EP0700433A1 - Procede d'hydrolyse de proteines - Google Patents

Procede d'hydrolyse de proteines

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Publication number
EP0700433A1
EP0700433A1 EP94913509A EP94913509A EP0700433A1 EP 0700433 A1 EP0700433 A1 EP 0700433A1 EP 94913509 A EP94913509 A EP 94913509A EP 94913509 A EP94913509 A EP 94913509A EP 0700433 A1 EP0700433 A1 EP 0700433A1
Authority
EP
European Patent Office
Prior art keywords
protein
proteolytic
preparation
hydrolysis
food product
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP94913509A
Other languages
German (de)
English (en)
Inventor
Per Munk Nielsen
Peter Hvass
Kim Hansen
Gitte Budolfsen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Novozymes AS
Original Assignee
Novo Nordisk AS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novo Nordisk AS filed Critical Novo Nordisk AS
Publication of EP0700433A1 publication Critical patent/EP0700433A1/fr
Withdrawn legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/58—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
    • C12N9/62—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi from Aspergillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01003—Triacylglycerol lipase (3.1.1.3)
    • 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
    • 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
    • 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
    • A23J3/342—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of collagen; of gelatin
    • 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
    • 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
    • A23J3/344—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins of casein
    • 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
    • A23J3/345—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of blood 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
    • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • 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
    • A23L15/00—Egg products; Preparation or treatment thereof
    • A23L15/25—Addition or treatment with microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20—Synthetic spices, flavouring agents or condiments
    • A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
    • 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
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/645—Proteins of vegetable origin; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/66—Enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21004—Trypsin (3.4.21.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21014—Microbial serine proteases (3.4.21.14)
    • 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
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/02—Preparations for cleaning the hair

