JP7339261B2 - 低アレルゲン性の乳児用調製乳及びそれを調製する方法 - Google Patents
低アレルゲン性の乳児用調製乳及びそれを調製する方法 Download PDFInfo
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- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
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- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 235000020978 long-chain polyunsaturated fatty acids Nutrition 0.000 description 1
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- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
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- 239000011733 molybdenum Substances 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229940054441 o-phthalaldehyde Drugs 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 229960002446 octanoic acid Drugs 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 1
- 235000020665 omega-6 fatty acid Nutrition 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000003346 palm kernel oil Substances 0.000 description 1
- 235000019865 palm kernel oil Nutrition 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 235000020615 partially hydrolysed formula Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- ZWLUXSQADUDCSB-UHFFFAOYSA-N phthalaldehyde Chemical compound O=CC1=CC=CC=C1C=O ZWLUXSQADUDCSB-UHFFFAOYSA-N 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 235000013406 prebiotics Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- RADKZDMFGJYCBB-UHFFFAOYSA-N pyridoxal hydrochloride Natural products CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 150000003839 salts Chemical group 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000008347 soybean phospholipid Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000002600 sunflower oil Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 229960003080 taurine Drugs 0.000 description 1
- 238000007669 thermal treatment Methods 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 125000003203 triacylglycerol group Chemical group 0.000 description 1
- 150000003626 triacylglycerols Chemical group 0.000 description 1
- PHYFQTYBJUILEZ-IUPFWZBJSA-N triolein Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(OC(=O)CCCCCCC\C=C/CCCCCCCC)COC(=O)CCCCCCC\C=C/CCCCCCCC PHYFQTYBJUILEZ-IUPFWZBJSA-N 0.000 description 1
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/14—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment
- A23C9/142—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration
- A23C9/1422—Milk preparations; Milk powder or milk powder preparations in which the chemical composition of the milk is modified by non-chemical treatment by dialysis, reverse osmosis or ultrafiltration by ultrafiltration, microfiltration or diafiltration of milk, e.g. for separating protein and lactose; Treatment of the UF permeate
-
- 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
