WO2011102310A1 - Blood uric acid level lowering agent - Google Patents
Blood uric acid level lowering agent Download PDFInfo
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- WO2011102310A1 WO2011102310A1 PCT/JP2011/053004 JP2011053004W WO2011102310A1 WO 2011102310 A1 WO2011102310 A1 WO 2011102310A1 JP 2011053004 W JP2011053004 W JP 2011053004W WO 2011102310 A1 WO2011102310 A1 WO 2011102310A1
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- cheese
- protease
- degradation product
- enzyme
- uric acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/20—Milk; Whey; Colostrum
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/06—Antigout agents, e.g. antihyperuricemic or uricosuric agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Definitions
- the present invention relates to a blood uric acid level lowering agent useful for the prevention and treatment of hyperuricemia.
- Hyperuricemia is a state in which the blood uric acid concentration (blood uric acid level) is high.
- the blood uric acid concentration abnormally increases, and inflammation that is caused by crystallization of those that could not be dissolved in the blood locally occurs in the body.
- gout attacks gout arthritis
- gout nodules when accumulated under the skin
- renal tubules and renal interstitium causess kidney damage.
- Non-Patent Document 1 reports that hyperuricemia itself is an independent risk factor for cardiovascular disorders have increased (see Non-Patent Document 1), and gout patients often have arteriosclerotic patients. This is a fact that has been well known since ancient times (for example, see Non-Patent Document 2). It has also been found that there is a very good correlation between the blood uric acid level and risk factors for metabolic syndrome (see Non-Patent Document 3).
- drugs that lower uric acid levels include drugs that promote excretion of uric acid, such as benzbromarone, and drugs that inhibit uric acid synthesis, such as allopurinol, and are prescribed according to the type of hyperuricemia in patients. Yes. These drugs currently prescribed are extremely excellent as a control agent for uric acid level, and the serum uric acid level can be returned to normal by taking the drug. However, since hyperuricemia itself is a very difficult disease to cure, if you stop taking the medicine, it will return to the state of hyperuricemia again.
- uric acid excretion enhancer increases the ability to excrete uric acid from the kidney by suppressing the physiological reabsorption of uric acid in the tubules, and lowers the serum uric acid level.
- side effects include gastrointestinal disorders, headache, and lightheadedness.
- severe liver damage may occur when administered to idiosyncratic patients (Non-patent Document 4).
- allopurinol As a uric acid production inhibitor, allopurinol has been introduced and widely used for the treatment of gout. This allopurinol inhibits xanthine oxidase, which acts at the final stage of the purine metabolic pathway, and reduces the amount of uric acid excreted in urine with a decrease in serum uric acid level.
- xanthine oxidase acts at the final stage of the purine metabolic pathway
- oxypurinol accumulates in the blood in a large amount and may cause a fatal poisoning syndrome (Non-patent Document 4).
- a therapeutic agent for hyperuricemia has been desired that can be safely and comfortably taken for a long period of time, for example, a product derived from a natural product that has been ingested daily since ancient times.
- chitosan Patent Document 1
- water-soluble dietary fiber Patent Document 2
- activated carbon Patent Document 3
- ⁇ -polylysine can be used as substances having the action of regulating the absorption of purines that cause hyperuricemia.
- cheese enzyme degradation products and peptides found from cheese enzyme degradation products include angiotensin converting enzyme inhibitory action (Patent Documents 5, 6, and 7), anti-inflammatory action (Patent Document 8), and gastrointestinal ulcer treatment effect ( Patent Document 9) is disclosed.
- Patent Documents 5, 6, and 7 angiotensin converting enzyme inhibitory action
- Patent Document 8 anti-inflammatory action
- Patent Document 9 gastrointestinal ulcer treatment effect
- JP 2001-163788 A Japanese Patent Laid-Open No. 2005-047828 JP 2005-187405 A JP 2006-001898 A WO2004 / 032642 WO2004 / 047543 WO2005 / 061529 WO2007 / 080962 JP 2009-120519 A
- An object of the present invention is to provide a blood uric acid level-lowering agent that is derived from a natural product and can be taken continuously for a long period of time.
- a cheese enzyme degradation product is It has been found that it has an excellent blood uric acid level lowering action and active oxygen absorption ability and is useful as a blood uric acid level lowering agent, and thus a prophylactic and therapeutic agent for hyperuricemia.
- the present invention (1) A blood uric acid level-lowering agent comprising a cheese enzyme degradation product as an active ingredient; (2) The blood uric acid level lowering agent according to (1), wherein the cheese enzyme degradation product is a cheese enzyme degradation product having active oxygen absorption ability; (3) The blood uric acid level-lowering agent according to (2), wherein the content of the enzyme degradation product of cheese is an amount showing an active oxygen absorption capacity of 50 ⁇ M Trolox equivalent or more as a daily dose; (4) The blood uric acid level-lowering agent according to any one of (1) to (3), wherein the cheese enzyme degradation product is a degradation product obtained by treating cheese with lipase and protease; (5) The blood uric acid level-lowering agent according to (4), wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme ; (6)
- lactis Lactococcus lactis subsp. Cremoris, and Lactococcus lactis subsp. the method according to (37), which is lactis biovar diacylactis; (39) The method according to (29), wherein substantially only a cheese enzyme degradation product is administered as an active ingredient; (40) The method of (39), wherein only the cheese enzyme degradation product and the formulation adjuvant are administered; (41) An active oxygen absorption method comprising administering a cheese enzyme degradation product; (42) The method according to (41), wherein substantially only a cheese enzyme degradation product is administered as an active ingredient, Is to provide. Moreover, this invention provides use of the cheese enzyme degradation product for blood uric acid value lowering agent or active enzyme absorber manufacture.
- the blood uric acid level-lowering agent of the present invention has an excellent blood uric acid level-lowering action, has a good flavor, and is highly safe. It is possible to control the blood uric acid level for a long period of time, and is useful for the prevention and treatment of hyperuricemia and gout.
- the active ingredient of the blood uric acid level-lowering agent and active oxygen absorber of the present invention is a cheese enzyme degradation product.
- the cheese enzyme degradation product is obtained by causing an enzyme selected from lipase and protease to act on cheese and degrading fat and protein in the cheese. More specifically, ripening is promoted by adding lipase, microorganisms such as acidic and neutral protease, and various animal-derived enzymes to green cheese, natural cheese after ripening or during ripening, and processed cheese.
- the cheese used as the raw material for the enzyme degradation product of cheese includes various cheeses including green cheese, and may be either natural cheese or processed cheese.
- the raw material of cheese is not particularly limited, and it may be made from human milk as well as milk from various mammals such as sheep and goats as well as cow's milk.
- the manufacturing method is not limited, and it is a cheese obtained using raw milk or powdered milk as a raw material, and various types such as fermented or ripened by various microorganisms such as lactic acid bacteria and mold as well as unripe cheese Cheese is available. For example, gouda cheese, cheddar cheese, skim cheese or the like can be used, but is not limited to this example.
- enzymes that decompose fats and proteins in cheese such as lipase and protease are used, but the type is not limited.
- lipase not only triacylglycerol lipase present in gastric juice but also various lipases isolated from human or mammalian body fluids, tissues thereof, insects, plants, bacteria, and the like are used.
- protease various proteases isolated from human or mammalian body fluids, tissues, insects, plants, bacteria, etc., such as pepsin, trypsin, and chymotrypsin secreted in the digestive tract are used.
- proteases There are two types of proteases, endo and exo, due to differences in the cleavage site of the amino acid sequence.
- the end type cleaves from the middle of the protein
- the exo type cleaves amino acids one by one from the C-terminal or N-terminal.
- the optimum pH, temperature, protein resolution, and amino acid cleavage site differ depending on the type of protease. For this reason, when treating with a protease, it is preferable to combine two or more, preferably three or more, proteases having different endo-type, exo-type, protein resolution, optimum pH and optimum temperature.
- a combination of two or more, particularly three or more selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme it is preferable to use a combination of two or more selected from endo-type proteases and protease / peptidase complex enzymes, particularly three or more.
- endo-type protease examples include trypsin (CAS No. 9002-07-7, EC 3.4.21.4, bovine pancreas-derived, Product No. T8802, SIGMA), pepsin (CAS No. 9001-75-6, EC 3.4.4.1, derived from porcine gastric mucosa, SIGMA), chymotrypsin (Novo, Boehringer), Proleather FG-F (from Bacillus subtilis; Amano Enzyme), biolase (from Bacillus subtilis, Nagase Sangyo), papain W-40 (Amano Enzyme), chymosin (from EC 3.4.23.4, Maxiren, modified yeast Kluyveromyces lactis, GIST-BROCADES NV), Alcala eR (Bacillus licheniformis derived from, Nobosha), Molsin F (Aspergillus saitoi origin, Kikkoman Corporation).
- trypsin CAS No. 9002-07-7
- Esperase from Bacillus lentus, Novo
- Neurase R from Bacillus subtilis, Novo
- Protamex from bacteria, Novo
- PTN6.0S pig pancreatic trypsin, Novo
- protease S "Amano” G from Bacillus stearothermophilus, Amano enzyme
- exo-type proteases examples include pancreatic carboxypeptidase and small intestinal brush border aminopeptidase.
- Examples of the endo-type / exo-type protease complex enzyme include neurolase A (derived from Aspergillus niger, Amano Enzyme), and examples of the exo-type peptidase / endo-type protease complex enzyme include flavorzyme (derived from Aspergillus oryzae, Novo). .
- protease / peptidase complex enzyme for example, protease A “Amano” SD (derived from Aspergillus oryzae, Amano Enzyme), equine enzyme G (peptidases and proteases, derived from Aspergillus oryzae, Amano Enzyme) can be used.
