１３倍強と著しく高いコク味付与活性を有することが示された。したがって、改善を行いたいが、品質安定性のため、これ以上原料を配合できない場合でも、γ-Glu-Nva-Glyを用いることにより、極微量で品質改善が可能である。また、低コストでコク味付与剤を提供することが可能である。 Γ-Glu-Nva-Gly was found to have promising kokumi imparting activity that enhances the overall taste in all foods characterized by a medium aftertaste from actual savory to sweet foods. Furthermore, it was shown that γ-Glu-Nva-Gly has an extremely high body taste imparting activity of 5 to 13 times as much as γ-Glu-Val-Gly in actual foods. Therefore, it is desired to improve the quality. However, even if the raw materials cannot be blended any more because of the quality stability, the quality can be improved in a very small amount by using γ-Glu-Nva-Gly. Moreover, it is possible to provide a rich taste imparting agent at low cost.
実施例４　γ－Glu－Nva－Glyのポークエキスに与える効果
　γ－Glu－Nva－Glyについて、コク味付与効果活性がγ－Glu－Cys－Gly（グルタチオン）やγ－Glu－Val－Glyより喫食後、早い時間から強まることがわかった。よって、完全な中味型ではなくやや早く呈味が強まるポークエキスに対し、γ－Glu－Nva－Glyが、γ－Glu－Cys－Gly（グルタチオン）やγ－Glu－Val－Glyより著しく効果的であることを官能評価試験により調べた。
　官能評価試験は以下のように実施した。市販ポークエキス（固形分５５．１重量％、塩分９．３重量％）を５．０重量％となるように熱水に溶解し、ポークエキス溶液を調製した。このポークエキス溶液に対し、試料としてγ－Glu－Nva－Gly、γ－Glu－Cys－Gly、又はγ－Glu－Val－Glyを混合した。測定は２点識別試験法を用い、(1)γ－Glu－Nva－Gly０．０００３重量％と同等のコク味付与活性であるγ－Glu－Cys－Gly０．０２重量％、(2)γ－Glu－Nva－Gly０．０００３重量％と同等量であるγ－Glu－Val-Gly０．０００３重量％、(3)γ－Glu－Nva－Gly０．０００３重量％と同等のコク味付与活性であるγ－Glu－Val－Gly０．００２重量％、を比較評価し、“ポークエキスを呈味・風味のバランス変えず強め好ましい”方を判断させた。評価はN=９で実施した。γ－Glu－Nva－Gly０．０００３重量％の方が“ポークエキスを呈味・風味のバランスを変えず強め好ましい”と評価したパネル数を表９に示した。
　この結果から(1)と(3)のように、同等のコク味力価でも、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙのほうが明らかに“ポークエキスを呈味・風味のバランスを変えずに強め好ましい”ことが示された。 Example 4 Effect of γ-Glu-Nva-Gly on pork extract As for γ-Glu-Nva-Gly, kokumi imparting activity activity is higher than γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly After eating, it turned out that it strengthened from an early time. Therefore, γ-Glu-Nva-Gly is significantly more effective than γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly for pork extracts that are not completely medium-type and have a slightly stronger taste. It was examined by a sensory evaluation test.
The sensory evaluation test was performed as follows. A commercially available pork extract (solid content 55.1 wt%, salt content 9.3 wt%) was dissolved in hot water so as to be 5.0 wt% to prepare a pork extract solution. To this pork extract solution, γ-Glu-Nva-Gly, γ-Glu-Cys-Gly, or γ-Glu-Val-Gly was mixed as a sample. The measurement was performed using a two-point discrimination test method. (1) γ-Glu-Cys-Gly 0.02% by weight, which has a kokumi-imparting activity equivalent to γ-Glu-Nva-Gly 0.0003% by weight, (2) γ- Γ-Glu-Val-Gly 0.0003% by weight equivalent to Glu-Nva-Gly 0.0003% by weight, (3) γ having a savoring activity equivalent to γ-Glu-Nva-Gly 0.0003% by weight -Glu-Val-Gly 0.002% by weight was compared and evaluated to determine whether the "pork extract is stronger and preferable without changing the balance between taste and flavor". Evaluation was performed at N = 9. Table 9 shows the number of panels in which γ-Glu-Nva-Gly 0.0003% by weight was evaluated as “poke extract is stronger and more favorable without changing the balance between taste and flavor”.
