CN102753041B - Dense taste imparting agent - Google Patents
Dense taste imparting agent Download PDFInfo
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- CN102753041B CN102753041B CN201080063730.6A CN201080063730A CN102753041B CN 102753041 B CN102753041 B CN 102753041B CN 201080063730 A CN201080063730 A CN 201080063730A CN 102753041 B CN102753041 B CN 102753041B
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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
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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
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- 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
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- 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
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- 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
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Abstract
By exploring the diversified compound with CaSR agonist activity, find that there is more excellent dense taste imparting effect, particularly there is the dense taste imparting effect of the high-titer of middle aftertaste type, and there is excellent stability, can the material of imparting body taste, providing package containing the dense taste imparting agent of this material, and comprises this material and other has the dense taste imparting agent of compound of the material of CaSR agonist activity.Comprise the dense taste imparting agent of γ-Glu-Nva-Gly (the positive valyl-glycine of L-gamma-glutamyl-L-), and this material and other are had the dense taste imparting agent of compound of combinations of substances of CaSR agonist activity.
Description
Technical field
The present invention relates to dense taste imparting agent and the dense taste imparting agent of compound of the peptide comprising display CaSR agonist activity.The invention still further relates to the food compositions of the peptide of the display CaSR agonist activity more than containing finite concentration.
Background technology
In recent years, along with the variation etc. of dietetic life, the requirement of consumer to the sense of taste improves day by day, the requirement of dense taste imparting agent of the excellence---cannot only expressed with 5 kinds of basic taste sensations such as sweet taste, saline taste, tart flavour, bitter taste and delicate flavours, but the sense of taste that mellow sense, full sense, be continuous sense and the edge sense of taste of coordinating the above-mentioned basic taste sensations such as sense are also strengthened---thereupon to giving " dense taste " also increases day by day.
In addition, calcium sensing receptor (CalciumSensingReceptor, CaSR), also calcium acceptor is called, the various intravital function of this receptor Signal Regulation, the material with CaSR agonist activity can be used as dense taste imparting agent (patent document 1 and 2, non-patent literature 3).
Multiple flavor pattern is there is, wherein to the dense taste of aftertaste type in can giving and higher dense taste imparting agent of tiring exists demand in above-mentioned " dense taste ".In addition, the material of imparting body taste is generally used for food etc., therefore needs it to have excellent stability.
Therefore, need to explore the diversified compound with CaSR agonist activity, to find that there is more excellent dense taste imparting effect, particularly there is the dense taste imparting effect of front taste type, and there is excellent stability, can the easy and material of imparting body taste produced at low cost, providing package containing this material dense taste imparting agent and this material and other are had the dense taste imparting agent of compound of combinations of substances of CaSR agonist activity.
In addition, for some γ-Glutamyl Peptides at N end with gamma-glutamyl amine, although the known example (patent document 3, non-patent literature 1,2) that it can be used as substrate to synthesize in enzymatic activity research etc., not yet knows the example and naturally occurring example that γ-Glu-Nva-Gly are actually used in food applications.
In addition, in above-mentioned patent document 1, describe γ-Glu-X-Gly (X is amino acid or amino acid derivativges) is the compound etc. with CaSR agonist activity, but for γ-Glu-Nva-Gly, the record not having actual synthesis in an embodiment and evaluate, not open particularly.In addition, the content of patent document 1 and 2 is incorporated to the record of this description.
Prior art document
Patent document
Patent document 1 International Publication No. 2007/055393 pamphlet
Patent document 2 International Publication No. 2008/139945 pamphlet
Patent document 3 International Publication No. 2007/066430 pamphlet
Non-patent literature
Non-patent literature 1MolecularPharmacology (1982), 21 (3), 629-36
Non-patent literature 2Biokhimiya (Moscow) (1972), 37 (4), 757-61
Non-patent literature 3TheJournalofBiologialChemistry, (2010), 285 (2), 1016-22
Summary of the invention
The object of the invention is by exploring the diversified compound with CaSR agonist activity, find that there is more excellent dense taste imparting effect, particularly there is the dense taste imparting effect of the high-titer of middle aftertaste type, and have excellent stability, can the material of imparting body taste, providing package containing this material dense taste imparting agent and this material and other are had the dense taste imparting agent of compound of combinations of substances of CaSR agonist activity.Further, object is to provide the food compositions containing this material certain density.
Inventors investigated diversified compound, result find surprisingly γ-Glu-Nva-Gly (the positive valyl-glycine of L-gamma-glutamyl-L-) have high CaSR agonist activity and very excellent dense taste give effect, particularly its flavor pattern can give in the dense taste of aftertaste type.Further discovery, what the γ-Glu-Nva-Gly found had more than 10 times compared with the γ-Glu-Val-Gly being similarly tripeptides highly tires, excellent in stability in addition, and the preferred flavor pattern that in display, aftertaste is stronger.Further γ-the Glu-Nva-Gly found can be used alone as dense taste imparting agent.Also find, by adding γ-Glu-Nva-Gly, the preferred food compositions that dense taste strengthens can be obtained.Also find to can be used as the dense taste imparting agent of compound this material and other with the combinations of substances of CaSR agonist activity, thus complete the present invention.
That is, the invention provides the dense taste imparting agent comprising γ-Glu-Nva-Gly.
In addition, the present invention also provides the food compositions containing γ-Glu-Nva-Gly (following, to be also called " food compositions of the present invention ").In addition, the invention provides (a) γ-Glu-Nva-Gly and (b) are selected from γ-Glu-X-Gly, γ-Glu-Val-Y, 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, the dense taste imparting agent of compound that combines of γ-Glu-Val-ol, γ-Glu-Ser, γ-Glu-Tau, γ-Glu-Cys (S-Me) (O), γ-Glu-Leu, one or more amino acid in γ-Glu-Ile, γ-Glu-t-Leu and γ-Glu-Cys (S-Me) or peptide, wherein X represents amino acid or amino acid derivativges, but is not Nva; Y represents amino acid or amino acid derivativges.
According to the present invention, the very powerful dense taste imparting effect having very excellent dense taste imparting effect, particularly have the middle aftertaste type of the uniqueness of flavor pattern general picture as shown in Figure 1 can be provided, and excellent in stability, can easy and the dense taste imparting agent produced at low cost and the dense taste imparting agent of compound.In addition, by the present invention, the certain density food compositions with the excellence of the material of dense taste imparting effect can be contained by providing package.
Use dense taste imparting agent of the present invention, the dense sense of the flavor can giving low-fat food as fat and lubrication sense, even if therefore reduce the fat content in fat-containing food, also can keep the dense sense same with former food, the food that health is highly profitable can be made.As this based food, can enumerate containing meat product and dairy products etc.Particularly tool has the following advantages: time on the feed containing the food of dense taste imparting agent of the present invention, be not at entrance at that time, but in the dense sense can felt afterwards as fat and lubrication sense.
Accompanying drawing is sketched
Fig. 1 shows the sense of taste general picture of the dense taste imparting agent of middle aftertaste type.