- protease A “Amano” SD derived from Aspergillus oryzae, Amano Enzyme
- equine enzyme G peptidases and proteases, derived from Aspergillus oryzae, Amano Enzyme
- two or more types selected from endo-type proteases and protease / peptidase complex enzymes it is preferable to use two or more types selected from endo-type proteases and protease / peptidase complex enzymes, and more than two types, particularly three or more types, may be used in combination of endo-type proteases and protease / peptidase complex enzymes. Is preferred. More specifically, it is particularly preferable to use a combination of two or more, particularly three or more selected from protease A “Amano” SD, morsin, neurase and equinezyme G.
- lipases examples include Sumiteam NLS (derived from Aspergillus niger, Shin Nippon Chemical Industry), Lipase A “Amano” 6 (derived from Aspergillus niger, Amano Enzyme), Lipase R “Amano” (derived from Penicillium roqueforti, Amano Enzyme “Renno Enzyme”) Trick esterases "(derived from cattle, manufactured by Lenco New Zealand) are mentioned, but pregastric esterases derived from cattle are particularly preferred.
- the enzyme treatment conditions are not particularly limited as long as the enzyme is not deactivated. Optimum conditions for the enzyme to be used are preferable from the viewpoint of promoting decomposition, but depending on the effect of the cheese to be processed, the enzyme to be used, and the target cheese enzyme degradation product, the pH and reaction temperature during the reaction, the enzyme and the substrate (Cheese) concentration and reaction time are adjusted. Moreover, when processing using 2 or more types of enzymes, you may process with all the enzymes at once, and can also process sequentially with each enzyme.
- the reaction temperature for the enzyme treatment in the present invention is usually 20 to 60 ° C., preferably 30 to 50 ° C.
- the reaction time for the enzyme treatment in the present invention is usually about 1 to 8 days, preferably about 2 to 4 days.
- the amount of enzyme added varies depending on the titer of the enzyme used, but is usually 0.01% or more, preferably 0.1 to 10% per substrate (cheese).
- Protease treatment can be accompanied by lactic acid bacteria fermentation.
- Lactococcus lactic acid bacteria can be preferably used.
- Specific examples of bacterial species belonging to Lactococcus lactic acid bacteria include Lactococcus garvieae, Lactococcus lactis, Lactococcus lactis subspice lactis subsp. Lactococcus lactis subsp. Lactococcus lactis subsp. Lactis biovar diacetylactis, Lactococcus pistium, Lactococcus plantarum and Lactococcus rafinolactol Not. In particular, Lactococcus lactis subsp.
- Lactic acid fermentation makes it possible to maintain the pH in the enzyme treatment step on the acidic side. Moreover, it may replace with lactic acid fermentation and may adjust pH using the organic acid normally used for a pharmaceutical and foodstuffs, such as a citric acid, gluconic acid, hydrochloric acid, acetic acid, lactic acid, phosphoric acid, and an inorganic acid.
- the pH should be 3.5 to 6.0, preferably 4.0 to 5.6.
- the cheese enzyme degradation product used in the present invention is particularly preferably a degradation product treated with lipase, protease and lactic acid bacteria.
- the reaction temperature for these treatments is usually 20 to 60 ° C., preferably 30 to 50 ° C.
- the reaction time for these treatments is generally about 1 to 8 days, preferably about 2 to 4 days.
- the amount of enzyme added varies depending on the titer of the enzyme used, but is usually 0.01% or more, preferably 0.1 to 10% per substrate (cheese).
- the obtained cheese enzyme degradation product has an excellent blood uric acid level lowering action as shown in the Examples described later, and is useful as a prophylactic / therapeutic agent for hyperuricemia and as a prophylactic / therapeutic agent for gout.
- the cheese enzyme decomposition product has an excellent active oxygen absorption ability and is useful as an active oxygen absorbent.
- the excellent blood uric acid level lowering action of the enzyme degradation product of cheese is considered to involve such active oxygen absorption ability, that is, antioxidant ability.
- the cheese enzyme degradation product can be used alone, but is often provided as various compositions together with a carrier that is substantially non-toxic.
- substantially non-toxic means that undesirable effects are not exerted even when ingested by humans or animals, and there is little risk of reducing the action of the cheese enzyme degradation product, which is pharmaceutically or hygienically acceptable.
- Various carriers are used. Examples of such carriers (hereinafter also referred to as formulation adjuvants) include various starches such as corn starch, lactose, crystalline cellulose, dextrose, mannitol, sucrose, sorbitol, gelatin, gum arabic, dicalcium phosphate, tricalcium phosphate.
- Excipients such as monocalcium phosphate, sodium phosphate, sodium carbonate, lubricants such as stearic acid, zinc stearate, calcium stearate or magnesium stearate, binders such as sucrose, polyethylene glycol, polyvinylpyrrolidone, benzoic PH adjusters such as acids, citric acid, tartaric acid, succinic acid, phosphoric acid, hydrochloric acid, and other suitable organic acids and inorganic acids or alkali metal (for example, sodium or potassium) salts, flavoring agents, flavoring agents, coloring agents, Disintegrant, solubilizer, suspension Agents, coating agents and the like.
- binders such as sucrose, polyethylene glycol, polyvinylpyrrolidone, benzoic PH adjusters such as acids, citric acid, tartaric acid, succinic acid, phosphoric acid, hydrochloric acid, and other suitable organic acids and inorganic acids or alkali metal (for example, sodium or potassium) salts,
- the cheese enzyme degradation product should be provided as a pharmaceutical composition to be administered orally, such as powders, granules, fine granules, dry syrups, tablets, capsules, liquids, drinks, etc. according to conventional methods. Can do. Further, other components such as an appropriate amount of vitamins, minerals, sugars, amino acids, peptides may be added. Further, the composition of the present invention is preferably a composition containing substantially only a cheese enzyme degradation product as an active ingredient, and more specifically, a composition containing only a cheese enzyme degradation product and a formulation adjuvant. It may be.
- the blood uric acid level-lowering agent of the present invention may be administered simultaneously or as a mixture with other active ingredients, for example, a composition having a blood uric acid level-lowering action different from the cheese enzyme degradation product.
- a composition having a blood uric acid level lowering action having a different action mechanism includes, for example, chitosan, water-soluble dietary fiber, activated carbon that adsorbs nucleic acid ingested as food and inhibits nucleic acid from being absorbed from the intestinal tract , ⁇ -polylysine, Atlantic nettle to increase urine volume and promote excretion of uric acid, celery seed extract, lactic acid bacteria to reduce serum uric acid level by degrading purine taken from diet and reducing absorption into the body Anserine, which increases the amount of purine recycle enzyme and suppresses the production of uric acid too much, can be mentioned.
- propolis having xanthine oxidase inhibitory activity Iris genus, cinnamon, cedron, Iwabenkei, Benikeiten, Galangal, Nutmeg, Hypericum perforatum, and Quercetin.
- the blood uric acid level-lowering agent of the present invention can be used not only in pharmaceuticals but also in the form of food and drink.
- prevention or treatment of hyperuricemia is expected by taking it as a special-purpose food such as a food for specified health use or a nutritional functional food.
- the form thereof is not limited, such as liquid, paste, solid, powder, and various foods (milk, processed milk, milk drink, soft drink, fermented milk, yogurt, cheese, bread, biscuits , Crackers, pizza crusts, ice creams, candy, formula milk, liquid foods, foods for the sick, foods for infants, foods such as infant formula, nutritional foods, dietary supplements, supplements, frozen foods, processed foods, etc. And may be ingested.
- the food and drink according to the present invention are mixed with water, protein, carbohydrates, lipids, vitamins, minerals, organic acids and inorganic acids, organic bases and inorganic bases, fruit juices, flavors, thickeners, etc. in cheese enzyme degradation products.
- the protein include whole milk powder, skim milk powder, partially skimmed milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, ⁇ -casein, ⁇ -casein, ⁇ -casein, ⁇ -lactoglobulin , ⁇ -lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and vegetable proteins and their degradation products.
- saccharide examples include saccharides such as sucrose and maltose, processed starch (in addition to dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like.
- lipids include animal oils such as lard and fish oil, fractionated oils, hydrogenated oil, transesterified oil, palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils.
- vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline.
- folic acid examples include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey mineral.
- the organic acid examples include malic acid, citric acid, lactic acid, tartaric acid, and examples of the inorganic acid include hydrochloric acid and phosphoric acid.
- the thickener include agar, gelatin, carrageenan, guar gum, xanthan gum and the like.
- milk-derived components such as butter, whey minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids and lactose, peptides such as casein phosphopeptides, and amino acids such as arginine and lysine. Also good.
- these components may be either synthetic products or products derived from natural products, or foods containing a large amount of these may be used as raw materials. These components can be used in combination of two or more.
- the blood uric acid level-lowering agent of the present invention is administered to a human or animal that exhibits hyperuricemia or a human or animal that is predicted to increase in uric acid level, and contains an effective amount of a cheese enzyme degradation product.
- the dosage is generally 0.01 mg / kg to 2000 mg / kg, preferably 1 mg / kg to 1000 mg / kg, more preferably 10 mg / kg to 500 mg / kg as a cheese enzyme degradation product per day. This dose is appropriately changed according to the age, weight, symptom and target site of the human or animal to be administered, and the type and amount of food to be ingested.
- the cheese enzyme degradation product has an active oxygen absorption action, and therefore, the dose can be limited based on the active oxygen absorption capacity (ORAC: Oxygen Radial Absorbance Capacity).