From these results, it is clear that γ-Glu-Nva-Gly clearly shows that “Poke extract is strengthened and favored without changing the balance between taste and flavor” even at the equivalent strength value as in (1) and (3). It was shown that.
　　　　　　　　　　　　　　　　　表９
Ｎ＝９
Figure JPOXMLDOC01-appb-I000009
*)有意水準５％でγ-Glu-Nva-Glyの方が、ポークエキスを呈味・風味のバランスを変えずに強め好ましいと言える。 Table 9
N = 9
Figure JPOXMLDOC01-appb-I000009
*) It can be said that γ-Glu-Nva-Gly is stronger and more preferable at 5% significance level without changing the balance between taste and flavor of pork extract.
　γ－Glu－Nva－Glyについて、同等コク味付与活性である濃度のγ－Glu－Cys－Glyやγ－Glu－Val－Glyよりも“ポークエキスを呈味・風味のベースのバランスを変えずに強め好ましくすること”という極めて特異的な効果を有することが分った。ポーク原料は世界的に広く、調味料・スープ・畜肉加工品・調理加工品・菓子・スナックなどに広く用いられている。従って、γ－Glu－Nva－Glyはより低コストおよび微量で、食品を改善することを可能とし、産業上も非常に有利である。 Compared to γ-Glu-Cys-Gly and γ-Glu-Val-Gly at the same concentration level as γ-Glu-Nva-Gly, the pork extract does not change the balance between taste and flavor base. It has been found that it has a very specific effect of “strengthening and favoring”. Pork ingredients are widely used worldwide for seasonings, soups, processed meat products, cooked products, confectionery, snacks, etc. Therefore, γ-Glu-Nva-Gly can improve foods at a lower cost and in a minute amount, and is very advantageous from an industrial viewpoint.
実施例５　γ－Glu－Nva－Glyのビーフエキスに与える効果
　γ－Glu－Nva－Glyについて、コク味付与効果活性がγ－Glu－Cys－Gly（グルタチオン）やγ－Glu－Val－Glyより喫食後、早い時間から強まることがわかった。よって、完全な中味型ではなくやや早く呈味が強まり後味まで続くビーフエキスに対し、γ－Glu－Nva－Glyが、γ－Glu－Cys－Gly（グルタチオン）やγ－Glu－Val－Glyより著しく効果的であることを官能評価試験により調べた。
　官能評価試験は以下のように実施した。市販ビーフエキス（固形分６１．２重量％、塩分１２．２重量％）を３．０重量％となるように熱水に溶解し、ビーフエキス溶液を調製した。このビーフエキス溶液に対し、試料としてγ－Glu－Nva－Gly、γ－Glu－Cys－Gly、又はγ－Glu－Val－Glyを混合した。測定は２点識別試験法を用い、(1)γ－Glu－Nva－Gly０．０００３重量％と同等のコク味付与活性であるγ－Glu－Cys－Gly０．０２重量％、(2)γ－Glu－Nva－Gly０．０００３重量％と同等量であるγ－Glu－Val-Gly０．０００３重量％、(3)γ－Glu－Nva－Gly０．０００３重量％と同等のコク味付与活性であるγ－Glu－Val－Gly０．００２重量％、を比較評価し、“ビーフエキスを呈味・風味のバランス変えず強め好ましい”方を判断させた。評価はN=９で実施した。γ－Glu－Nva－Gly０．０００３重量％の方が“ビーフエキスを呈味・風味のバランスを変えず強め好ましい”と評価したパネル数を表１０に示した。
　この結果から(1)と(3)のように、同等のコク味力価でも、γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙのほうが明らかに“ビーフエキスを呈味・風味のバランスを変えずに強め好ましい”ことが示された。 Example 5 Effect of γ-Glu-Nva-Gly on Beef Extract With γ-Glu-Nva-Gly, Kokumi imparting activity is eaten from γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly Later, it turned out that it strengthened from an early time. Therefore, γ-Glu-Nva-Gly is significantly more γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly than beef extract, which is not a perfect middle-type but has a slightly faster taste and lasts until aftertaste. Effectiveness was examined by a sensory evaluation test.