Detailed description of the invention
Comprise as the γ-Glu-Nva-Gly of object the positive valyl-glycine of L-gamma-glutamyl-L-and/or its salt, particularly edible salt that 3 amino acid are formed by peptide bond bonding in the present invention.
Give effect because γ-Glu-Nva-Gly has excellent dense taste, therefore can use as dense taste imparting agent.γ-Glu-Nva-Gly can be added, make relative to the weight of the food compositions of imparting body taste contain 0.1ppb ~ 99.9 quality %, preferably 1ppb ~ 10 quality %, more preferably 0.01ppm ~ 1 quality %, γ-Glu-Nva-Gly.That is, other embodiment of the present invention relates to the food compositions containing γ-Glu-Nva-Gly, the food compositions of the γ-Glu-Nva-Gly preferably containing 0.1ppb ~ 99.9 quality %.The food compositions of the ppm γ-Glu-Nva-Gly more preferably containing 0.01 ~ 50 weight.
In addition, by by dense taste imparting agent of the present invention, namely γ-Glu-Nva-Gly and other Seasoning Ingredients of at least one be selected from organic acid and the various yeast extracts etc. such as the inorganic salts such as nucleic acid, sodium chloride, citric acid such as the amino acidses such as sodium glutamate (MSG), inosinicacid (IMP) combinationally use, compared with situation about being used alone with other Seasoning Ingredients, the preferred flavoring that dense taste increases further can be provided.
For the present invention, " dense taste " (kokumi) refers to can not by five kinds of primary tastes, i.e. sweet taste (sweettaste), saline taste (saltytaste), tart flavour (sourtaste), the taste that bitter taste (bittertaste) and delicate flavour (umami) represent, more particularly not only make described primary taste strengthen, also make the mellow sense (thickness) of primary taste, full sense (growth) (profusely feeling (mounthfulness)), the taste that the edge taste (marginaltastes) such as sense (continuity) and coordination sense (harmony) that is continuous strengthens.At this, " imparting body taste " refers to five kinds of primary tastes that enhancing represents with sweet taste, saline taste, tart flavour, bitter taste and delicate flavour, and gives the edge taste with described primary tastes such as mellow sense, full sense, continuous sense and coordination sense.In addition, this also can be expressed as in flavour enhancement.Thus, dense taste imparting agent γ-Glu-Nva-Gly of the present invention also can be expressed as flavor reinforcing agent (flavorenhancer).Dense taste imparting agent γ-Glu-Nva-Gly of the present invention can also be used as sweetness enhancers, saline taste reinforcing agent, tart flavour reinforcing agent, bitter taste reinforcing agent or flavour enhancer.
In addition, the sense of taste is passed through along with the time after feed and changes, from after just feed, be called front taste (initialtaste), middle taste (middletaste) and aftertaste (aftertaste) in turn.These are relative concepts, and approximately, front taste, middle taste and aftertaste are respectively feed latter 0 to 2 second, the flavor felt after 2 to 5 seconds and 5 seconds.In addition, within 0 to 5 second, being called " front middle taste ", about about 30 seconds was extremely " middle aftertaste " (see Fig. 1) after 2 seconds.Due to for the evaluation be divided three classes, enter trencherman and be difficult to be absorbed in evaluation, therefore the general conventional evaluation being divided into two classes.
The material with CaSR activity confirms by methods such as human body taste tests for the effect of dense taste and flavor pattern.It can the test shown in the embodiment of description of the present invention be example that this kind of human body sense of taste official can test, but is not limited to this.
In this manual, so-called " CaSR ", mean calcium sensing receptor (CalciumSensingReceptor), it belongs to the C class of seven transmembrane receptors, also referred to as calcium acceptor.In this manual, so-called " CaSR activator " means to be combined with above-mentioned CaSR, activates the material of CaSR.In addition, in this manual, so-called " activating CaSR ", means part and is combined with CaSR, activate guanine nucleotide binding protein matter and carry out intracellular signaling.In addition, be combined with CaSR and the character that activates CaSR is called that " CaSR activator " is active.
Show that screening has the method for the compound of CaSR agonist activity below particularly, but be not limited to these steps.
1) tested substance being added into the CaSR activity determining system for measuring CaSR activity, measuring CaSR active.
2) by active compared with CaSR activity when not adding tested substance for CaSR when adding tested substance.
3) tested substance showing CaSR agonist activity when adding tested substance is selected.
The mensuration of CaSR activity such as can utilize and use the Analytical system of the cell of expressing CaSR to carry out.Above-mentioned cell can be the cell of endogenous expression CaSR, also can be the recombinant cell that outer seedbed has imported CaSR gene.For above-mentioned CaSR activity determining system, as long as can detect the combination (reaction) of activated material and CaSR when adding, to CaSR, there is specific extracellular ligand (activated material) to the cell of above-mentioned expression CaSR, or the combination (reaction) of response activated material and CaSR and conduct in cell detectable signal, can use with no particular limitation.When being detected that by the reaction with tested substance CaSR is active, judge that this tested substance has CaSR stimulating activity.
Above-mentioned CaSR can preference as the people CaSR of the people CaSR coded by said gene logged in GenBank accession number NM_000388.And, CaSR is not limited to the protein by the gene code of above-mentioned sequence, can be have more than 60%, preferably more than 80% by with above-mentioned sequence, the more preferably protein of coded by said gene of homogeneity of more than 90%, as long as described gene code has the protein of CaSR function.In addition, the function of CaSR by by these genes at cells, and measure the change of electric current or the change of intracellular calcium concentration when adding calcium and investigated.
There is no particular restriction in the source of above-mentioned CaSR, is not only above-mentioned people CaSR, also can enumerates the CaSR comprising all animal origins such as mouse, rat and dog.
As mentioned above, CaSR activity can utilize the living cells of expressing CaSR or its fragment, and express the cell membrane of CaSR or its fragment, the vitro system etc. comprising the protein of CaSR or its fragment confirms.
Below show the example using living cells, but be not limited to this.
By CaSR at Xenopus Oocytes, express in the cultured cell such as hamster ovary cell and foetal human kidney cells.This can by by CaSR gene clone in the plasmid keeping foreign gene, then it is imported with the form of plasmid or imports with it as the cRNA of template and realizing.The detection of reaction can use the fluorescence indicator of electrophysiology means and intracellular Ca2+ rising.
First the expression of CaSR is confirmed by the response caused by calcium or specific activation agent.Use the calcium for about 5mM concentration can detect that the egg mother cell of endocellular electricity stream maybe can detect the cultured cell of the fluorescence of fluorescence indicator.Change calcium concentration and measure concentration dependent.Afterwards, tested substance is adjusted to 1 μM ~ about 1mM, adds egg mother cell or cultured cell, under above-mentioned tested substance exists, measure CaSR activity, measure the CaSR agonist activity of above-mentioned tested substance thus.
In addition, more specifically, the test example that the test of CaSR agonist activity can enumerate present specification is example, but is not limited thereto.