- Active oxygen absorption capacity is an index of antioxidant power developed by the US Department of Agriculture, National Institute for Aging. When a certain reactive oxygen species is generated in a system in which a fluorescent substance is present, the fluorescence intensity of the fluorescent substance decomposed thereby is measured, and when a fluorescence intensity curve that decreases with time is drawn, this reaction system is antioxidant When the substance coexists, the rate of decrease in the fluorescence intensity of the fluorescent substance is delayed.
- the difference (net AUC) between the area under the curve (AUC: Area Under the Curve) of the fluorescence intensity in the presence of the specimen (or standard substance) and the AUC in the absence (blank) is calculated.
- AUC Area Under the Curve
- a relative value with respect to the net AUC of a standard substance having a known concentration is determined. Based on the relative value, it is converted into the concentration of Trolox to obtain the antioxidant power of the specimen.
- the dose of the blood uric acid level-lowering agent of the present invention is an amount that exhibits an active oxygen absorption capacity of 50 ⁇ M Trolox equivalent or more, preferably 100 ⁇ M Trolox equivalent or more, more preferably 1000 ⁇ M Trolox equivalent or more as a daily dose. be able to.
- Example 1 (Preparation of starter culture) A 20 w skim milk powder was dissolved in 200 ml of distilled water and sterilized to prepare a 10 w / v% skim milk medium. This was inoculated with about 0.1 g of three starter bacteria (Lactococcus lactis subsp. Lactis, Lactococcus lactis subsp. Cremoris, Lactococcus lactis subsp. Lactis biovar diactylactis) at 37 ° C. for 16 hours.
- three starter bacteria Lactis subsp. Lactis, Lactococcus lactis subsp. Cremoris, Lactococcus lactis subsp. Lactis biovar diactylactis
- Protease A “Amano” SD Aspergillus oryzae-derived protease, Amano Enzyme
- lipase pregastric esterase, lotus root
- the pH was adjusted to 4.1 with citric acid
- Morsine F acidic protease derived from Aspergillus saitoi, manufactured by Kikkoman
- Umamizyme G protease derived from Aspergillus orizae, manufactured by Amano Enzyme
- Decomposition was performed by shaking at 0 ° C. Four days later, the pH was adjusted to 5.0 with sodium hydroxide, and the enzyme was deactivated by heating at 110 ° C. for 15 minutes to obtain a cheese enzyme decomposition product.
- the compounding amount per 100 g of domestic gouda cheese is as follows. [Composition table] 100g domestic gouda cheese Sterile water 50.5g Starter culture solution 18g Sodium chloride 2.2g Protease A “Amano” SD 0.6g Lipase 0.2g Citric acid 3.5g Morsin F 0.2g Ummamizyme G 0.3g Sodium hydroxide 1.5g
- Example 2 Preparation of cheese enzyme degradation product 2 After adding starter culture solution and sodium chloride prepared in Example 1 to domestic cheddar cheese added with sterilized water and stirring, protease A “Amano” SD (derived from Aspergillus oryzae, Amano Enzyme), lipase (pregastric) Esterase (manufactured by Lenco New Zealand) was added, and the mixture was shaken at 34 ° C. for decomposition. 48 hours later, the pH was adjusted to 4.1 with citric acid, and Neurase R (from Bacillus subtilis, Novo) and Umamizyme G (Aspergillus oryzae-derived protease, Amano Enzyme) were added, respectively. Decomposition was performed by shaking.
- protease A “Amano” SD derived from Aspergillus oryzae, Amano Enzyme
- lipase pregastric Esterase
- the blending amount per 100 g of domestic cheddar cheese is as follows. [Composition table] Domestic cheddar cheese 100g Sterile water 50.5g Starter culture solution 20g Sodium chloride 0.3g Protease A “Amano” SD 0.6g Lipase 0.2g Citric acid 3.3g Neurase R 0.2g Ummamizyme G 0.3g Sodium hydroxide 1.5g
- Example 3 Preparation of cheese enzyme degradation product 3) Danish skim cheese (ripening period 6 months) with distilled water added to the starter culture solution and sodium chloride prepared in Example 1, and after stirring, protease S “Amano” G (from Bacillus stearothermophilus, Amano Enzyme) Was added and shaken at 34 ° C. for decomposition. 48 hours later, the pH was adjusted to 4.1 with citric acid, and Neurase A (derived from Aspergillus niger, Amano Enzyme) and Umamizyme G (Aspergillus oryzae derived protease, Amano Enzyme) were added and shaken at 34 ° C. The decomposition was performed.
- Danish skim cheese 100g 74g of distilled water Starter culture solution 20g Sodium chloride 0.2g Protease S "Amano" G 0.6g Citric acid 3.2g Newase A 0.3g Ummamizyme G 0.3g Sodium hydroxide 1.4g
- Example 4 Effect of cheese enzyme degradation product on oxonate-induced hyperuricemia (animal experiment) Rats (Wistar, male, 7 weeks old, SLC Japan) were purchased and fed AIN-93G powder during the habituation period. After the acclimation period, blood was collected in the morning and the uric acid level was measured (day 0). Based on this value, it was divided into a total of 5 groups (6 mice per group): negative group, control group, test 1 group, test 2 group, and test 3 group.
- As the cheese enzyme degradation product one prepared according to the preparation of the cheese enzyme degradation product described in Example 1 was used.
- AIN-93G is a standard purified feed composition for nutritional research for mice and rats published in 1993 by the American Institute of Nutrition.
- Negative group AIN-93G feeding, water for injection (10 mL / kg)
- Control group Potassium oxonate 2.5% mixed AIN-93G feeding, water for injection (10 mL / kg)
- Test group 1 Feeding potassium oxonate 2.5% AIN-93G, cheese enzyme degradation product (300 mg / kg, 10 mL / kg)
- Test group 2 Feeding AIN-93G with potassium oxonate 2.5%, cheese enzyme degradation product (1 g / kg, 10 mL / kg)
- Test group 3 Feeding with 2.5% potassium oxonate mixed AIN-93G, enzyme degradation product of cheese (3 g / kg, 10 mL / kg)
- the measurement of drinking water is performed on the 0th day (set value), 2nd day (residual value, set value), 6th day (residual value, set value), 8th day (residual value, set value), 12th day ( Residual value, set value), 15th day (residual value, set value), 19th day (residual value, set value), and 22nd day (residual value).
- Example 5 Measurement of active oxygen absorption capacity of cheese enzymatic degradation product ORAC (Oxygen Radical Absorbance Capacity) was established in 1992 by the United States Department of Agriculture (USDA) and the National Institute on Aging. It is a new index of antioxidant power developed by researchers and is an excellent method for analyzing antioxidant power in foods. Currently, it has the most extensive database of food materials such as vegetables and fruits and processed foods. It is an analytical method.
- ORAC Oxygen Radical Absorbance Capacity
- the cheese enzyme degradation product used as a sample was prepared by adding an 8M urea solution to 30 mg / mL, and then the cheese enzyme degradation product was uniformly dispersed by ultrasonic treatment. A 10-fold dilution with 75 mM potassium phosphate buffer (pH 7.4) was performed twice. A labeling substance (Fluorescein: 3 ′, 6′-dihydroxyspiro [isobenzofuran-1 [3H], 9 ′ [9H] -xanthen] -3-one) is added with 75 mM potassium phosphate buffer (pH 7.4), and 117 nM Fluorescein Was prepared.
- 75 mM potassium phosphate buffer (pH 7.4) was added to a standard substance (Trolox TM : 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) to prepare 50 ⁇ M Trolox.
- a radical generator (AAPH: 2,2′-azo-bis (2-amidinopropane) dihydrochloride) was added with 75 mM potassium phosphate buffer (pH 7.4) to prepare 40 mM AAPH.
- 20 ⁇ l of sample (or 50 ⁇ M Trolox of standard substance) and 120 ⁇ l of 117 nM Fluorescein were added and pre-incubated at 37 ° C. for 15 minutes. Thereafter, 60 ⁇ l of 40 mM AAPH as a radical generator was added, and the fluorescence intensity was measured every minute at an excitation wavelength of 485 nm and a fluorescence wavelength of 520 nm.
- the active oxygen absorption capacity of the cheese enzymatic degradation product was 189.5 ⁇ 11.0 ⁇ M Trolox equivalent / gram.
- Example 6 Production of fermented milk (Production Example 1 of fermented milk) Plain yogurt was dissolved in a cheese enzyme digest (Meiji cheese taste concentrate G (Gouda type)), L. bulgaricus JCM1002 T , S. prepared by thermophilus ATCC 19258. First, using a skim milk powder 10% medium, L. bulgaricus JCM1002 T , S. A bulk starter of thermophilus ATCC 19258 was prepared. Next, the cheese enzyme degradation product was mixed with yoghurt mix (non-fat milk solids (SNF): 9.5%, fat (FAT): 3.0%), and heat-treated at 95 ° C for 5 minutes. In the mix after this heat treatment, L.
- SNF non-fat milk solids
- FAT fat
- bulgaricus JCM1002T Thermophilus ATCC 19258 starter was inoculated at 1% each and fermented at 43 ° C. for 4 hours to obtain plain yogurt.
- the plain yogurt was cooled in a refrigerator (5 ° C.), and then the flavor and physical properties were confirmed. At this time, both flavor and physical properties were good.
- the cheese enzyme degradation product was mixed with yoghurt mix (non-fat milk solids (SNF): 9.5%, fat (FAT): 3.0%), and heat-treated at 95 ° C for 5 minutes.
- SNF non-fat milk solids
- FAT fat
- L. bulgaricus JCM1002 T thermophilus ATCC 19258 starter 1% each
- L.P. Gasseri OLL2959 starter was inoculated at 5% and fermented at 43 ° C. for 4 hours to obtain plain yogurt.