The sensory evaluation test was performed as follows. A commercially available beef extract (solid content: 61.2 wt%, salt content: 12.2 wt%) was dissolved in hot water to 3.0 wt% to prepare a beef extract solution. To this beef extract solution, γ-Glu-Nva-Gly, γ-Glu-Cys-Gly, or γ-Glu-Val-Gly was mixed as a sample. The measurement was performed using a two-point discrimination test method. (1) γ-Glu-Cys-Gly 0.02% by weight, which has a kokumi-imparting activity equivalent to γ-Glu-Nva-Gly 0.0003% by weight, (2) γ- Γ-Glu-Val-Gly 0.0003% by weight equivalent to Glu-Nva-Gly 0.0003% by weight, (3) γ having a savoring activity equivalent to γ-Glu-Nva-Gly 0.0003% by weight -Glu-Val-Gly 0.002% by weight was comparatively evaluated to determine whether "beef extract is stronger and preferable without changing the balance between taste and flavor". Evaluation was performed at N = 9. Table 10 shows the number of panels in which γ-Glu-Nva-Gly 0.0003% by weight was evaluated as “a beef extract is stronger and preferable without changing the balance between taste and flavor”.
From these results, it is clear that γ-Glu-Nva-Gly clearly “strengthens and favors beef extract without changing the balance of taste and flavor” even with the same rich taste strength as in (1) and (3) It has been shown.
　　　　　　　　　　　　　　　　　表１０
　Ｎ＝９
Figure JPOXMLDOC01-appb-I000010
*)有意水準１％でγ-Glu-Nva-Glyの方が、ビーフエキスを呈味・風味のバランスを変えずに強め好ましいと言える。
**)有意水準５％でγ-Glu-Nva-Glyの方が、ビーフエキスを呈味・風味のバランスを変えずに強め好ましいと言える。 Table 10
N = 9
Figure JPOXMLDOC01-appb-I000010
*) It can be said that γ-Glu-Nva-Gly is stronger and preferable without changing the balance between taste and flavor at a significance level of 1%.
**) At a significance level of 5%, γ-Glu-Nva-Gly strengthens beef extract without changing the balance between taste and flavor, and is preferable.
　γ－Glu－Nva－Glyについて、同等コク味付与活性である濃度のγ－Glu－Cys－Glyやγ－Glu－Val－Glyよりも“ビーフエキスを呈味・風味のベースのバランスを変えずに強め好ましくすること”という極めて特異的な効果を有することが分った。ビーフ原料は世界的に広く、調味料・スープ・畜肉加工品・調理加工品・菓子・スナックなどに広く用いられている。従って、γ－Glu－Nva－Glyはより低コストおよび微量で、食品を改善することを可能とし、産業上も非常に有利である。 Compared to γ-Glu-Cys-Gly and γ-Glu-Val-Gly at the same concentration level as γ-Glu-Nva-Gly, “beef extract can be used without changing the balance between taste and flavor base. It has been found that it has a very specific effect of “enhancing and favoring”. Beef ingredients are widely used worldwide for seasonings, soups, processed meat products, cooked products, confectionery, snacks, etc. Therefore, γ-Glu-Nva-Gly can improve foods at a lower cost and in a minute amount, and is very advantageous from an industrial viewpoint.