The amino acid combined with γ-Glu-Nva-Gly in the dense taste imparting agent of compound of the present invention or peptide are for being selected from by γ-Glu-X-Gly (X represents amino acid except Nva or amino acid derivativges), γ-Glu-Val-Y (Y represents amino acid or amino acid derivativges), 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, one or more amino acid in the group that γ-Glu-t-Leu and γ-Glu-Cys (S-Me) forms or peptide, at this, so-called amino acid comprises Gly, Ala, Val, Leu, Ile, Ser, Thr, Cys, Met, Asn, Gln, Pro, Hyp, the neutral amino acids such as t-Leu, Asp, the acidic amino acids such as Glu, Lys, Arg, the basic amino acids such as His, Phe, Tyr, the aromatic amino acids such as Trp, also comprise homoserine, citrulling, ornithine, butyrine, norvaline, nor-leucine and taurine etc.In addition, also can be Terleu (tert-leucine), cycloleucine (cycloleucine), α-aminoacid, non-natural (nonprotein constitutive character) amino acid such as L-penicillamine.And in peptide γ-Glu-X-Gly, X can be amino acid or derivatives thereof as above arbitrarily, is preferably the amino acid or derivatives thereof beyond Cys.Wherein, as the peptide combinationally used, preferred γ-Glu-Val-Gly, γ-Glu-Abu-Gly, γ-Glu-tLeu-Gly, γ-Glu-Nva and γ-Glu-Abu etc.
Specifically, dense taste imparting agent of the present invention comprises γ-Glu-Nva-Gly, there is the dense taste imparting effect of the excellence of the middle aftertaste type of the uniqueness of general picture as shown in Figure 1, therefore preferred by itself and the peptide with the general picture being different from such general picture, such as, the γ-Glu-Abu-Gly be preferably inclined to front taste, γ-Glu-Abu etc. combinationally use.
In this manual, the abbreviation of amino acid residue means following amino acid:
(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: methyl amimoacetic acid
(23) Cit: citrulling
(24) N-Val (or Nva): norvaline (2-aminovaleric acid)
(25) N-Leu (or Nle): nor-leucine
(26) Abu: butyrine
(27) Tau: taurine
(28) Hyp: hydroxyproline
(29) t-Leu: Terleu
(30) Cle: cycloleucine
(31) Aib: α-aminoacid (α-aminoisobutyricacid, 2-methylalanine)
(32) Pen:L-penicillamine (penicillamine)
(33) allo-Thr: allothreonine
(34) allo-Ile: alloisoleucine
The amino acid side chains such as in addition, so-called " amino acid derivativges " is above-mentioned amino acid whose various derivative, the sulfydryl of such as special acid and alpha-non-natural amino acid, amino alcohol or terminal carbonyl or amino, cysteine are by various substituting group substituent.So-called substituting group can enumerate alkyl, acyl group, hydroxyl, amino, alkyl amino (alkylamino), nitro, sulfonyl and various protecting groups etc., such as, comprises Arg (NO
2): N-γ-Nitro-Arginine, Cys (SNO): S-nitro cysteine, Cys (S-Me): S-methyl cysteine, Cys (S-pi-allyl): S-allyl cysteine, Val-NH
2: valine amide (valinamide), Val-ol: valerian ammonia alcohol (valinol) (2-amino-3-methyl-1-butanol) etc.And, in this manual, γ-Glu-Cys (SNO)-Gly has following structural formula, and (O) in above-mentioned γ-Glu-Met (O) and γ-Glu-Cys (S-Me) (O) formula means sulfoxide structure.(γ) of so-called γ-Glu is meant glutamic acid and is combined with other amino acid by the carboxyl of γ position.
S-ニトロソグルタチオン(S-Nitrosoglutathione(GNSO))
GSNO (GNSO)
γ-Glu-Nva-Gly of the present invention and with the amino acid of its combination or peptide, commercially available product can be used when having commercially available, in addition, suitably can use and be obtained by known approaches such as enzyme reaction synthetic methods via (1) chemical synthesis process or (2), chemical synthesis is easier.Total number of atnino acid contained by the γ-Glu-Nva-Gly that uses in the present invention is 3 residues, and length is shorter, and chemical synthesis process is easy, industrially highly beneficial.In addition, at γ-Glu-Nva-Gly of the present invention and be chemical synthesis with the amino acid of its combination or peptide, undertaken by using peptide synthesizer synthesis or this oligopeptides semi-synthetic.It is example that chemical synthesis process can enumerate peptide solid-phase synthesis etc.The peptide so synthesized such as, by common means, ion-exchange chromatography, and RPLC, affinity chromatography etc. carry out purifying.Such peptide solid-phase synthesis, and peptide purification is afterwards well known in the art.
In addition, the γ-Glu-Nva-Gly used in the present invention and with the amino acid of its combination or peptide be produced by enzyme reaction when, also can use the method that such as No. WO2004/011653, International Publication pamphlet is recorded.Namely; can by by esterified for the c-terminus of a side or amidated amino acid or dipeptides and amino acid be the amino acid of free state (such as; the shielded amino acid of carboxyl) react under the existence of peptide-forming enzyme, and by the dipeptides of generation or three peptide purifications, produced.So-called peptide-forming enzyme, can enumerate the culture of the microorganism with the ability generating peptide, the microbial cells be separated by this culture, or the bacterial disposing thing of this microorganism, or from the peptide-forming enzyme of this microorganism.And, the item that No. WO2004/011653 is recorded is incorporated to the record of this description.
Further, except enzyme method as above and chemical synthesis process, the peptide used in the present invention also can be present in the plant such as vegetables and fruit, the microorganisms such as yeast, and in other natural materials.In naturally occurring situation, also can extract rear use from them.
Dense taste imparting agent of the present invention and the dense taste imparting agent of compound can individually, or to mix with acceptable carrying body or other Seasoning Ingredients with diet product and be used as flavoring.For other Seasoning Ingredients, can enumerate the organic acids such as the inorganic salts such as nucleic acid, sodium chloride, citric acid such as amino acids, inosinicacid (IMP) such as spices, carbohydrate, sweetener, dietary fiber, vitamin, sodium glutamate (MSG), and various yeast extract is example.
And, as the preferred low-fat food of food compositions comprising dense taste imparting agent of the present invention or compound dense taste imparting agent, be the food originally containing fat, particularly this fat content food through reducing.At this, " " have identical meanings, comprise solid and liquid, can be any fat in animal fat and vegetablefats to fatty "AND" grease.
This type of low-fat food, milk, Yoghourt, butter, milk wet goods dairy products can be enumerated, margarine, coffee milk, sauce (sauce), roux (roux) etc. contain the food of animal fat and/or vegetable fat, the emulsified foods etc. such as salad dressing (dressing), mayonnaise (mayonnaise), containing cooked meat various currys, stew, various soup etc. containing meat extract.In addition, also can enumerate the beefsteak be made up of cooked low fat beef or barbecue class etc., commonly cure little food without fried process.Wherein, as low-fat food, preferably usually food fat content 1/2 ~ 1/3.
By making above-mentioned low-fat food contain dense taste imparting agent of the present invention, when these food edible, not initial but can in the dense sense next experienced as fat and lubrication sense.