- the plain yogurt was cooled in a refrigerator (5 ° C.), and then the flavor and physical properties were confirmed. At this time, both flavor and physical properties were good.
- Example 8 Production of process cheese (production example of process cheese) New Zealand cheddar cheese was used as a raw material cheese, and an enzyme digestion product having an active oxygen absorption capacity of about 150 ⁇ M Trolox equivalent / g was added to prepare a processed cheese by the following production method.
- the blending ratio is as follows.
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Abstract
Description
(1)チーズ酵素分解物を有効成分とする血中尿酸値低下剤;
(2)チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、(1)記載の血中尿酸値低下剤;
(3)チーズ酵素分解物の含有量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、(2)記載の血中尿酸値低下剤;
(4)チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、(1)~(3)のいずれか1項記載の血中尿酸値低下剤;
(5)プロテアーゼが、エンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(4)記載の血中尿酸値低下剤;
(6)プロテアーゼが、エンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(4)記載の血中尿酸値低下剤;
(7)エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、請求項6記載の血中尿酸値低下剤;
(8)リパーゼが、ウシ由来のプレガストリックエステラーゼである、(4)記載の血中尿酸値低下剤;
(9)チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、(1)~(3)のいずれか1項記載の血中尿酸値低下剤;
(10)乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、(9)記載の血中尿酸値低下剤;
(11)有効成分としてチーズ酵素分解物のみを含有する、(1)~(10)のいずれか1つに記載の血中尿酸値低下剤;
(12)チーズ酵素分解物および製剤補助剤のみを含有する、請求項1~11のいずれか1つに記載の血中尿酸値低下剤;
(13)チーズ酵素分解物を有効成分とする活性酸素吸収剤;
(14)有効成分としてチーズ酵素分解物のみを含有する、(13)記載の活性酸素吸収剤;
(15)血中尿酸値を低下させるためのチーズ酵素分解物を含有する組成物;
(16)チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、(15)記載の組成物;
(17)チーズ酵素分解物の含有量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、(16)記載の組成物;
(18)チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、請求項(15)~(17)のいずれか1つに記載の組成物;
(19)プロテアーゼがエンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(18)記載の組成物;
(20)プロテアーゼがエンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(18)記載の組成物;
(21)エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、(20)記載の組成物;
(22)リパーゼがウシ由来のプレガストリックエステラーゼである、(18)記載の組成物;
(23)チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、(15)~(17)のいずれか1つに記載の組成物;
(24)乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、(23)記載の組成物;
(25)有効成分としてチーズ酵素分解物のみを含有する、(15)~(24)のいずれか1つに記載の組成物;
(26)チーズ酵素分解物および製剤補助剤のみを含有する、請求項15~25のいずれか1つに記載の組成物;
(27)活性酵素を吸収するためのチーズ酵素分解物を含有する組成物;
(28)有効成分としてチーズ酵素分解物のみを含有する、(27)記載の組成物;
(29)チーズ酵素分解物を投与することを特徴とする血中尿酸値の低下方法;
(30)チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、(29)記載の方法;
(31)チーズ酵素分解物の投与量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、(30)記載の方法;
(32)チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、(29)記載の方法;
(33)プロテアーゼがエンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(32)記載の方法;
(34)プロテアーゼがエンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、(32)記載の方法;
(35)エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、(34)記載の方法;
(36)リパーゼがウシ由来のプレガストリックエステラーゼである、(32)記載の方法;
(37)チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、(29)記載の方法;
(38)乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、(37)記載の方法;
(39)有効成分として、実質的にチーズ酵素分解物のみを投与する、(29)記載の方法;
(40)チーズ酵素分解物および製剤補助剤のみを投与する、(39)の方法;
(41)チーズ酵素分解物を投与することを特徴とする活性酸素吸収方法;
(42)有効成分として、実質的にチーズ酵素分解物のみを投与する、(41)記載の方法、
を提供するものである。
また、本発明は、血中尿酸値低下剤又は活性酵素吸収剤製造のためのチーズ酵素分解物の使用を提供するものである。 That is, the present invention
(1) A blood uric acid level-lowering agent comprising a cheese enzyme degradation product as an active ingredient;
(2) The blood uric acid level lowering agent according to (1), wherein the cheese enzyme degradation product is a cheese enzyme degradation product having active oxygen absorption ability;
(3) The blood uric acid level-lowering agent according to (2), wherein the content of the enzyme degradation product of cheese is an amount showing an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose;
(4) The blood uric acid level-lowering agent according to any one of (1) to (3), wherein the cheese enzyme degradation product is a degradation product obtained by treating cheese with lipase and protease;
(5) The blood uric acid level-lowering agent according to (4), wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme ;
(6) The blood uric acid level lowering agent according to (4), wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme;
(7) The blood uric acid level-lowering agent according to claim 6, wherein the endo-type protease is an acidic protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae;
(8) The blood uric acid level-lowering agent according to (4), wherein the lipase is bovine-derived pregastric esterase;
(9) The agent for lowering blood uric acid level according to any one of (1) to (3), wherein the cheese enzyme degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria;
(10) Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. Cremoris, and Lactococcus lactis subsp. the blood uric acid level-lowering agent according to (9), which is lactis biovar diacylactis;
(11) The blood uric acid level lowering agent according to any one of (1) to (10), which contains only a cheese enzyme degradation product as an active ingredient;
(12) The blood uric acid level-lowering agent according to any one of
(13) an active oxygen absorbent comprising a cheese enzyme degradation product as an active ingredient;
(14) The active oxygen absorbent according to (13), which contains only a cheese enzyme degradation product as an active ingredient;
(15) A composition containing a cheese enzyme degradation product for lowering blood uric acid levels;
(16) The composition according to (15), wherein the cheese enzymatic decomposition product is a cheese enzymatic decomposition product having active oxygen absorption ability;
(17) The composition according to (16), wherein the content of the cheese enzyme degradation product is an amount showing an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose;
(18) The composition according to any one of (15) to (17), wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with a lipase and a protease;
(19) The composition according to (18), wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme;
(20) The composition according to (18), wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme;
(21) The composition according to (20), wherein the endo-type protease is an acidic protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae;
(22) The composition according to (18), wherein the lipase is a bovine-derived pregastric esterase;
(23) The composition according to any one of (15) to (17), wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria;
(24) Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. Cremoris, and Lactococcus lactis subsp. the composition according to (23), which is lactis biovar diacylactis;
(25) The composition according to any one of (15) to (24), which contains only a cheese enzyme degradation product as an active ingredient;
(26) The composition according to any one of
(27) A composition containing a cheese enzyme degradation product for absorbing an active enzyme;
(28) The composition according to (27), which contains only cheese degradation products as an active ingredient;
(29) A method for lowering blood uric acid level, which comprises administering a cheese enzymatic degradation product;
(30) The method according to (29), wherein the cheese enzymatic degradation product is a cheese enzymatic degradation product having active oxygen absorption ability;
(31) The method according to (30), wherein the dose of the cheese enzyme degradation product is an amount showing an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose;
(32) The method according to (29), wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with a lipase and a protease;
(33) The method according to (32), wherein the protease is at least two selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme;
(34) The method according to (32), wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme;
(35) The method according to (34), wherein the endo-type protease is an acidic protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae;
(36) The method according to (32), wherein the lipase is a bovine-derived pregastric esterase;
(37) The method according to (29), wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria;
(38) Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. Cremoris, and Lactococcus lactis subsp. the method according to (37), which is lactis biovar diacylactis;
(39) The method according to (29), wherein substantially only a cheese enzyme degradation product is administered as an active ingredient;
(40) The method of (39), wherein only the cheese enzyme degradation product and the formulation adjuvant are administered;
(41) An active oxygen absorption method comprising administering a cheese enzyme degradation product;
(42) The method according to (41), wherein substantially only a cheese enzyme degradation product is administered as an active ingredient,
Is to provide.
Moreover, this invention provides use of the cheese enzyme degradation product for blood uric acid value lowering agent or active enzyme absorber manufacture.
このうち、エンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上、特に3種以上を組み合せて用いるのが好ましい。さらに、エンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上、特に3種以上を組み合せて用いるのが好ましい。 There are two types of proteases, endo and exo, due to differences in the cleavage site of the amino acid sequence. The end type cleaves from the middle of the protein, and the exo type cleaves amino acids one by one from the C-terminal or N-terminal. The optimum pH, temperature, protein resolution, and amino acid cleavage site differ depending on the type of protease. For this reason, when treating with a protease, it is preferable to combine two or more, preferably three or more, proteases having different endo-type, exo-type, protein resolution, optimum pH and optimum temperature.
Among these, it is preferable to use a combination of two or more, particularly three or more selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme. Furthermore, it is preferable to use a combination of two or more selected from endo-type proteases and protease / peptidase complex enzymes, particularly three or more.
活性酸素吸収能力は、米国農務省、国立老化研究所で開発された抗酸化力の指標である。蛍光物質の存在する系において一定の活性酸素種を発生させ、それによって分解される蛍光物質の蛍光強度を測定し、経時的に減少する蛍光強度の曲線を描いた場合、この反応系に抗酸化物質が共存すると蛍光物質の蛍光強度の減少速度が遅延する。この原理に基づき、検体(もしくは標準物質)存在下での蛍光強度の曲線下面積(AUC:Area Under the Curve)と、非存在下(ブランク)でのAUCとの差(net AUC)を算出する。検体のnet AUCについて、濃度既知の標準物質(TroloxTM:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid:Troloxは登録商標である)のnet AUCに対する相対値を求める。その相対値を基にTroloxの濃度に換算して検体の抗酸化力とする。(単位μmol TE/g:マイクロモルTrolox当量/グラム)
本発明の血中尿酸値低下剤の投与量は、1日投与量として50μM Trolox当量以上、好ましくは100μM Trolox等量以上、より好ましくは1000μM Trolox等量以上の活性酸素吸収能を示す量とすることができる。 In addition, as described above, the cheese enzyme degradation product has an active oxygen absorption action, and therefore, the dose can be limited based on the active oxygen absorption capacity (ORAC: Oxygen Radial Absorbance Capacity).