- A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/015—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1322—Inorganic compounds; Minerals, including organic salts thereof, oligo-elements; Amino-acids, peptides, protein-hydrolysates or derivatives; Nucleic acids or derivatives; Yeast extract or autolysate; Vitamins; Antibiotics; Bacteriocins
-
- 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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- 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
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
- A23L33/19—Dairy proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
- A23J3/343—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2300/00—Processes
- A23V2300/28—Hydrolysis, degree of hydrolysis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Pediatric Medicine (AREA)
- Microbiology (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- Water Supply & Treatment (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Dairy Products (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
Description
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、乳タンパク質の部分的加水分解物を準備する工程、
(ii)この部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
(iii)この清浄化された部分的加水分解物を任意選択的に濃縮する工程、
並びに
(iv)この(濃縮された)清浄化された部分的加水分解物を、CMAを発症するリスクがある乳児用の栄養組成物へと製剤化する工程
を含む方法を提供する。
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、乳タンパク質の部分的加水分解物を準備する工程、
(ii)この部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
(iii)この清浄化された部分的加水分解物を任意選択的に濃縮する工程、
並びに
(iv)この(濃縮された)清浄化された部分的加水分解物を、CMAを発症するリスクがある乳児用の栄養組成物へと製剤化する工程
を含み、
清浄化する工程(ii)は、10~100kDaの範囲の分子カットオフを有する膜を使用する、この部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含むろ液を回収することを含む、
方法を提供する。
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、乳タンパク質の部分的加水分解物を準備する工程、
(ii)この部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
(iii)この清浄化された部分的加水分解物を任意選択的に濃縮する工程、
並びに
(iv)この(濃縮された)清浄化された部分的加水分解物を、CMAを発症するリスクがある乳児用の栄養組成物へと製剤化する工程
を含み、
清浄化する工程(ii)は、この部分的加水分解物のサイズ排除クロマトグラフィー、及び清浄化された部分的加水分解物を含むろ液を回収することを含む、
方法を提供する。
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、この乳タンパク質の部分的加水分解物を準備する工程、
並びに
(ii)この部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
並びに
(iii)この清浄化された部分的加水分解物を任意選択的に濃縮する工程
を含み、
清浄化する工程(ii)は、10~100kDaの範囲の分子カットオフを有する膜を使用する、この部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含むろ液を回収することを含むか、又は
清浄化する工程(ii)は、この部分的加水分解物のサイズ排除クロマトグラフィー、及び清浄化された部分的加水分解物を含むろ液を回収することを含む
方法に関する。
この実施例は、部分的乳タンパク質加水分解物の様々なサイズの画分への分画を示す。
S8x、S6-50、及びS375は、ホエイタンパク質濃縮物(WPC)から得られ且つ加水分解度が約9.2%である、噴霧乾燥された部分的加水分解物である。
S100は、WPCから得られ且つ加水分解度が約15%である、噴霧乾燥された広範囲加水分解物である。
4種の異なる部分的乳タンパク質加水分解物を、50mg/mLのタンパク質濃度にてMilliQに溶解させた。振盪下での15分にわたる溶解後、これらの試料を遠心分離した(2000g、10分、20℃)。その後、試料を15分にわたり4℃でインキュベートし、分離した水層及び脂肪層の形成を可能にした。水層を注意深く取り出し、脂肪層を廃棄した。残余のペレットを、PBS、Tris/尿素(0.05M/6M)、又は追加のジチオエリトリトール(DTE、5mg/mL)を含むTris/尿素(0.05M/6M)のいずれかの固定量に溶解させた。得られた画分のタンパク質濃度を、Nanodropシステム(ThermoScientific)を使用して、280nmでの吸収測定により決定した。