It should be noted that, about milk or Yoghourt, the fat of usual product is 3 ~ 4%, but also there will be a known zero fat products (fat about 0.1%).Dense taste imparting agent of the present invention or compound dense taste imparting agent are also effective for these zero fat products.
In addition, dense taste imparting agent of the present invention or the dense taste imparting agent of compound are especially preferably added into the food containing pork source.That is, the present invention also provides the food compositions containing γ-Glu-Nva-Gly and pork source.For the food containing pork source, be not particularly limited, can enumerate such as: pork extract, sausage, i.e. noodle soup (and Noodles ス mono-プ) etc.Content for pork source is not particularly limited, and can enumerate is such as 0.005 ~ 80 about % by weight in food compositions of the present invention.
In addition, dense taste imparting agent of the present invention or the dense taste imparting agent of compound are also preferably added into the food containing beef source.That is, the present invention also provides the food compositions containing γ-Glu-Nva-Gly and beef source.For the food containing beef source, be not particularly limited, can enumerate such as: the sauce etc. of beef extract, the Salted beef that salts down (コ mono-Application PVC one Off), the soup using beef, use beef.Content for beef source has no particular limits, and can enumerate is such as 0.005 ~ 80 about % by weight in food compositions.
γ-the Glu-Nva-Gly used in the present invention and the amino acid combinationally used or peptide also comprise the form of salt.The occasion of salt form can be formed at γ-Glu-Nva-Gly of the present invention and the amino acid combinationally used or peptide, as long as pharmaceutically acceptable, the edible salt of this salt, such as, for acidic-groups such as carboxyls, ammonium salt can be enumerated, salt alkali-metal with sodium, potassium etc., with the salt of the alkaline-earth metal such as calcium, magnesium, aluminium salt, zinc salt, with the salt of the organic amines such as triethylamine, monoethanolamine, morpholine, pyrrolidines, piperidines, piperazine and dicyclohexyl amine, with the salt of the basic amino acid such as arginine, lysine.In addition, for basic group, can enumerate and hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, the salt of the inorganic acids such as hydrobromic acid, with acetic acid, citric acid, benzoic acid, maleic acid, fumaric acid, tartaric acid, butanedioic acid, tannic acid, butyric acid, 2-(4-(2-hydroxybenzoyl)) benzoic acid (hibenzicacid), flutter acid (pamoicacid), enanthic acid (enanthicacid), capric acid (decanoicacid), 8-Chlorotheophyline (acid of テ オ Network Le), salicylic acid, lactic acid, oxalic acid, the salt of the organic carboxyl acid such as mandelic acid and malic acid, and with methanesulfonic acid, benzene sulfonic acid, the salt of the organic sulfonic acids such as p-methyl benzenesulfonic acid.
Dense taste imparting agent of the present invention, food compositions or the dense taste imparting agent of compound can with dried powders, and paste, any form such as solution is used, and does not limit its physical property.
Dense taste imparting agent of the present invention, food compositions or the dense taste imparting agent of compound can be coupled in the various diet product such as food, beverage, flavoring and use.
By dense taste imparting agent of the present invention, food compositions or the dense taste imparting agent of compound are coupled to food, beverage, when using in the various diet product such as flavoring, final γ-Glu-Nva-GLy is measured and the amino acid that combinationally uses or peptide amount for, as long as the amount of desired effect can be obtained, without particular limitation, as the amount of γ-Glu-Nva-Gly and/or the amount of amino acid or peptide, with food, the gross mass of beverage or flavoring etc. is benchmark, be 0.1ppb ~ 99.9 quality % separately, preferred 1ppb ~ 10 quality %, more preferably 0.01ppm ~ 1 about quality %.
In the various diet product such as food, beverage, flavoring being combined with dense taste imparting agent of the present invention, food compositions or the dense taste imparting agent of compound, also can coordinate the acceptable any solid of diet product or liquid carrying body, suitable Seasoning Ingredients etc. further.
As above-mentioned carrying body, such as glucose, lactose, sucrose, starch, sweet mellow wine, dextrin, fatty glyceride, polyethylene glycol, HES, ethylene glycol, Polyoxyethylene Sorbitan acid esters, gelatin, albumin, amino acid, water and physiological saline etc. can be enumerated.
Above-mentioned Seasoning Ingredients can be the Seasoning Ingredients that any this area uses, and is not particularly limited, more specifically can enumerates person mentioned above.
Above-mentioned carrying body, other Seasoning Ingredients etc., its content is all without particular limitation.
In above-mentioned Seasoning Ingredients, yeast extract can use any yeast extract, to thalline, its condition of culture in its source, extraction process method etc. is all not particularly limited, also further across the yeast extract of heat treatment, ferment treatment, concentrated, powdered process etc.
The present invention also provides the manufacture method of various diet product, it is characterized in that, adds γ-Glu-Nva-Gly and make it contain 1 quality ppb ~ 99.9 quality % in the intermediate products of various diet product.In this as various diet product, preferred low-fat food.
The present invention also provides the manufacture method of various diet product, it is characterized in that, food compositions of the present invention in the intermediate products of various diet product.In this as various diet product, preferred low-fat food.
With regard to using the intermediate products manufacture method of dense taste imparting agent of the present invention, be preferably as follows the manufacture method of the intermediate products of diet product or diet product, the method comprises: the flavor reinforcing agent comprising γ-Glu-Nva-Gly is added being mixed to drink raw-food material (such as, delicate flavour raw material, protein hydrolysate, poultry meat extract) step, and as required, cook the step of the diet product raw mixture obtained further.
At this, the flavor reinforcing agent comprising γ-Glu-Nva-Gly is added the step be mixed in drink raw-food material preferably include: the γ-Glu-Nva-Gly concentration making the intermediate products of diet product is 0.01 ~ 999900 weight ppm, is preferably the step of 0.1 ~ 200000 weight ppm.
In addition, also preferably include further: the intermediate products of diet product are added into other drink raw-food material (such as, agricultural product, aquatic products, poultry meat, dairy products or its processed food etc.), γ-Glu-Nva-Gly the concentration making the diet product of acquisition is 0.01 ~ 50 weight ppm, is preferably the step of 0.05 ~ 20 weight ppm.
In addition, the flavor reinforcing agent comprising γ-Glu-Nva-Gly is added the step being mixed to drink raw-food material to preferably include: the γ-Glu-Nva-Gly concentration making diet product is 0.01 ~ 50 weight ppm, is preferably 0.05 ~ 20 weight ppm.
In above-mentioned manufacture method, preferred diet product are the food containing pork source.Now, preferably containing the γ-Glu-Nva-Gly of 0.01 ~ 50 weight ppm, the pork source of 0.005 ~ 80 % by weight and other raw-food material.
In above-mentioned manufacture method, preferred diet product are the food containing beef source.Now, preferably containing the γ-Glu-Nva-Gly of 0.01 ~ 50 weight ppm, the beef source of 0.005 ~ 80 % by weight and other raw-food material.
In addition, as object food of the present invention, except above-mentioned food, the food (sweet food based food) such as Western-style pastry and dessert based on sweet taste such as ice cream, honey, organe peel jam and strawberry jam also can be enumerated as preferred food, and the food (appetizer based food) such as processed food, dish to go with rice, steamed buns etc, little food based on saline taste such as chicken soup.