Active oxygen absorption capacity is an index of antioxidant power developed by the US Department of Agriculture, National Institute for Aging. When a certain reactive oxygen species is generated in a system in which a fluorescent substance is present, the fluorescence intensity of the fluorescent substance decomposed thereby is measured, and when a fluorescence intensity curve that decreases with time is drawn, this reaction system is antioxidant When the substance coexists, the rate of decrease in the fluorescence intensity of the fluorescent substance is delayed. Based on this principle, the difference (net AUC) between the area under the curve (AUC: Area Under the Curve) of the fluorescence intensity in the presence of the specimen (or standard substance) and the AUC in the absence (blank) is calculated. . With respect to the net AUC of the specimen, a relative value with respect to the net AUC of a standard substance having a known concentration (Trolox ™ : 6-hydroxy-2,5,7,8-tetramethyl-2-carbon acid: Trolox is a registered trademark) is determined. Based on the relative value, it is converted into the concentration of Trolox to obtain the antioxidant power of the specimen. (Unit: μmol TE / g: micromol Trolox equivalent / gram)
The dose of the blood uric acid level-lowering agent of the present invention is an amount that exhibits an active oxygen absorption capacity of 50 μM Trolox equivalent or more, preferably 100 μM Trolox equivalent or more, more preferably 1000 μM Trolox equivalent or more as a daily dose. be able to.
(スターター培養液の調製)
20gのスキムミルク粉末に200mlの蒸留水に溶解し、滅菌して、10w/v%脱脂粉乳培地を調製した。ここに約0.1gのスターター菌3種(Lactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、Lactococcus lactis subsp.lactis biovar diacetylactis)を接種し、37℃で16時間培養した。
(チーズ酵素分解物の調製1)
国産ゴーダチーズに滅菌水を加えたものに、スターター培養液、塩化ナトリウムを添加し撹拌後、プロテアーゼA「アマノ」SD(Aspergillus oryzae由来プロテアーゼ、天野エンザイム社製)、リパーゼ(プレガストリックエステラーゼ、レンコニュージーランド社製)を添加し、34℃で振とうすることにより分解を行った。48時間後、クエン酸にてpHを4.1に調整し、モルシンF(Aspergillus saitoi由来酸性プロテアーゼ、キッコーマン社製)及びウマミザイムG(Aspergillus orizae由来プロテアーゼ、天野エンザイム社製)をそれぞれ添加し、34℃で振とうすることにより分解を行った。4日後、水酸化ナトリウムでpHを5.0に調整し、110℃にて15分間加熱して酵素を失活させ、チーズ酵素分解物を得た。国産ゴーダチーズ100gあたりの配合量は次の通りである。
〔配合表〕
国産ゴーダチーズ 100g
滅菌水 50.5g
スターター培養液 18g
塩化ナトリウム 2.2g
プロテアーゼA「アマノ」SD 0.6g
リパーゼ 0.2g
クエン酸 3.5g
モルシンF 0.2g
ウマミザイムG 0.3g
水酸化ナトリウム 1.5g [Example 1]
(Preparation of starter culture)
A 20 w skim milk powder was dissolved in 200 ml of distilled water and sterilized to prepare a 10 w / v% skim milk medium. This was inoculated with about 0.1 g of three starter bacteria (Lactococcus lactis subsp. Lactis, Lactococcus lactis subsp. Cremoris, Lactococcus lactis subsp. Lactis biovar diactylactis) at 37 ° C. for 16 hours.
(Preparation of cheese enzyme degradation product 1)
Protease A “Amano” SD (Aspergillus oryzae-derived protease, Amano Enzyme), lipase (pregastric esterase, lotus root) after stirring and adding starter culture solution and sodium chloride to domestic gouda cheese with sterilized water New Zealand) was added and the mixture was shaken at 34 ° C. for decomposition. 48 hours later, the pH was adjusted to 4.1 with citric acid, and Morsine F (acidic protease derived from Aspergillus saitoi, manufactured by Kikkoman) and Umamizyme G (protease derived from Aspergillus orizae, manufactured by Amano Enzyme) were added, respectively. Decomposition was performed by shaking at 0 ° C. Four days later, the pH was adjusted to 5.0 with sodium hydroxide, and the enzyme was deactivated by heating at 110 ° C. for 15 minutes to obtain a cheese enzyme decomposition product. The compounding amount per 100 g of domestic gouda cheese is as follows.
[Composition table]
100g domestic gouda cheese
Sterile water 50.5g
Starter culture solution 18g
Sodium chloride 2.2g
Protease A “Amano” SD 0.6g
Lipase 0.2g
Citric acid 3.5g
Morsin F 0.2g
Ummamizyme G 0.3g
Sodium hydroxide 1.5g
(チーズ酵素分解物の調製2)
国産チェダーチーズに滅菌水を加えたものに、実施例1で調製したスターター培養液、塩化ナトリウムを添加し撹拌後、プロテアーゼA「アマノ」SD(Aspergillus orizae由来、天野エンザイム)、リパーゼ(プレガストリックエステラーゼ、レンコニュージーランド社製)を添加し、34℃で振とうすることにより分解を行った。48時間後、クエン酸にてpHを4.1に調整し、ニューラーゼR(Bacillus subtilis由来、ノボ社)及びウマミザイムG(Aspergillus orizae由来プロテアーゼ、天野エンザイム社製)をそれぞれ添加し、34℃で振とうすることにより分解を行った。4日後、水酸化ナトリウムでpHを5.0に調整し、110℃にて15分間加熱して酵素を失活させ、チーズ酵素分解物を得た。国産チェダーチーズ100gあたりの配合量は次の通りである。
〔配合表〕
国産チェダーチーズ 100g
滅菌水 50.5g
スターター培養液 20g
塩化ナトリウム 0.3g
プロテアーゼA「アマノ」SD 0.6g
リパーゼ 0.2g
クエン酸 3.3g
ニューラーゼR 0.2g
ウマミザイムG 0.3g
水酸化ナトリウム 1.5g [Example 2]
(Preparation of cheese enzyme degradation product 2)
After adding starter culture solution and sodium chloride prepared in Example 1 to domestic cheddar cheese added with sterilized water and stirring, protease A “Amano” SD (derived from Aspergillus oryzae, Amano Enzyme), lipase (pregastric) Esterase (manufactured by Lenco New Zealand) was added, and the mixture was shaken at 34 ° C. for decomposition. 48 hours later, the pH was adjusted to 4.1 with citric acid, and Neurase R (from Bacillus subtilis, Novo) and Umamizyme G (Aspergillus oryzae-derived protease, Amano Enzyme) were added, respectively. Decomposition was performed by shaking. Four days later, the pH was adjusted to 5.0 with sodium hydroxide, and the enzyme was deactivated by heating at 110 ° C. for 15 minutes to obtain a cheese enzyme decomposition product. The blending amount per 100 g of domestic cheddar cheese is as follows.
[Composition table]
Domestic cheddar cheese 100g
Sterile water 50.5g
Starter culture solution 20g
Sodium chloride 0.3g
Protease A “Amano” SD 0.6g
Lipase 0.2g
Citric acid 3.3g
Neurase R 0.2g
Ummamizyme G 0.3g
Sodium hydroxide 1.5g
(チーズ酵素分解物の調製3)
デンマークスキムチーズ(熟成期間6ヶ月)に蒸留水を加えたものに、実施例1で調製したスターター培養液、塩化ナトリウムを添加し撹拌後、プロテアーゼS「アマノ」G(Bacillus stearothermophilus由来、天野エンザイム)を添加し、34℃で振とうすることにより分解を行った。48時間後、クエン酸にてpHを4.1に調整し、ニューラーゼA(Aspergillus niger由来、天野エンザイム)及びウマミザイムG(Aspergillus orizae由来プロテアーゼ、天野エンザイム)をそれぞれ添加し、34℃で振とうすることにより分解を行った。5日後、水酸化ナトリウムでpHを5.0に調整し、110℃にて15分間加熱して酵素を失活させ、チーズ酵素分解物を得た。デンマークスキムチーズ100gあたりの配合量は次の通りである。
〔配合表〕
デンマークスキムチーズ 100g
蒸留水 74g
スターター培養液 20g
塩化ナトリウム 0.2g
プロテアーゼS「アマノ」G 0.6g
クエン酸 3.2g
ニューラーゼA 0.3g
ウマミザイムG 0.3g
水酸化ナトリウム 1.4g Example 3
(Preparation of cheese enzyme degradation product 3)
Danish skim cheese (ripening period 6 months) with distilled water added to the starter culture solution and sodium chloride prepared in Example 1, and after stirring, protease S “Amano” G (from Bacillus stearothermophilus, Amano Enzyme) Was added and shaken at 34 ° C. for decomposition. 48 hours later, the pH was adjusted to 4.1 with citric acid, and Neurase A (derived from Aspergillus niger, Amano Enzyme) and Umamizyme G (Aspergillus oryzae derived protease, Amano Enzyme) were added and shaken at 34 ° C. The decomposition was performed. Five days later, the pH was adjusted to 5.0 with sodium hydroxide, and the enzyme was deactivated by heating at 110 ° C. for 15 minutes to obtain a cheese enzyme degradation product. The blending amount per 100 g of Danish skim cheese is as follows.