分画した試料の様々な画分を、サイズ排除クロマトグラフィー(SEC)分析にかけた。粒径が1.7μmであり孔径が200ÅであるWaters Acquity UPLCタンパク質カラムを使用した。PBS(100mM、pH6.8+150mMのNaCl)による均一濃度流(isocratic flow)(0.3mL/分)を適用した。試料10μLを注入し、214nmでのUV吸光度によりタンパク質を検出した。
実験用の乳加水分解物を、吸光度及び蛍光の測定にかけた。試料を、MilliQでタンパク質5.0又は2.5mg/mLまで希釈した。試料を、96ウェルプレートのウェルにピペットで入れた(低結合、100μL/ウェル)。294nm及び490nmで吸光度を測定した。
sRAGE阻害アッセイ
Liu他(Liu F,et al.Food Funct.2016 Jan;7(1):239-49;Liu F,et al.,J Agric Food Chem.2016 Aug 24;64(33):6477-86)により以前に説明されているように、様々な乳タンパク質加水分解物のsRAGE結合能力を、競合アッセイにより評価した。
乳アレルギー患者からのヒト血清
乳アレルギー患者からのヒト血清を、Rijnstate Hospital(Arnhem,The Netherlands)から得た。3例の異なる患者の等量をプールした。それぞれの患者に関して、特異的IgEレベルを決定した。決定した値を表1に示す。
スピン遠心分離により、ヒト血清からIgGを除去した。プロトコルに従って、アガロースに結合したプロテインGを平衡化して、15mL採取管に入れた。ヒト血清を結合緩衝液で2倍に希釈し、カラム中のアガロース樹脂上にロードし、10分にわたりインキュベートした。その後、このカラムを遠心分離し(1000g、1分)、フロースルーを保管した。このカラムに追加の結合緩衝液を添加し、続いて遠心分離することにより、このカラムを3回洗浄した。タンパク質濃度に基づいて、関連する洗浄画分をプールした。溶出緩衝液を添加することにより、このカラムから、結合したIgGを溶出させ、その後に廃棄した。
ヒトIgEレセプター(FcERI)のアルファサブユニットがトランスフェクトされたラット好塩基球白血病(Rat Basophil Leukemia)(RBL)細胞を用いて、様々な乳加水分解物画分の脱顆粒誘発能力を評価した。細胞を、37℃、5%CO2にて、5%の熱不活性化ウシ胎児血清(FBS)、1%のペニシリン、1%のストレプトマイシン、及び1%のグルタミンを含むMEM中で増殖させた。細胞をMEMで洗浄し、フラスコの底から掻き取った。細胞を、遠心分離(400g、5分、RT)及びMEMへの再懸濁により、MEMで2回洗浄した。その後、細胞を、MEM+1%のグルタミン中に7.5×104/ウェルの密度で、96ウェルプレートに播種した。
結果を図4に示す。例示的な加水分解物S8xの画分のそれぞれは、分子量が>100kDaである画分を除いて、WPCと比べて有意に少ないRBL放出を誘発した。この画分は最大のRBL放出を示しており、WPCにより誘発された放出に対する54%減少に等しい。対照的に、分子量がより低い画分は、より低いβ-ヘキソサミニダーゼ放出を引き起こし、分子量が<3kDaであるタンパク質を含む画分の場合のWPCの14%まで低下した。ペレット由来の不溶性タンパク質は、RBL放出にほとんど寄与しなかった。
部分的加水分解物の調製
ホエイタンパク質濃縮物(WPC)又はカゼインを含む乳タンパク質を、50~60℃で、10~15%(重量/重量)の最終タンパク質濃度まで水に溶解させる。このタンパク質溶液のpHを、45%の水酸化カリウム(KOH)又は水酸化ナトリウム(NaOH)でpH6~8に調整する。このタンパク質溶液に、1:50~1:500の酵素/基質(E/S)比でエンドペプチダーゼ(アルカラーゼ(Alcalase))及びエキソペプチダーゼ(フレーバーザイム(Flavourzyme))を添加することにより、このタンパク質の加水分解を開始させる。この反応混合物を、4~24時間にわたり50~60℃で保持する。この酵素反応を、この反応混合物を90℃まで加熱することにより停止させ、その後、この混合物を少なくとも10分にわたり90℃で保持する。加熱後、この混合物を10℃まで冷却する。
部分的加水分解物を、100kDaの分子量カットオフを有するAmicon Centrifugal Filter Unit(Millipore)を使用して、10℃にて膜でろ過する(10分、3363g、RT)。保持液を水に再懸濁させ、透過液中において280nmで検出されるタンパク質がなくなるまで遠心分離工程を繰り返す。清浄化された加水分解物を含む透過液画分をプールし、更なる下流の処理にかける。
清浄化された加水分解物を、60mbarの真空圧を適用して30℃にて回転膜蒸発を使用して、40~60%の乾物含量まで濃縮する。濃縮された加水分解物を、200℃の入口温度及び80℃の出口温度を適用して、Buechiベンチトップ噴霧ドライヤ(Buechi bench-top spray drier)を使用して噴霧乾燥させる。
清浄化されて乾燥された部分的加水分解物を、表2に示す最終濃度まで、生後0~6ヶ月用の乳児用調製乳へと製剤化する。味が良好な乳児用調製乳は、清浄化されたホエイ又はカゼインタンパク質加水分解物6~16重量%、脂肪成分18~29重量%、炭水化物含有量<60重量%、プレバイオティクス成分3~8重量%を(製品100g当たり)含む。ミネラル、微量元素、及びビタミンのような更なる成分を、法律で推奨される量で組み入れる。
Claims (18)
- 牛乳アレルギー(CMA)を発症するリスクがある乳児用の低アレルゲン性栄養組成物を生成する方法であって、
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、乳タンパク質の部分的加水分解物を準備する工程、ここで、該部分的加水分解物は、加水分解度(DH)が5~20%の範囲である、
(ii)前記部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
(iii)前記清浄化された部分的加水分解物を任意選択的に濃縮する工程、
並びに