Below enumerate embodiment to be illustrated the present invention is more detailed, but these do not limit the present invention.
Embodiment
The synthesis of (synthesis example 1) γ-Glu-Nva-Gly (the positive valyl-glycine of γ-L-glutamy-L-)
Boc-Nva (tert-butoxycarbonyl-L-norvaline, 4.44g, 20.4mmol) and Gly-OBzlHCl (glycine benzyl hydrochloride, 4.12g, 20.4mmol) is dissolved in carrene (CH
2cl
2, 100ml) in.Reactant liquor is kept 0 DEG C, with the addition of triethylamine (Et
3n, 3.13ml, 1.1 equivalents, 22.4mmol), HOBtH
2o (I-hydroxybenzotriazole hydrate, 3.44g, 1.1 equivalents, 22.4mmol) and WSCHCl (1-ethyl-3-(3-dimethylaminopropyl)-phosphinylidyne diimmonium salt hydrochlorate, 4.30g, 1.1 equivalents, 22.4mmol).The temperature of reactant liquor is slowly raised, at room temperature stirs and spend the night (16 hours).By reactant liquor reduced pressure concentration, ethyl acetate (150ml) is added in residue, organic layer is proceeded as follows: utilize water (50ml), 5% aqueous citric acid solution (50ml) cleans 2 times and utilize saturated aqueous common salt (50ml), 5% sodium bicarbonate aqueous solution (50ml) cleans 2 times and utilize saturated aqueous common salt (50ml) to clean, and utilize anhydrous magnesium sulfate drying.Cross and filter magnesium sulfate, by filtrate reduced in volume.Produce crystallization when adding n-hexane in residue, filter and collect crystallization, carrying out drying under reduced pressure and obtain Boc-Nva-Gly-OBzl crystallization (6.88g, 18.9mmol).
4NHCl/ bis-is added in Boc-Nva-Gly-OBzl (6.88g, 18.9mmol)
alkane solution (94.5ml), at room temperature stirs 1 hour.Reduced pressure concentration removing two
alkane, adds n-hexane (30ml) and carries out reduced pressure concentration, repeat 3 times, obtain H-Nva-Gly-OBzlHCl with quantitative yield in residue.
H-Nva-Gly-OBzlHCl is dissolved in carrene (130ml), reactant liquor is remained on 0 DEG C.Z-Glu-OBzl (N-α-benzyloxycarbonyl group-Pidolidone α-benzyl ester (N-α-Carbobenzoxy-L-glutamicacid α-benzylester), 7.03g, 18.9mmol), triethylamine (2.90ml, 1.1 equivalents, 20.8mmol), HOBtH is added in reactant liquor
2o (3.20g, 1.1 equivalents, 20.8mmol) and WSCHCl (3.98g, 1.1 equivalents, 20.8mmol).The temperature of reactant liquor is slowly raised, at room temperature stirs and spend the night (16 hours).Reduced pressure concentration reactant mixture, ethyl acetate (1000ml) is added in residue, organic layer is proceeded as follows: utilize water (100ml), 5% aqueous citric acid solution (100ml) clean 2 times, utilize saturated aqueous common salt (100ml), 5% sodium bicarbonate aqueous solution (100ml) cleans 2 times and utilizes saturated aqueous common salt (100ml) to clean, and utilizing anhydrous magnesium sulfate to carry out drying.After solution being heated to 50 DEG C, crossing and filter magnesium sulfate, reduced pressure concentration is carried out to filtrate.Add n-hexane when just starting crystallization, crystallization is separated out fully.Filter and collect crystal, carrying out drying under reduced pressure and obtain crystal Z-Glu (Nva-Gly-OBzl)-OBzl (10.16g, 16.4mmol).
Z-Glu (Nva-Gly-OBzl)-OBzl (10.16g, 16.4mmol) and 5% palladium carbon (5% palladium/carbon, 1.20g) is added in the mixed liquor of ethanol (250ml) and water (30ml), 50 DEG C spend the night (14 hours), carry out contact reduction under a hydrogen atmosphere.In reaction, a little adds water (100ml).Cross and filter palladium carbon, by filtrate reduced in volume.Residue is recrystallized from a small amount of water and ethanol and obtains white crystal γ-Glu-Nva-Gly (4.59g, 15.1mmol).Its characteristic value is as follows.
ESI-MS:(M+H)
+=304.1
1h-NMR (400MHz, D
2o) δ (ppm): 0.82 (3H, t, J=7.4Hz), 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 8.9Hz)
The preparation of (test example 1) CaSR expression plasmid
Being prepared as follows of CaSR expression plasmid is carried out.
With the DNA sequence dna logged in NCBI (CaSR (calcium acceptor): NM_000388, SEQIDNO:1,2) based on, PCR is used to synthesize few DNA (forward primer (SEQIDNO:3:ACTAATACGACTCACTATAGGGACCATGGCATTTTATAGCTGCTGC TGG), and reverse primer (SEQIDNO:4:TTATGAATTCACTACGTTTTCTGTAACAG).
With the cDNA (Clontech Inc.) in people's kidney source for material, use aforementioned primer, and PfuUltraDNA polymerase (Stratagene Inc.), implement PCR with following condition.94 DEG C after 3 minutes, 94 DEG C 30 seconds, 55 DEG C 30 seconds, 72 DEG C circulations in 2 minutes 35 times, then 72 DEG C are reacted for 7 minutes.Carry out agarose electrophoresis, after with DNA staining reagent, detect whether increased by PCR by Ultraviolet radiation.In addition, compared with being marked by the DNA molecular amount known with the size of carrying out electrophoresis simultaneously, the chain length of PCR primer is confirmed.
Plasmid vector pBR322 Restriction Enzyme EcoRV (Takara Inc.) is cut off, connects the genetic fragment by pcr amplification gained in this cut-out site with coupled reaction kit (Promega Inc.).With this reaction solution transformation of E. coli DH5 α strain, screening remains with the transformant of the plasmid of wherein having cloned pcr amplification product, further by DNA base sequence analysis confirmation pcr amplification product.
This kind of recombinant plasmid is used to prepare people CaSR expression plasmid hCaSR/pcDNA3.1.
The evaluation of (test example 2) CaSR agonist activity
293E cell (is expressed the HEK293 cell of EBNA1, ATCCNo.CRL-10852) under the G418 (Geneticin) of 200 μ g/ml exists, in the middle cultivation of the DMEM/Ham's-F12 (Dulbecco'smodifiedEaglemedium, Nakaraitesk containing 3.15/ml glucose) of the hyclone comprising 10%.With 3 × 10
6cell/10ml is inoculated in F25 flask, at CO
2incubator (5%CO
2, 37 DEG C) in leave standstill after 24 hours, with transfection agents Fugene6 (Roche) transfected with human CaSR expression plasmid hCaSR/pcDNA3.1.At CO
2after placing 6 ~ 7 hours in incubator, reclaim cell with the DMEM/Ham's-F12 comprising 10% hyclone, be inoculated in 96 orifice plates (BD-Biocoat) of poly-D-Lys bag quilt with 70000 cells/well.