Claims (14)

  1. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなるコク味付与剤。 A richness imparting agent comprising γ-Glu-Nva-Gly.
  2. 　（ａ）γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙに、
    　（ｂ）γ－Ｇｌｕ－Ｘ－Ｇｌｙ（Ｘはアミノ酸又はアミノ酸誘導体を表す）、γ－Ｇｌｕ－Ｖａｌ－Ｙ（Ｙはアミノ酸又はアミノ酸誘導体を表す）、γ－Ｇｌｕ－Ｎｖａ、γ－Ｇｌｕ－Ａｂｕ、γ－Ｇｌｕ－Ａｌａ、γ－Ｇｌｕ－Ｇｌｙ、γ－Ｇｌｕ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ、γ－Ｇｌｕ－Ｔｈｒ、γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｏｒｎ、Ａｓｐ－Ｇｌｙ、Ｃｙｓ－Ｇｌｙ、Ｃｙｓ－Ｍｅｔ、Ｇｌｕ－Ｃｙｓ、Ｇｌｙ－Ｃｙｓ、Ｌｅｕ－Ａｓｐ、Ｄ－Ｃｙｓ、γ－Ｇｌｕ－Ｍｅｔ（Ｏ）、γ－Ｇｌｕ－γ－Ｇｌｕ－Ｖａｌ、γ－Ｇｌｕ－Ｖａｌ－ＮＨ2、γ－Ｇｌｕ－Ｖａｌ－ｏｌ、γ－Ｇｌｕ－Ｓｅｒ、γ－Ｇｌｕ－Ｔａｕ、γ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）（Ｏ）、γ－Ｇｌｕ－Ｌｅｕ、γ－Ｇｌｕ－Ｉｌｅ、γ－Ｇｌｕ－ｔ－Ｌｅｕおよびγ－Ｇｌｕ－Ｃｙｓ（Ｓ－Ｍｅ）からなる群より選択される１種又は２種以上のアミノ酸又はペプチド、を併用してなる複合コク味付与剤。     (A) To γ-Glu-Nva-Gly,
    (B) γ-Glu-X-Gly (X represents an amino acid or amino acid derivative), γ-Glu-Val-Y (Y represents an amino acid or amino acid derivative), γ-Glu-Nva, γ-Glu-Abu , Γ-Glu-Ala, γ-Glu-Gly, γ-Glu-Cys, γ-Glu-Met, γ-Glu-Thr, γ-Glu-Val, γ-Glu-Orn, Asp-Gly, Cys-Gly Cys-Met, Glu-Cys, Gly-Cys, Leu-Asp, D-Cys, γ-Glu-Met (O), γ-Glu-γ-Glu-Val, γ-Glu-Val-NH 2 , γ -Glu-Val-ol, γ-Glu-Ser, γ-Glu-Tau, γ-Glu-Cys (S-Me) (O), γ-Glu-Leu, γ-Glu-Ile, γ-Glu-t -Leu and γ -A complex body taste imparting agent comprising one or more amino acids or peptides selected from the group consisting of Glu-Cys (S-Me).
  3. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを０．１ｐｐｂ～９９．９重量％含有する食品組成物。 A food composition containing 0.1 ppb to 99.9% by weight of γ-Glu-Nva-Gly.
  4. 　０．０１～５０重量ｐｐｍのγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙと、０．００５～８０重量％のポーク原料と、他の食品原料とを含有する、請求項３記載の食品組成物。 The food composition according to claim 3, comprising 0.01 to 50 ppm by weight of γ-Glu-Nva-Gly, 0.005 to 80% by weight of pork ingredients, and other food ingredients.