[Composition table]
Danish skim cheese 100g
74g of distilled water
Starter culture solution 20g
Sodium chloride 0.2g
Protease S "Amano" G 0.6g
Citric acid 3.2g
Newase A 0.3g
Ummamizyme G 0.3g
Sodium hydroxide 1.4g
(動物実験)
ラット(Wistar、雄、7週令、日本エスエルシー)を購入し、馴化期間中は、AIN-93G粉末を給餌した。馴化期間終了後、午前中に採血し、尿酸値を測定した(0日目)。この値を基に、陰性群、対照群、被験1群、被験2群、被験3群の計5群(1群6匹)に群分けした。チーズ酵素分解物は実施例1に記載のチーズ酵素分解物の調製に従って作成したものを用いた。
なお、AIN-93Gは、米国国立栄養研究所(American Institute of Nutrition)から1993年に発表されたマウス・ラット用の栄養研究のための標準精製飼料組成である。
・陰性群:AIN-93G給餌、注射用水(10mL/kg)
・対照群:オキソニン酸カリウム2.5%混合AIN-93G給餌、注射用水(10mL/kg)
・被験1群:オキソニン酸カリウム2.5%混合AIN-93G給餌、チーズ酵素分解物(300mg/kg、10mL/kg)
・被験2群:オキソニン酸カリウム2.5%混合AIN-93G給餌、チーズ酵素分解物(1g/kg、10mL/kg)
・被験3群:オキソニン酸カリウム2.5%混合AIN-93G給餌、チーズ酵素分解物(3g/kg、10mL/kg) [Example 4] Effect of cheese enzyme degradation product on oxonate-induced hyperuricemia (animal experiment)
Rats (Wistar, male, 7 weeks old, SLC Japan) were purchased and fed AIN-93G powder during the habituation period. After the acclimation period, blood was collected in the morning and the uric acid level was measured (day 0). Based on this value, it was divided into a total of 5 groups (6 mice per group): negative group, control group,
AIN-93G is a standard purified feed composition for nutritional research for mice and rats published in 1993 by the American Institute of Nutrition.
Negative group: AIN-93G feeding, water for injection (10 mL / kg)
Control group: Potassium oxonate 2.5% mixed AIN-93G feeding, water for injection (10 mL / kg)
Test group 1: Feeding potassium oxonate 2.5% AIN-93G, cheese enzyme degradation product (300 mg / kg, 10 mL / kg)
Test group 2: Feeding AIN-93G with potassium oxonate 2.5%, cheese enzyme degradation product (1 g / kg, 10 mL / kg)
Test group 3: Feeding with 2.5% potassium oxonate mixed AIN-93G, enzyme degradation product of cheese (3 g / kg, 10 mL / kg)
群分け後より、陰性群にはAIN-93Gを引き続き給餌し、その他の群にはオキソニン酸カリウム2.5%混合AIN-93G(試験食)を給餌した。チーズ酵素分解物投与は1日目から15日目の毎日、午前中に1回行った。部分採血は、全例において2日目(投与2時間後)、5日目(投与2時間後)、8日目(投与2時間後)、12日目(投与2時間後)、15日目(投与2時間後)に行い、血清尿酸値をリンタングステン酸法で測定した。15日目のチーズ酵素分解物投与以降から22日目までは後観察期間とした。22日目の午前中に採血し、血清尿酸値をリンタングステン酸法で測定した。 [Main breeding (examination)]
After grouping, the negative group continued to be fed with AIN-93G, and the other groups were fed with 2.5% potassium oxonate mixed AIN-93G (test meal). Administration of the enzyme degradation product of cheese was performed once in the morning every day from
・一般状態の観察と体重の測定
体重測定は、0日目、1日目、5日目、8日目、12日目、15日目及び22日目に行った。
・摂餌量と摂水量の測定
摂餌量の測定は0日目(セット値)、5日目(残値、セット値)、8日目(残値、セット値)、12日目(残値、セット値)、15日目(残値、セット値)、19日目(残値、セット値)、22日目(残値)に行った。飲水量の測定は0日目(セット値)、2日目(残値、セット値)、6日目(残値、セット値)、8日目(残値、セット値)、12日目(残値、セット値)、15日目(残値、セット値)、19日目(残値、セット値)、22日目(残値)に行った。 [Measurement and inspection, etc.]
-Observation of general condition and measurement of body weight Body weight measurement was performed on the 0th day, 1st day, 5th day, 8th day, 12th day, 15th day and 22nd day.
・ Measurement of food intake and water intake Measurement of food intake is on the 0th day (set value), 5th day (remaining value, set value), 8th day (remaining value, set value), 12th day (remaining value) Value, set value), 15th day (residual value, set value), 19th day (residual value, set value), and 22nd day (residual value). The measurement of drinking water is performed on the 0th day (set value), 2nd day (residual value, set value), 6th day (residual value, set value), 8th day (residual value, set value), 12th day ( Residual value, set value), 15th day (residual value, set value), 19th day (residual value, set value), and 22nd day (residual value).
結果は平均値±標準偏差で示し、対照群と菌体投与各群を比較した。数値化した検査値の分散比はF検定を行い、等分散の場合にはStudent’s t-検定、Dunnett 検定又はTukey-Kramer検定を、不等分散の場合にはMann-Whitney U検定を行った。 [Statistical processing]
The results are shown as mean ± standard deviation, and the control group and each group administered with the fungus body were compared. F-test is performed for the variance ratio of the numerical test values. The Student's t-test, Dunnett test or Tukey-Kramer test is performed for equal variances, and the Mann-Whitney U test is performed for unequal variances. It was.
体重測定の結果を表1に、摂餌量及び摂水量の結果を表2に、血清尿酸値の推移を図1及び表3に示す。 〔result〕
The results of body weight measurement are shown in Table 1, the results of food intake and water intake are shown in Table 2, and the transition of serum uric acid level is shown in FIG. 1 and Table 3.
オキソニン酸カリウム混合飼料による飼育(対照群、被験1群、被験2群、被験3群)は、AIN-93Gによる飼育(陰性群)に比較して、摂餌量が減少し、飲水量が増加した。試験物質を投与した被験1群、被験2群、被験3群について、対照群と比較して有意な摂餌量の変化、飲水量の変化は認められなかった(Dunnett test)。
オキソニン酸カリウム混合飼料による飼育により、尿酸値が有意に上昇し高尿酸血症状態となった(陰性群vs対照群、p<0.001,Student’s t-test)。チーズ酵素分解物の投与は、尿酸値の上昇を用量依存的に抑制した。対照群と比較して、チーズ酵素分解物1000mg/kgの投与(被験2群)では2日目及び12日目において、チーズ酵素分解物3000mg/kgの投与(被験3群)では2,5,8,12,15日目において有意に尿酸値が低下していた(Dunnett test,p<0.05)。なお、試験物質の投与を中止してから7日後(22日目)には、対照群、被験1群、被験2群、被験3群の間に有意な尿酸値の差は見られなくなった。 Throughout the study period, each group gained weight. There was no significant difference between groups (Tukey-Kramer test).
Feeding with potassium oxonate mixed feed (control group,
Breeding with the potassium oxonate mixed feed resulted in a significant increase in uric acid levels and hyperuricemia (negative group vs control group, p <0.001, Student's t-test). Administration of the cheese enzymatic degradation product suppressed the increase in uric acid levels in a dose-dependent manner. Compared with the control group, cheese enzyme degradation product 1000 mg / kg administration (test group 2) on
活性酸素吸収能力(ORAC:Oxygen Radical Absorbance Capacity)は1992年に米国農務省(USDA)と国立老化研究所(National Institute on Aging)の研究者らにより開発された抗酸化力の新しい指標で、食品中の抗酸化力を分析する方法として優れており、現在、野菜・果物などの食品素材や加工食品に至るまで最もデータベースが充実した分析法である。 [Example 5] Measurement of active oxygen absorption capacity of cheese enzymatic degradation product ORAC (Oxygen Radical Absorbance Capacity) was established in 1992 by the United States Department of Agriculture (USDA) and the National Institute on Aging. It is a new index of antioxidant power developed by researchers and is an excellent method for analyzing antioxidant power in foods. Currently, it has the most extensive database of food materials such as vegetables and fruits and processed foods. It is an analytical method.
サンプルとなるチーズ酵素分解物は、8M尿素溶液を添加し30mg/mLに調製後、超音波処理にてチーズ酵素分解物を均一に分散させた。75mMリン酸カリウム緩衝液(pH7.4)で10倍希釈を2回行った。
標識物質(Fluorescein:3’,6’-dihydroxyspiro[isobenzofuran-1[3H],9’[9H]-xanthen]-3-one)に75mMリン酸カリウム緩衝液(pH7.4)を添加し117nM Fluoresceinを調製した。標準物質(TroloxTM:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid)に75mMリン酸カリウム緩衝液(pH7.4)を添加し50μM Troloxを調製した。ラジカル発生剤(AAPH:2,2’-azo-bis(2-amidinopropane)dihydrochloride)に75mMリン酸カリウム緩衝液(pH7.4)を添加し40mM AAPHに調製した。96穴蛍光プレートに、サンプル(又は標準物質の50μM Trolox)を20μl、117nM Fluoresceinを120μl添加し、37℃で15分プレインキュベートした。その後、ラジカル発生剤の40mM AAPHを60μl添加し、励起波長485nm、蛍光波長520nmで1分ごとに蛍光強度を測定した。 [Measurement]
The cheese enzyme degradation product used as a sample was prepared by adding an 8M urea solution to 30 mg / mL, and then the cheese enzyme degradation product was uniformly dispersed by ultrasonic treatment. A 10-fold dilution with 75 mM potassium phosphate buffer (pH 7.4) was performed twice.