At CO
2after leaving standstill 24 hours in incubator, remove culture medium from 96 orifice plates of this cell of inoculation, will be dissolved in and measure buffer solution (146mMNaCl, 5mMKCl, 1mMMgSO
4, 1mg/ml glucose, 20mMHEPES (pH7.2), 0.75 ~ 1.25mMCaCl
2) Ca
2+fluorescence indicator Calcium4AssayKit (MolecularDevices) adds with 200 μ l/ holes, at 37 DEG C, leave standstill 1 hour, then at room temperature leaves standstill 10 minutes, makes it take in indicator.
Add the test compound of the mensuration buffer solution be dissolved in containing 0.1%BSA to this 96 orifice plate with 50 μ l/ holes, in 3 minutes, measure the change of fluorescence intensity with FLEXStation (MolecularDevices).
(EC
50computational methods)
The maximum of fluorescence intensity before and after being added by the automatic calculating deterministic compound of FLEXStation and the difference (RFU (Max-Min)) of minimum of a value.RFU (Max-Min) when adding compound with Cmax is defined as 100%, the RFU (Max-Min) during the mensuration buffer solution containing 0.1%BSA used not containing test compound is defined as 0% and carrys out calculated activity rate.Carried out curve fitting by table software for calculation Xfit or Graph-Pad-Prism, determine compound concentration EC during activity rate 50%
50value.Result is shown in table 1.In addition, the data of the tripeptides measured equally as comparative example are as shown in table 2, table 3.It should be noted that, the data recorded in table 3 are the data disclosed in non-patent literature 3.
Table 1
| Compound | EC50，μM |
| γ-Glu-Nva-Gly | 0.055 |
| γ-Glu-Val-Gly | 0.075 |
Table 2
| Compound | EC50，μM |
| γ-Glu-Ala-Gly | 0.016 |
| γ-Glu-tLeu-Gly | 0.09 |
| γ-Glu-Thr-Gly | 2.8 |
Table 3
| Compound | EC50，μM |
| γ-Glu-Val-Gly | 0.039 |
| γ-Glu-Abu-Gly | 0.025 |
| γ-Glu-Cys-Gly | 0.7 |
Unexpectedly found by the above results, the high CaSR that γ-Glu-Nva-Gly has the peptide same degree with other with γ-Glu-X-Gly type structure is active.
the dense taste of embodiment 1 gives active evaluation
For γ-Glu-Nva-Gly, checked dense taste by quantitative sense evaluation experimental and give active intensity.
Quantitative sense evaluation experimental is implemented as follows.Containing sodium glutamate (0.05g/dl), inosinicacid monophosphate (acid of イ ノ シ Application acid one リ Application) (0.05g/dl), in the distilled water of sodium chloride (0.5g/dl), using as under the condition that the compounds of sample mixes with 0.000001 ~ 0.1g/dl, measure dense taste and give active intensity.For the sample of contrast in acidity relatively without interpolation after test portion dissolving, be adjusted to by NaOH and relatively used without the scope of contrast pH ± 0.2 of interpolation.Sense scoring is contrast: 0 point, strong: 3 points, and very by force: 5 points, simultaneously in order to make yardstick definitely, by taste in before γ-Glu-Cys-Gly, aftertaste respectively provides is 3.0 points.Scoring uses linear scale method, more particularly, with for the straight line of position indicating-5 ~ 0 ~ 5 points, using the method charged to as position of marking accordingly.In addition, allow the experience of accumulative tool above foodstuff flavouring exploitation one year, can judge that the difference of tiring of γ-Glu-Cys-Gly and the γ-Glu-Val-Gly added in delicate flavour saline taste solution is that people's (regularly confirming) of about 10 times is as judging panel.Evaluation ideas n=4 implements.In addition, " front middle taste " refers to the flavor of after keeping in the mouth 0 ~ 5 second, and aftertaste is flavor after this.The dense taste showing wide region by inspection compound in above-mentioned interpolation concentration is given active, but only the result of representational concentration is shown in table 4.
This result shows, and except the tripeptides except γ-Glu-Nva-Gly, all demonstrates the activity of about about 10 times that are glutathione (γ-Glu-Cys-Gly) to the maximum, and surprisingly, γ-Glu-Nva-Gly shows higher high activity, more than 100 times.
Table 4
Known γ-Glu-Nva-Gly dense taste compared with γ-Glu-Cys-Gly give active improves about 100 times, compared with γ-Glu-Val-Gly etc., dense taste imparting activity is at least high about 10 times.In addition, the same with γ-Glu-Ala-Gly, γ-Glu-Abu-Gly, γ-Glu-tLeu-Gly, do not have peculiar smell (the convergence taste etc. of aftertaste), visible γ-Glu-Nva-Gly is excellent.
the dense taste of embodiment 2 gives active evaluation
For γ-Glu-Nva-Gly, in order to clear and definite its is middle aftertaste type, give active intensity by the quantitative sense evaluation test in other assessment item to dense taste and be studied.
Quantitative sense evaluation experimental is implemented as follows.For ease of aftertaste in distinguishing, evaluating liquid does not use inosinicacid monophosphate, with the delicate flavour after in reducing.That is, in the distilled water containing sodium glutamate (0.1g/dl), sodium chloride (0.4g/dl) using when mixing as the compounds of sample with 0.000001 ~ 0.1g/dl, measure dense taste and give active intensity.For the sample relative to the contrast without interpolation being acidity after sample dissolving, the scope being adjusted to relative contrast pH ± 0.2 without interpolation with NaOH uses.Sense scoring is: contrast: 0 point, strong: 3 points, very by force: 5 points, simultaneously in order to make yardstick definitely, using taste, middle aftertaste before γ-Glu-Cys-Gly as 3.0 points.Scoring uses linear scale method, more particularly, with for the straight line of position indicating-5 ~ 0 ~ 5 points, using the method charged to as position of marking accordingly.In addition, allow the experience of accumulative tool above foodstuff flavouring exploitation one year, can judge that the difference of tiring of γ-Glu-Cys-Gly and the γ-Glu-Val-Gly added in delicate flavour saline taste solution is that people's (regularly confirming) of about 10 times is as judging panel.Evaluation ideas n=4 implements.In addition, " front taste " refers to the flavor of after keeping in the mouth 0 ~ 2 second, middle aftertaste be this after flavor.The dense taste showing wide region by inspection compound in above-mentioned interpolation concentration is given active, but only the result of representational concentration is shown in table 5.
This result also shows, and the activity of γ-Glu-Val-Gly is about about 10 times of glutathione (γ-Glu-Cys-Gly), and the activity of γ-Glu-Nva-Gly improves further, has the high activity more than 100 times.