  5. 　０．０１～５０重量ｐｐｍのγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙと、０．００５～８０重量％のビーフ原料と、他の食品原料とを含有する、請求項３記載の食品組成物。 The food composition according to claim 3, comprising 0.01 to 50 ppm by weight of γ-Glu-Nva-Gly, 0.005 to 80% by weight of beef ingredients, and other food ingredients.
  6. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料に添加混合する工程、および、必要に応じて、得られる飲食品原料混合物をさらに調理する工程を含む、飲食品又は飲食品の製造中間品の製造方法。 A method for adding food and beverages or foods and beverages comprising a step of adding and mixing a taste enhancer comprising γ-Glu-Nva-Gly to food and beverage materials and, if necessary, further cooking the resulting food and beverage materials mixture Manufacturing method for intermediate products.
  7. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料に添加混合する工程が、飲食品の製造中間品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～９９９,９００重量ｐｐｍとする工程を含む、請求項６記載の飲食品又は飲食品原料の製造中間品の製造方法。 The step of adding and mixing a taste enhancer consisting of γ-Glu-Nva-Gly to the raw material for food and drink is such that the concentration of γ-Glu-Nva-Gly in the intermediate product of the food and drink is 0.01 to 999,900 ppm by weight. The manufacturing method of the manufacturing intermediate goods of the food-drinks or food-drinks raw material of Claim 6 including the process to do.
  8. 　飲食品の製造中間品を別の飲食品原料に添加して、得られる飲食品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～５０重量ｐｐｍとする工程をさらに含む、請求項７記載の飲食品の製造方法。 8. The method according to claim 7, further comprising the step of adding a food / beverage product manufacturing intermediate product to another food / beverage product raw material so that the resulting food / beverage product has a γ-Glu-Nva-Gly concentration of 0.01 to 50 ppm by weight. The manufacturing method of food-drinks.
  9. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙからなる呈味増強剤を飲食品原料に添加混合する工程が、飲食品のγ－Ｇｌｕ－Ｎｖａ－Ｇｌｙ濃度を０．０１～５０重量ｐｐｍとする工程を含む、請求項７記載の飲食品の製造方法。 The step of adding and mixing the taste enhancer composed of γ-Glu-Nva-Gly to the raw material for food and drink includes the step of setting the γ-Glu-Nva-Gly concentration of the food and drink to 0.01 to 50 ppm by weight. Item 8. A method for producing a food or drink according to Item 7.
  10. 　飲食品がポーク原料を含有する食品である、請求項６～９のいずれか１項に記載の飲食品の製造方法。 The method for producing a food or drink according to any one of claims 6 to 9, wherein the food or drink is a food containing a pork raw material.
  11. 　飲食品がビーフ原料を含有する食品である、請求項６～１０のいずれか１項に記載の飲食品の製造方法。 The method for producing a food or drink according to any one of claims 6 to 10, wherein the food or drink is a food containing beef ingredients.
  12. 　請求項６～１１のいずれかに記載の方法により得られる飲食品又は飲食品の製造中間品。 A food or drink obtained by the method according to any one of claims 6 to 11, or an intermediate product of the food or drink.
  13. 　γ－Ｇｌｕ－Ｎｖａ－Ｇｌｙを含有する組成物を飲食品に添加する工程を有する、飲食品の呈味増強方法。 A method for enhancing the taste of a food or drink, comprising a step of adding a composition containing γ-Glu-Nva-Gly to the food or drink.
  14. 　呈味増強がコク味付与である、請求項１３記載の方法。 The method according to claim 13, wherein the taste enhancement is richness imparting.
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WO2013118742A1 (en) 2012-02-06 2013-08-15 味の素株式会社 Composition for imparting body taste to foods and drinks
WO2013133404A1 (en) * 2012-03-09 2013-09-12 味の素株式会社 Seasoning
WO2014017485A1 (en) * 2012-07-25 2014-01-30 味の素株式会社 Food or beverage containing fruit juice
JP2016019470A (en) * 2014-07-11 2016-02-04 味の素株式会社 W/o/w type emulsion
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