A labeling substance (Fluorescein: 3 ′, 6′-dihydroxyspiro [isobenzofuran-1 [3H], 9 ′ [9H] -xanthen] -3-one) is added with 75 mM potassium phosphate buffer (pH 7.4), and 117 nM Fluorescein Was prepared. 75 mM potassium phosphate buffer (pH 7.4) was added to a standard substance (Trolox ™ : 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) to prepare 50 μM Trolox. A radical generator (AAPH: 2,2′-azo-bis (2-amidinopropane) dihydrochloride) was added with 75 mM potassium phosphate buffer (pH 7.4) to prepare 40 mM AAPH. To a 96-well fluorescent plate, 20 μl of sample (or 50 μM Trolox of standard substance) and 120 μl of 117 nM Fluorescein were added and pre-incubated at 37 ° C. for 15 minutes. Thereafter, 60 μl of 40 mM AAPH as a radical generator was added, and the fluorescence intensity was measured every minute at an excitation wavelength of 485 nm and a fluorescence wavelength of 520 nm.
一定の活性酸素種を発生させ、それによって分解される蛍光物質の蛍光強度を測定し、経時的に減少する蛍光強度の曲線を描いた場合、この反応系に抗酸化物質が共存すると蛍光物質の蛍光強度の減少速度が遅延する。この原理に基づき、検体(もしくは標準物質)存在下での蛍光強度の曲線下面積(AUC:Area Under the Curve)と、非存在下(ブランク)でのAUCとの差(net AUC)を算出する。検体のnet AUCについて、濃度既知の標準物質(Trolox)のnet AUCに対する相対値を求める。その相対値を基にTrolox濃度に換算して検体の抗酸化力とした。(単位μmol TE/g:マイクロモルTrolox当量/グラム) 〔analysis method〕
When the fluorescence intensity of a fluorescent substance that generates certain reactive oxygen species and is decomposed is measured and a fluorescence intensity curve that decreases over time is drawn, if an antioxidant is present in this reaction system, The rate of decrease in fluorescence intensity is delayed. Based on this principle, the difference (net AUC) between the area under the curve (AUC: Area Under the Curve) of the fluorescence intensity in the presence of the specimen (or standard substance) and the AUC in the absence (blank) is calculated. . For the net AUC of the specimen, the relative value of the standard substance (Trolox) having a known concentration with respect to the net AUC is obtained. Based on the relative value, it was converted into the Trolox concentration and used as the antioxidant power of the specimen. (Unit: μmol TE / g: micromol Trolox equivalent / gram)
チーズ酵素分解物の活性酸素吸収能力は、189.5±11.0μM Trolox当量/グラムであった。 〔result〕
The active oxygen absorption capacity of the cheese enzymatic degradation product was 189.5 ± 11.0 μM Trolox equivalent / gram.
(発酵乳の製造例1)
プレーンヨーグルトを、チーズ酵素分解物(明治チーズテイストコンセントレイトG(ゴーダタイプ))、L.bulgaricus JCM1002T、S.thermophilus ATCC19258により調製した。まず、脱脂粉乳10%培地を用いて、L.bulgaricus JCM1002T、S.thermophilus ATCC19258のバルクスターターを調製した。次に、ヨーグルトミックス(無脂乳固形分(SNF):9.5%、脂肪分(FAT):3.0%)にチーズ酵素分解物を混和し95℃で5分間加熱処理した。この加熱処理後のミックスに、L.bulgaricus JCM1002Tと、S.thermophilus ATCC19258のスターターを各1%で接種し、43℃、4時間で発酵して、プレーンヨーグルトを得た。このプレーンヨーグルトを冷蔵庫(5℃)で冷却してから、風味と物性を確認した。このとき、風味と物性はいずれも良好であった。 [Example 6] Production of fermented milk (Production Example 1 of fermented milk)
Plain yogurt was dissolved in a cheese enzyme digest (Meiji cheese taste concentrate G (Gouda type)), L. bulgaricus JCM1002 T , S. prepared by thermophilus ATCC 19258. First, using a skim milk powder 10% medium, L. bulgaricus JCM1002 T , S. A bulk starter of thermophilus ATCC 19258 was prepared. Next, the cheese enzyme degradation product was mixed with yoghurt mix (non-fat milk solids (SNF): 9.5%, fat (FAT): 3.0%), and heat-treated at 95 ° C for 5 minutes. In the mix after this heat treatment, L. bulgaricus JCM1002T; Thermophilus ATCC 19258 starter was inoculated at 1% each and fermented at 43 ° C. for 4 hours to obtain plain yogurt. The plain yogurt was cooled in a refrigerator (5 ° C.), and then the flavor and physical properties were confirmed. At this time, both flavor and physical properties were good.
(発酵乳の製造例2)
プレーンヨーグルトを、チーズ酵素分解物(明治チーズテイストコンセントレイトG(ゴーダタイプ))、血中尿酸値低減作用を有する乳酸菌であるL.gasseri OLL2959(NITE P-224)、L.bulgaricus JCM1002T、S.thermophilus ATCC19258により調製した。まず、脱脂粉乳10%培地を用いて、L.gasseri OLL2959、L.bulgaricus JCM1002T、S.thermophilus ATCC19258のバルクスターターを調製した。次に、ヨーグルトミックス(無脂乳固形分(SNF):9.5%、脂肪分(FAT):3.0%)にチーズ酵素分解物を混和し95℃で5分間加熱処理した。この加熱処理後のミックスに、L.bulgaricus JCM1002Tと、S.thermophilus ATCC19258のスターターを各1%、L.gasseri OLL2959のスターターを5%で接種し、43℃、4時間で発酵して、プレーンヨーグルトを得た。このプレーンヨーグルトを冷蔵庫(5℃)で冷却してから、風味と物性を確認した。このとき、風味と物性はいずれも良好であった。 [Example 7] Production of fermented milk (Production Example 2 of fermented milk)
Plain yogurt is a cheese enzyme degradation product (Meiji cheese taste concentrate G (Gouda type)), a lactic acid bacterium having a blood uric acid level reducing action. gasseri OLL2959 (NITE P-224), L.M. bulgaricus JCM1002 T , S. prepared by thermophilus ATCC 19258. First, using a skim milk powder 10% medium, L. gasseri OLL2959, L.M. bulgaricus JCM1002T, S. A bulk starter of thermophilus ATCC 19258 was prepared. Next, the cheese enzyme degradation product was mixed with yoghurt mix (non-fat milk solids (SNF): 9.5%, fat (FAT): 3.0%), and heat-treated at 95 ° C for 5 minutes. In the mix after this heat treatment, L. bulgaricus JCM1002 T ; thermophilus ATCC 19258
(プロセスチーズの製造例)
ニュージーランド産チェダーチーズを原料チーズとし、活性酸素吸収能力が約150μM Trolox当量/gのチーズ酵素分解物を添加して以下の製造方法によりプロセスチーズを調整した。配合割合は次の通りである。
〔配合表〕
ニュージーランド産チェダーチーズ 8.0kg
チーズ酵素分解物 0.05kg
溶融塩(トリポリリン酸ナトリウム) 0.2kg
水(加温のための蒸気を含む) 1.75kg
〔製造方法〕
チーズは予めミートチョッパで粗く粉砕しておく。20リットル容ケットル型ニーダーに原料を全て投入し(ただし、水の量は加温のための蒸気量を除いた量とする)、120rpmの回転数で撹拌しながら蒸気を吹き込み約10分間で85℃まで加温した。溶けて流動性を持つようになったチーズを200gずつ容器に採取し、密封して5℃の冷蔵庫で一晩冷却した。このものはチーズ酵素分解物の特異臭もなく、風味、組成は良好であった。 [Example 8] Production of process cheese (production example of process cheese)
New Zealand cheddar cheese was used as a raw material cheese, and an enzyme digestion product having an active oxygen absorption capacity of about 150 μM Trolox equivalent / g was added to prepare a processed cheese by the following production method. The blending ratio is as follows.
[Composition table]
New Zealand cheddar cheese 8.0kg
Cheese enzyme degradation product 0.05kg
Molten salt (sodium tripolyphosphate) 0.2kg
Water (including steam for heating) 1.75kg
〔Production method〕
The cheese is coarsely crushed beforehand with a meat chopper. All the raw materials are put into a 20-liter kettle type kneader (however, the amount of water is the amount excluding the amount of steam for heating), and steam is blown in at about 10 minutes while stirring at 120 rpm. Warmed to ° C. 200 g each of melted cheese having fluidity was collected in a container, sealed, and cooled overnight in a 5 ° C. refrigerator. This product did not have the specific odor of the enzyme degradation product of cheese and had good flavor and composition.
Claims (42)
- チーズ酵素分解物を有効成分とする血中尿酸値低下剤。 A blood uric acid level-lowering agent containing a cheese enzyme degradation product as an active ingredient.
- チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、請求項1記載の血中尿酸値低下剤。 The blood uric acid level lowering agent according to claim 1, wherein the cheese enzyme degradation product is a cheese enzyme degradation product having active oxygen absorption ability.
- チーズ酵素分解物の含有量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、請求項2記載の血中尿酸値低下剤。 3. The blood uric acid level-lowering agent according to claim 2, wherein the content of the cheese enzyme degradation product is an amount showing an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose.
- チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、請求項1~3のいずれか1項記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to any one of claims 1 to 3, wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase and protease.
- プロテアーゼが、エンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項4記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to claim 4, wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme.
- プロテアーゼが、エンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項4記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to claim 4, wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme.
- エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、請求項6記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to claim 6, wherein the endo-type protease is an acidic protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae.
- リパーゼが、ウシ由来のプレガストリックエステラーゼである、請求項4記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to claim 4, wherein the lipase is a bovine-derived pregastric esterase.
- チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、請求項1~3のいずれか1項記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to any one of claims 1 to 3, wherein the cheese enzyme degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria.
- 乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、請求項9記載の血中尿酸値低下剤。 Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. The blood uric acid level-lowering agent according to claim 9, which is lactis biovar diacylactis.
- 有効成分としてチーズ酵素分解物のみを含有する、請求項1~10のいずれか1項記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to any one of claims 1 to 10, comprising only a cheese enzyme degradation product as an active ingredient.
- チーズ酵素分解物および製剤補助剤のみを含有する、請求項1~11のいずれか1項記載の血中尿酸値低下剤。 The blood uric acid level-lowering agent according to any one of claims 1 to 11, comprising only a cheese enzyme degradation product and a formulation adjuvant.
- チーズ酵素分解物を有効成分とする活性酸素吸収剤。 An active oxygen absorbent containing a cheese enzyme degradation product as an active ingredient.
- 有効成分としてチーズ酵素分解物のみを含有する、請求項13記載の活性酸素吸収剤。 The active oxygen absorbent according to claim 13, comprising only a cheese enzyme degradation product as an active ingredient.
- 血中尿酸値を低下させるためのチーズ酵素分解物を含有する組成物。 A composition containing a cheese enzyme degradation product for lowering blood uric acid level.
- チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、請求項15記載の組成物。 The composition according to claim 15, wherein the cheese enzymatic degradation product is a cheese enzymatic degradation product having active oxygen absorption ability.
- チーズ酵素分解物の含有量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、請求項16記載の組成物。 The composition according to claim 16, wherein the content of the cheese enzyme degradation product is an amount exhibiting an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose.
- チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、請求項15~17のいずれか1項記載の組成物。 The composition according to any one of claims 15 to 17, wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with a lipase and a protease.
- プロテアーゼがエンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項18記載の組成物。 The composition according to claim 18, wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme.
- プロテアーゼがエンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項18記載の組成物。 The composition according to claim 18, wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme.
- エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、請求項20記載の組成物。 21. The composition according to claim 20, wherein the endo-type protease is an acidic protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae.
- リパーゼがウシ由来のプレガストリックエステラーゼである、請求項18記載の組成物。 The composition according to claim 18, wherein the lipase is a bovine-derived pregastric esterase.
- チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、請求項15~17のいずれか1項記載の組成物。 The composition according to any one of claims 15 to 17, wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria.
- 乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、請求項23記載の組成物。 Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. 24. The composition of claim 23, which is lactis biovar diacylactis.
- 有効成分としてチーズ酵素分解物のみを含有する、請求項15~24のいずれか1項記載の組成物。 The composition according to any one of claims 15 to 24, which contains only a cheese enzyme degradation product as an active ingredient.
- チーズ酵素分解物および製剤補助剤のみを含有する、請求項15~25のいずれか1項記載の組成物。 The composition according to any one of claims 15 to 25, which contains only a cheese enzyme degradation product and a formulation adjuvant.
- 活性酵素を吸収するためのチーズ酵素分解物を含有する組成物。 Composition containing cheese enzyme degradation product for absorbing active enzyme.
- 有効成分としてチーズ酵素分解物のみを含有する、請求項27記載の組成物。 28. The composition according to claim 27, which contains only cheese degradation products as an active ingredient.
- チーズ酵素分解物を投与することを特徴とする血中尿酸値の低下方法。 A method for lowering blood uric acid level, which comprises administering a cheese enzyme degradation product.
- チーズ酵素分解物が、活性酸素吸収能を有するチーズ酵素分解物である、請求項29記載の方法。 30. The method according to claim 29, wherein the cheese enzyme degradation product is a cheese enzyme degradation product having active oxygen absorption ability.
- チーズ酵素分解物の投与量が、1日投与量として50μM Trolox等量以上の活性酸素吸収能を示す量である、請求項30記載の方法。 The method according to claim 30, wherein the dose of the cheese enzyme degradation product is an amount showing an active oxygen absorption capacity of 50 μM Trolox equivalent or more as a daily dose.
- チーズ酵素分解物が、チーズをリパーゼ及びプロテアーゼで処理した分解物である、請求項29記載の方法。 30. The method according to claim 29, wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase and protease.
- プロテアーゼがエンド型プロテアーゼ、エキソ型プロテアーゼ、エキソ型ペプチダーゼ/エンド型プロテアーゼ複合酵素、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項32記載の方法。 The method according to claim 32, wherein the protease is at least two kinds selected from endo-type protease, exo-type protease, exo-type peptidase / endo-type protease complex enzyme, and protease / peptidase complex enzyme.
- プロテアーゼがエンド型プロテアーゼ、及びプロテアーゼ/ペプチダーゼ複合酵素から選ばれる2種以上である、請求項32記載の方法。 The method according to claim 32, wherein the protease is at least two selected from endo-type protease and protease / peptidase complex enzyme.
- エンド型プロテアーゼがAspergillus saitoi由来の酸性プロテアーゼであり、プロテアーゼ/ペプチダーゼ複合酵素がAspergillus oryzae由来のプロテアーゼ/ペプチダーゼ複合酵素である、請求項34記載の方法。 35. The method according to claim 34, wherein the endo-type protease is an acid protease derived from Aspergillus saitoi, and the protease / peptidase complex enzyme is a protease / peptidase complex enzyme derived from Aspergillus oryzae.
- リパーゼがウシ由来のプレガストリックエステラーゼである、請求項32記載の方法。 The method according to claim 32, wherein the lipase is a bovine-derived pregastric esterase.
- チーズ酵素分解物が、チーズをリパーゼ、プロテアーゼ及び乳酸菌で処理した分解物である、請求項29記載の方法。 30. The method according to claim 29, wherein the cheese enzymatic degradation product is a degradation product obtained by treating cheese with lipase, protease, and lactic acid bacteria.
- 乳酸菌がLactococcus lactis subsp.lactis、Lactococcus lactis subsp.cremoris、及びLactococcus lactis subsp.lactis biovar diacetylactisである、請求項37記載の方法。 Lactic acid bacteria are Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris and Lactococcus lactis subsp. 38. The method of claim 37, wherein Lactis biovar diacylactis.
- 有効成分として、実質的にチーズ酵素分解物のみを投与する、請求項29記載の方法。 30. The method according to claim 29, wherein substantially only a cheese enzyme degradation product is administered as an active ingredient.
- チーズ酵素分解物および製剤補助剤のみを投与する、請求項39の方法。 40. The method of claim 39, wherein only the cheese enzyme degradation product and the formulation adjuvant are administered.
- チーズ酵素分解物を投与することを特徴とする活性酸素吸収方法。 An active oxygen absorption method characterized by administering a cheese enzyme degradation product.
- 有効成分として、実質的にチーズ酵素分解物のみを投与する、請求項41記載の方法。 42. The method according to claim 41, wherein substantially only a cheese enzyme degradation product is administered as an active ingredient.
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CN201180009502.5A CN102753190B (en) | 2010-02-16 | 2011-02-14 | Uric acid level depressant in blood |
JP2012500581A JP5752108B2 (en) | 2010-02-16 | 2011-02-14 | Blood uric acid level lowering agent |
IN6608DEN2012 IN2012DN06608A (en) | 2010-02-16 | 2011-02-14 |
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Citations (4)
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JPH0641191A (en) * | 1992-03-04 | 1994-02-15 | Calpis Food Ind Co Ltd:The | Peptide and physiological activator containing the same |
WO2004047543A1 (en) * | 2002-11-28 | 2004-06-10 | Meiji Dairies Corporation | Functional foods and process for producing functional food |
WO2007080962A1 (en) * | 2006-01-13 | 2007-07-19 | Meiji Dairies Corporation | Anti-inflammatory composition and method for production of anti-inflammatory composition |
JP2009120519A (en) * | 2007-11-13 | 2009-06-04 | Meiji Milk Prod Co Ltd | Prophylactic or therapeutic agent for peptic ulcer |
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JPS59113869A (en) * | 1982-12-21 | 1984-06-30 | Snow Brand Milk Prod Co Ltd | Formation of cheese flavor having strong taste |
JPS61152235A (en) * | 1984-12-27 | 1986-07-10 | Meiji Milk Prod Co Ltd | Margarine having cheese taste and production thereof |
JPH0642814B2 (en) * | 1990-07-31 | 1994-06-08 | 不二製油株式会社 | Cheese manufacturing method |
US8263144B2 (en) * | 2005-11-17 | 2012-09-11 | Kraft Foods Global Brands Llc | Cheese flavor composition and process for making same |
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JPH0641191A (en) * | 1992-03-04 | 1994-02-15 | Calpis Food Ind Co Ltd:The | Peptide and physiological activator containing the same |
WO2004047543A1 (en) * | 2002-11-28 | 2004-06-10 | Meiji Dairies Corporation | Functional foods and process for producing functional food |
WO2007080962A1 (en) * | 2006-01-13 | 2007-07-19 | Meiji Dairies Corporation | Anti-inflammatory composition and method for production of anti-inflammatory composition |
JP2009120519A (en) * | 2007-11-13 | 2009-06-04 | Meiji Milk Prod Co Ltd | Prophylactic or therapeutic agent for peptic ulcer |
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CN102753190B (en) | 2016-11-23 |
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CN102753190A (en) | 2012-10-24 |
JPWO2011102310A1 (en) | 2013-06-17 |
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