Table 5
Known γ-Glu-Nva-Gly has excellent dense taste and gives active, but also has the advantage such as speciality, free from extraneous odour (convergence taste etc.) that flavor pattern has middle aftertaste.In addition, γ-Glu-Nva-Gly dense taste compared with γ-Glu-Cys-Gly gives active height about 100 times, compared with γ-Glu-Val-Gly etc., and it is at least high about 10 times that dense taste gives activity, therefore can use at much lower concentrations.Therefore, can be easier and dense taste imparting agent is provided at low cost, be also industrially very favorable.
in embodiment 3 food, dense taste gives active evaluation
By sense evaluation test, for γ-Glu-Nva-Gly when being actually used in food compared with the γ-Glu-Val-Gly of high-titer effect whether obtain extremely improving being studied.
Sense evaluation test is implemented as follows.For thinking the food that middle aftertaste is strong, being the representative of the main food (sweet food based food) such as Western-style pastry, dessert as sweet taste, employing commercially available ice cream, honey, organe peel jam and strawberry jam.As the representative that saline taste is the main food (appetizer based food) such as processed food, dish to go with rice, steamed buns etc, little food, use in commercially available chicken soup, mashed potatoes add 0.1 % by weight commercial powder pepper and add in the paste obtained, commercially available Jiang Rong and mashed potatoes 2 % by weight butter and the paste that obtains.Combined amount for the γ-Glu-Val-Gly compared is 0.002 % by weight, definite effect under this combined amount.For γ-Glu-Nva-Gly, determine with the enhancing of 0.0000001 ~ 0.01 % by weight mixed flavor entirety (dense taste gives active intensity).Contrast is the food without interpolation.For sense scoring, in order to the difference not producing decimal point waits fine difference, provide as follows: contrast: ±, slightly by force :+, strong: ++, very strong: +++.To+be evaluated as 1 point, ++ be 2 points, +++ be 3 points, if mean values is such as 2.2, be then rounded to 2=++, like this.In addition, allow the experience of accumulative tool above foodstuff flavouring exploitation one year, can judge that the difference of tiring of γ-Glu-Cys-Gly and the γ-Glu-Val-Gly added in delicate flavour saline taste solution is that people's (regularly confirming) of about 10 times is as judging panel.Evaluation ideas n=4 implements.With regard to γ-Glu-Val-Gly, the dense taste showing wide region in above-mentioned interpolation concentration is given active, but the concentration results that can clearly compare is as shown in table 7 and table 8.
Also show according to this result, relative to have about 10 times to glutathione (γ-Glu-Cys-Gly) dense taste give active γ-Glu-Val-Gly, γ-Glu-Nva-Gly display further has the high activity of more than 5 ~ 13 times.
Table 7
Table 8
With regard to γ-Glu-Nva-Gly, known its at the appetizer based food from reality in all food being feature with middle aftertaste in sweet food based food, all there is the likely dense taste improving overall flavor and give active.Show in addition, γ-Glu-Nva-Gly is in the food of reality, and compared with γ-Glu-Val-Gly, the significantly high dense taste with more than 5 ~ 13 times is given active.Therefore, when hope improves, but due to the reason of quality stability cannot coordinate more raw material time, by using γ-Glu-Nva-Gly, get final product oil recovery enhancement with atomic little consumption.In addition, dense taste imparting agent can be provided at low cost.
embodiment 4 γ-Glu-Nva-Gly gives the effect of pork extract
Find to compare with γ-Glu-Val-Gly with γ-Glu-Cys-Gly (glutathione), more the time of morning starts to strengthen the dense taste imparting efficacy activities of γ-Glu-Nva-Gly after consumption.Thus, by sense evaluation test confirm γ-Glu-Nva-Gly for be not completely in taste type, but the pork extract that flavor strengthens a little earlier has effect more more significant than γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly.
Sense evaluation test is implemented as follows.Marketing pork extract (solid constituent 55.1 % by weight, salinity 9.3 % by weight) is dissolved and is 5.0 % by weight in the hot water, be mixed with pork extract solution.For this pork extract solution, mix γ-Glu-Nva-Gly, the γ-Glu-Cys-Gly as sample or γ-Glu-Val-Gly.Measure and use paired comparison test's method, evaluation is compared: the γ-Glu-Nva-Gly of (1) 0.0003 % by weight gives the γ-Glu-Cys-Gly of 0.02 % by weight of activity with having equal dense taste to following, γ-the Glu-Val-Gly of the γ-Glu-Nva-Gly of (2) 0.0003 % by weight and 0.0003 % by weight of isodose, γ-the Glu-Nva-Gly of (3) 0.0003 % by weight gives the γ-Glu-Val-Gly of 0.002 % by weight of activity with having equal dense taste, and judge that whichever is " under the condition of the balance of the flavor and local flavor that do not change pork extract, strengthen its flavor and local flavor by judging panel, preferred ".Evaluate and implement with n=9.The judging panel's number γ-Glu-Nva-Gly of 0.0003 % by weight being evaluated as " strengthening its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change pork extract, is preferred " is as shown in table 9.
Visible according to this result, even under the condition that the equal dense taste such as shown in (1) and (3) is tired, γ-Glu-Nva-Gly also obviously " under the condition of the balance of the flavor and local flavor that do not change pork extract, strengthen its flavor and local flavor, be preferred ".
Table 9
N=9
*)in the level of signifiance of 5%, can think that γ-Glu-Nva-Gly strengthens its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change pork extract, be preferred
Known with have compared with γ-Glu-Cys-Gly, γ-Glu-Val-Gly that equal dense taste gives under active concentration, the effect that γ-Glu-Nva-Gly has " strengthening its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change pork extract, is preferred " very gives prominence to like this.Pork source is worldwide widely used in flavoring, soup, poultry meat processed goods, the cooking processed goods, dessert, little food etc.
Therefore, γ-Glu-Nva-Gly can with more low cost, and small consumption improves food, is also industrially very favorable.
embodiment 5 γ-Glu-Nva-Gly gives the effect of beef extract
Find to compare with γ-Glu-Val-Gly with γ-Glu-Cys-Gly (glutathione), the dense taste of γ-Glu-Nva-Gly gives efficacy activities, and more the time of morning starts to strengthen after consumption.Thus, by sense evaluation test confirm γ-Glu-Nva-Gly for be not completely in taste type, but flavor strengthens a little earlier and lasts till that the beef extract of aftertaste has effect more more significant than γ-Glu-Cys-Gly (glutathione) and γ-Glu-Val-Gly.
Sense evaluation test is implemented as follows.Commercially available beef extract (solid constituent 61.2 % by weight, salinity 12.2 % by weight) is dissolved and is 3.0 % by weight in the hot water, be mixed with beef extract solution.For this beef extract solution, mix γ-Glu-Nva-Gly, the γ-Glu-Cys-Gly as sample or γ-Glu-Val-Gly.Measure and use paired comparison test's method, evaluation is compared: the γ-Glu-Nva-Gly of (1) 0.0003 % by weight gives the γ-Glu-Cys-Gly of 0.02 % by weight of activity with having equal dense taste to following, γ-the Glu-Val-Gly of the γ-Glu-Nva-Gly of (2) 0.0003 % by weight and 0.0003 % by weight of isodose, γ-the Glu-Nva-Gly of (3) 0.0003 % by weight gives the γ-Glu-Val-Gly of 0.002 % by weight of activity with having equal dense taste, and judge that whichever " strengthens its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change beef extract by judging panel, preferred ".Evaluate and implement with n=9.The judging panel's number γ-Glu-Nva-Gly of 0.0003 % by weight being evaluated as " strengthening its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change beef extract, is preferred " is as shown in table 10.