(iv)前記(濃縮された)清浄化された部分的加水分解物を、CMAを発症するリスクがある乳児用の栄養組成物へと製剤化する工程
を含み、
前記清浄化する工程(ii)は、10~100kDaの範囲の分子カットオフを有する膜を使用する、前記部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含むろ液を回収することを含むか、又は
前記清浄化する工程(ii)は、前記部分的加水分解物のサイズ排除クロマトグラフィー、及び清浄化された部分的加水分解物を含む前記ろ液を回収することを含む、
方法。 - 前記清浄化する工程(ii)は、10~100kDの範囲の分子カットオフを有する膜を使用する、前記部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含む前記ろ液を回収することを含む、請求項1に記載の方法。
- 前記部分的加水分解物は、ホエイタンパク質、酸ホエイタンパク質、スイートホエイタンパク質、ホエイタンパク質濃縮物、ホエイタンパク質単離物、脱塩ホエイ粉末、及びカゼイン塩からなる群から選択される1種又は複数種の牛乳タンパク質から得られる、請求項1又は2に記載の方法。
- 前記部分的加水分解物は、部分的ホエイタンパク質加水分解物を含むか、又は部分的ホエイタンパク質加水分解物であり、前記部分的加水分解物は、部分的ベータ-ラクトグロブリン加水分解物及び/若しくは部分的アルファ-ラクトアルブミン加水分解物を含むか、又は部分的ベータ-ラクトグロブリン加水分解物及び/若しくは部分的アルファ-ラクトアルブミン加水分解物であり、前記部分的加水分解物は、部分的カゼイン加水分解物を含むか、又は部分的カゼイン加水分解物である、請求項3に記載の方法。
- 前記部分的加水分解物は、加水分解度(DH)が6~18%の範囲である、請求項1~4のいずれか一項に記載の方法。
- 前記部分的加水分解物は、加水分解度(DH)が7~11%の範囲である、請求項5に記載の方法。
- 前記部分的加水分解物は、少なくとも10重量%の、サイズが5kDa以上であるペプチドと、少なくとも15重量%の、サイズが1~5kDaの範囲のペプチドとを含む、請求項1~6のいずれか一項に記載の方法。
- 前記部分的加水分解物中の前記ペプチドの前記サイズ分布は、部分的加水分解物中に存在するペプチドの乾燥重量を基準として、40~60% <1kDa、10~14% 1~<2kDa、8~16% 2~<5kDa、3~7% 5~<10kDa、及び8~12% >10kDaである、請求項1~7のいずれか一項に記載の方法。
- 10~50kDaの範囲の分子カットオフを有する膜の使用を含む請求項1~8のいずれか一項に記載の方法。
- 10~20kDaの範囲の分子カットオフを有する膜の使用を含む請求項9に記載の方法。
- ろ過は、精密ろ過(MF)、限外ろ過(UF)、炭素ろ過、又はポリッシュろ過を含む、請求項1~10のいずれか一項に記載の方法。
- 前記清浄化された部分的加水分解物を濃縮する工程(iii)を含む、請求項1~11のいずれか一項に記載の方法。
- 工程(iv)は、前記濃縮された、清浄化された部分的加水分解物と、炭水化物の供給源、脂質及び通常のビタミンの供給源、微量元素、並びにミネラルとを組み合わせることを含む、請求項1~12のいずれか一項に記載の方法。
- 工程(iv)は、ガラクトオリゴ糖(GOS)、ヒト乳オリゴ糖(HMO)、及びTGFβからなる群から選択される少なくとも1種の成分を含めることを含む、請求項1~13のいずれか一項に記載の方法。
- 乳タンパク質の部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する方法であって、
(i)水性媒体中に1種又は複数種の牛乳タンパク質を含む出発組成物を酵素処理にかけることにより得られる、前記乳タンパク質の部分的加水分解物を準備する工程、ここで、該部分的加水分解物は、加水分解度(DH)が5~20%の範囲であり、少なくとも10重量%の、サイズが5kDa以上であるペプチドと、少なくとも15重量%の、サイズが1~5kDaの範囲のペプチドとを含む
並びに
(ii)前記部分的加水分解物を、RAGE結合し得る及び/又は好塩基球脱顆粒誘発能力を有する1種又は複数種の成分から清浄化する工程、
並びに
(iii)前記清浄化された部分的加水分解物を任意選択的に濃縮する工程
を含み、
前記清浄化する工程(ii)は、10~100kDaの範囲の分子カットオフを有する膜を使用する、前記部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含むろ液を回収することを含むか、又は
前記清浄化する工程(ii)は、前記部分的加水分解物のサイズ排除クロマトグラフィー、及び清浄化された部分的加水分解物を含む前記ろ液を回収することを含む
方法。 - 前記清浄化する工程(ii)は、10~100kDの範囲の分子カットオフを有する膜を使用する、前記部分的加水分解物のろ過、及び清浄化された部分的加水分解物を含むろ液を回収することを含む、請求項15に記載の方法。
- 前記膜が10~50kDaの範囲の分子カットオフを有する、請求項16に記載の方法。
- 前記膜が10~20kDaの範囲の分子カットオフを有する、請求項17に記載の方法。
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DK3740087T3 (da) | 2022-08-08 |
ES2923576T3 (es) | 2022-09-28 |
BR112020012263A2 (pt) | 2020-11-24 |
KR20200108829A (ko) | 2020-09-21 |
EP3740087A1 (en) | 2020-11-25 |
JP2021510502A (ja) | 2021-04-30 |
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US11903392B2 (en) | 2024-02-20 |
RU2020124227A3 (ja) | 2022-04-11 |
AU2019209384A1 (en) | 2020-05-21 |
CN111726994A (zh) | 2020-09-29 |
US20200345024A1 (en) | 2020-11-05 |
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