This result shows, even if under the condition that the equal dense taste such as shown in (1) and (3) is tired, γ-Glu-Nva-Gly also obviously " under the condition of the balance of the flavor and local flavor that do not change beef extract, strengthen its flavor and local flavor, be preferred ".
Table 10
N=9
*)in the level of signifiance of 1%, can think that γ-Glu-Nva-Gly strengthens its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change beef extract, be preferred
*)in the level of signifiance of 5%, can think that γ-Glu-Nva-Gly strengthens its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change beef extract, be preferred
Known with have compared with γ-Glu-Cys-Gly, γ-Glu-Val-Gly that equal dense taste gives under active concentration, the effect that γ-Glu-Nva-Gly has " strengthening its flavor and local flavor under the condition of the balance of the flavor and local flavor that do not change beef extract, is preferred " very gives prominence to like this.Beef source is worldwide widely used in flavoring, soup, poultry meat processed goods, the cooking processed goods, dessert, little food etc.
Therefore, γ-Glu-Nva-Gly can improve food with lower cost, small consumption, is also industrially very favorable.
Claims (6)
1. a food compositions, it contains
γ-the Glu-Nva-Gly of 0.01 ~ 3 weight ppm,
The pork source of 0.005 ~ 80 % by weight or beef source and
Other raw-food material.
2. food compositions according to claim 1, it uses pork source.
3. food compositions according to claim 1, it uses beef source.
4. the manufacture method of food compositions according to claim 1, it comprises the flavor reinforcing agent comprising γ-Glu-Nva-Gly to add and is mixed in pork source or beef source and other drink raw-food materials, make the γ-Glu-Nva-Gly concentration of diet product be the step of 0.01 ~ 3 weight ppm, and cook the step of the diet product raw mixture obtained as required further.
5. manufacture method according to claim 4, wherein, the described food compositions containing pork source is selected from pork extract, sausage and is noodle soup.
6. manufacture method according to claim 4, wherein, the described food compositions containing beef source is selected from beef extract, the Salted beef that salts down, use the soup of beef and use the sauce of beef.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2009297494 | 2009-12-28 | ||
| JP2009-297494 | 2009-12-28 | ||
| JP2010226571 | 2010-10-06 | ||
| JP2010-226571 | 2010-10-06 | ||
| PCT/JP2010/073722 WO2011081186A1 (en) | 2009-12-28 | 2010-12-28 | Flavor-enriching agent |
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| Publication Number | Publication Date |
|---|---|
| CN102753041A CN102753041A (en) | 2012-10-24 |
| CN102753041B true CN102753041B (en) | 2015-11-25 |
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| CN201080063730.6A Expired - Fee Related CN102753041B (en) | 2009-12-28 | 2010-12-28 | Dense taste imparting agent |
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| JP (1) | JP5850399B2 (en) |
| KR (1) | KR101512627B1 (en) |
| CN (1) | CN102753041B (en) |
| AU (1) | AU2010339306B2 (en) |
| CA (1) | CA2783415C (en) |
| IL (1) | IL220527A0 (en) |
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Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI606786B (en) | 2012-02-06 | 2017-12-01 | 味之素股份有限公司 | Composition for imparting rich taste to foods and beverages |
| JP2015097474A (en) * | 2012-03-09 | 2015-05-28 | 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 |
| WO2014123175A1 (en) * | 2013-02-07 | 2014-08-14 | 味の素株式会社 | Method for producing food/drink having augmented spice flavor |
| JP6557947B2 (en) * | 2014-07-11 | 2019-08-14 | 味の素株式会社 | W / O / W emulsion |
| JP6635268B2 (en) * | 2014-11-26 | 2020-01-22 | 不二製油株式会社 | Method for producing salty fat |
| JP6601578B2 (en) | 2017-07-13 | 2019-11-06 | 不二製油グループ本社株式会社 | peptide |
| CN107325156B (en) * | 2017-08-04 | 2020-09-08 | 北京工商大学 | Polypeptide with taste development property and polypeptide lipidization modification product |
| CN113316395A (en) | 2019-01-16 | 2021-08-27 | 不二制油集团控股株式会社 | Edible oil and fat composition and method for producing same |
| CN113543655A (en) * | 2019-03-05 | 2021-10-22 | 马斯公司 | Peptides that modulate calcium sensitive receptor activity to modulate kokumi taste and pet foods comprising the same |
| CN112931836A (en) * | 2021-03-23 | 2021-06-11 | 东北农业大学 | Novel flavor enhancer and preparation method thereof |
| KR20230090044A (en) * | 2021-12-14 | 2023-06-21 | 샘표식품 주식회사 | Manufacturing method for enhancement of gamma-glutamyl peptide for vegetable fermented products having deep taste |
Citations (1)
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|---|---|---|---|---|
| CN101305281A (en) * | 2005-11-09 | 2008-11-12 | 味之素株式会社 | KOKUMI-imparting agent |
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|---|---|---|---|---|
| RU2006138600A (en) * | 2004-04-06 | 2008-05-27 | Квест Интернэшнл Сервисиз Б.В. (Nl) | TASTING IMPROVING SUBSTANCES |
| EP2299271B9 (en) * | 2005-11-09 | 2014-02-26 | Ajinomoto Co., Inc. | Kokumi-imparting compositions |
| JPWO2007066430A1 (en) * | 2005-12-08 | 2009-05-14 | 協和発酵キリン株式会社 | Peptide production method |
| SG193061A1 (en) * | 2007-05-08 | 2013-09-30 | Ajinomoto Kk | Low-fat food |
| CN101677610B (en) * | 2007-05-08 | 2014-08-06 | 味之素株式会社 | Sweetener |
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- 2010-12-28 WO PCT/JP2010/073722 patent/WO2011081186A1/en active Application Filing
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101305281A (en) * | 2005-11-09 | 2008-11-12 | 味之素株式会社 | KOKUMI-imparting agent |
Also Published As
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| AU2010339306B2 (en) | 2013-06-13 |
| NZ601030A (en) | 2014-04-30 |
| AU2010339306A1 (en) | 2012-06-21 |
| CA2783415C (en) | 2015-11-24 |
| RU2532106C2 (en) | 2014-10-27 |
| CA2783415A1 (en) | 2011-07-07 |
| WO2011081186A1 (en) | 2011-07-07 |
| IL220527A0 (en) | 2012-08-30 |
| JP5850399B2 (en) | 2016-02-03 |
| KR101512627B1 (en) | 2015-04-16 |
| SG181978A1 (en) | 2012-08-30 |
| JPWO2011081186A1 (en) | 2013-05-13 |
| KR20120098946A (en) | 2012-09-05 |
| RU2012132448A (en) | 2014-02-10 |
| TW201136533A (en) | 2011-11-01 |
| CN102753041A (en) | 2012-10-24 |
| MX2012007244A (en) | 2012-07-30 |
| TWI565417B (en) | 2017-01-11 |
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