Definitions

  • the present invention relates to a method of hydrolysing proteins, a protein hydrolysate obtained by the method, and food and non-food products containing the protein hydrolysate.
  • the present invention provides a method for hydrolysis of proteins by incubation with a proteolytic preparation derived from Aspergrillus oryzae and supplied by Novo Nordisk A/S, Denmark under the tradename FlavourzymeTM.
  • the invention provides protein hydrolysates obtained by the method of the invention.
  • the invention provides food and non-food products comprising a protein hydrolysate of the invention.
  • Fig. 1 shows the degree of hydrolysis (%DH) vs. time s of hydrolysis (hours) achieved by the method of the invention applied to sodium caseinate in the pH range of from pH 5 to pH 9 ( ⁇ : pH 5, A pH 6, O pH 7, D pH 8, • pH 9) ;
  • Fig. 2 shows the degree of hydrolysis (%DH) of soy protein isolate after 22 hours of hydrolysis according to the ⁇ o method of invention ( ⁇ : FlavourzymeTM, A : CorolaseTM 7092, O: CorolaseTM 7093, ⁇ : AlcalaseTM, D: NeutraseTM) ; and
  • Fig. 3 shows the degree of hydrolysis (%DH) of Na- caseinate after 22 hours of hydrolysis according to the method of invention ( ⁇ : FlavourzymeTM, A : CorolaseTM 7092, O: CorolaseTM is 7093, ⁇ : AlcalaseTM, D: NeutraseTM).
  • proteolytic enzyme components each of which may have any of the following approximate molecular weights: 23 kD, 27 kD, 31 kD, 32 kD, 35 kD, 38 kD, 42 kD, 47 kD, 53 kD, and 100 kD.
  • the present invention provides a method
  • proteolytic enzyme preparation which is derived from Aspergillus oryzae and comprises at least five proteolytic components having an approximate molecular weight, respectively, selected from 23 kD, 27 kD, 31 kD, 32 kD, 35 kD, 38 kD, 42 kD, 47 kD, 53 kD, and
  • the protein is incubated with a proteolytic preparation derived from Aspergillus oryzae and comprising at least five proteolytic components having the approximate molecular weights 23 kD, 31 kD, 35 kD, 38. kD, and
  • the molecular weight of the protease components in the proteolytic preparation was determined by using SDS polyacrylamide gel electrophoresis (SDS-PAGE) in a manner known to persons skilled in the art. In this way, the molecular weight of each protease component was determined.
  • the method of the invention is able to perform extensive protein hydrolysis of proteins, and the method leads to non-bitter hydrolysates and hydrolysates having pronounced soup-flavour/meat-flavour.
  • the extent of the protein hydrolysis may be determined by the degree of hydrolysis achieved.
  • the degree of hydrolysis (DH) is defined by the following formula:
  • the DH may be calculated according to Adler-Nissen ;
  • a DH which is higher than 35%, preferably higher than 60%, more preferably higher than 70%, most preferably higher than 80%.
  • the degree of protein solubility may be described by way of a
  • PSI Protein Solubility Index
  • the protein or proteinaceous material which may advantageously be hydrolysed by the present method may be any vegetable protein such as soy protein, grain proteins, e.g. wheat gluten or zein, rape seed protein, alfalfa protein, pea protein, fabaceous bean protein, cotton seed protein or sesame seed protein, or any animal protein or proteinaceous material such as milk protein, whey protein, casein, meat protein, fish protein, blood protein, egg white or gelatin.
  • vegetable protein such as soy protein, grain proteins, e.g. wheat gluten or zein, rape seed protein, alfalfa protein, pea protein, fabaceous bean protein, cotton seed protein or sesame seed protein
  • animal protein or proteinaceous material such as milk protein, whey protein, casein, meat protein, fish protein, blood protein, egg white or gelatin.
  • the proteolytic enzyme may suitably be added to the protein or proteinaceous material in an amount of 0.05-15 AU/100 g of protein, in particular 0.1-8 AU/100 g of protein.
  • the incubation may be performed at a pH from between about 4 and about 10, preferably between about 5 and about 9.
  • the method of the invention performs excellent even at extreme pH conditions, i.e. at pH values in all of the range of 5 to 9.
  • the incubation may be performed at any convenient temperature at which the enzyme preparation does not become inactivated, i.e. in the range of from about 20°C to about 70°C.
  • the proteo ⁇ lytic enzyme preparation may suitably be inactivated by increasing the temperature of the incubation mixture to above about 70°C, or by decreasing the pH of the incubation mixture to below about 4.0.
  • incubation of a protein or proteinaceous substrate may be performed with a combination of FlavourzymeTM and one or more other protease preparations.
  • Preferred protease preparations comprises neutral or alkaline proteases.
  • suitable neutral proteases are neutral proteases derived from Bacillus, preferably from Bacillus ⁇ ubtilis, such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename NeutraseTM.
  • alkaline proteases examples include alkaline proteases derived from Bacillus , preferably from Bacillus licheniformis, such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename AlcalaseTM and containing Subtilisin A (Subtilisin Carlsberg) as the active component.
  • AlcalaseTM alkaline proteases derived from Bacillus , preferably from Bacillus licheniformis, such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename AlcalaseTM and containing Subtilisin A (Subtilisin Carlsberg) as the active component.
  • the incubation carried out in the method of the invention may also be performed with a combination of FlavourzymeTM and one or more other lipase preparations.
  • Preferred lipase preparations comprises fungal lipases.
  • Suitable fungal lipases are lipases derived from Mucor, preferably from .Rhizojnucor miehei , such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename PalataseTMM; and lipases derived from Aspergillus , preferably from Aspergillus niger such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename PalataseTMA.
  • the present invention provides a protein hydrolysate obtained by the method of the invention.
  • the proteolytic activity may be determined with haemoglobin as substrate.
  • TCA trichloroacetic acid
  • a folder AF 4/5 describing the analytical method in more detail is available from Novo Nordisk A/S, DK-2880 Bagsvaerd, Denmark, upon request which folder is hereby included by reference.
  • a substrate for lipase was prepared by emulsifying glycerine tributyrat (MERCK) using gum-arabic as emulsifier. Lipase activity was assayed at pH 7 using pH stat method. One unit of lipase activity (LU/mg) is defined as the amount needed to liberate one micro ole fatty acid per minute.
  • Step 1 Centrifuge the fermentation supernatant, discard the precipitate. Adjust the pH of the supernatant to 7 and add gradually an equal volume of cold 96 % ethanol. Allow the mixture to stand for 30 minutes in an ice bath. Centrifuge and discard the precipitate.
  • Step 2 - Ion exchange chromatography. Filter the supernatant and apply on DEAE-fast flow (Pharmacia TM) column equilibrated with 50 mM tris-acetate buffer pH 7. Wash the column with the same buffer till absorption at 280 nm is lower than 0.05 OD. Elute the bound enzymatic activity with linear salt gradient in the same buffer (0 to 0.5 M NaCl ) using five column volumes, Pool the fractions containing enzymatic activity .
  • Step 3 Hydrophobic chromatography. Adjust the molarity of the pool containing enzymatic activity to 0.8 M by adding solid Ammonium acetate. Apply the enzyme on TSK gel Butyl- Toyopearl 650 C column (available from Tosoh Corporation Japan) which was pre-equilibrated with 0.8 M ammonium acetate. Wash the unbound material with 0.8 M ammonium acetate and elute the bound material with distilled water.
  • Step 4 Pool containing lipase activity is diluted with water to adjust conductance to 2 mS and pH to 7. Apply the pool on High performance Q Sepharose (Pharmacia) column pre- equilibrated with 50 mM tris -acetate buffer pH 7. Elute the bound enzyme with linear salt gradient.
  • the present invention relates to food products comprising a protein hydrolysate of the inven ⁇ tion.
  • the amount of protein hydrolysate incorporated in the food product will typically be in the range of 1-30% by weight.
  • An important food product of the present invention is an ingredient of a mother milk substitute for infants. Due to the high degree of hydrolysis obtained by the method of the invention, the protein hydrolysates of the invention may advan- tageously be incorporated in mother milk substitutes, the hydrolysate having a significantly lower allergenicity than unhydrolysed milk proteins.
  • the milk substitute may be for ⁇ mulated in substantially the same way as that indicated in the prior literature for products of this type (cf. for instance EP Patent Application 322,589) with the exception that the protein hydrolysate included in the known products is replaced by the protein hydrolysate of the present invention.
  • the food product of the invention may also include the protein hydrolysate of the invention as a protein supple- ment or to provide other properties of the food product.
  • the protein hydrolysate incorporated in the food product may for instance be based on meat or scrap meat (e.g. the so-called mechanically recovered meat, i.e. meat remaining on bones after the regular pieces of meat have been carved from animal carcases in the slaughterhouse.
  • meat or scrap meat e.g. the so-called mechanically recovered meat, i.e. meat remaining on bones after the regular pieces of meat have been carved from animal carcases in the slaughterhouse.
  • meat or scrap meat e.g. the so-called mechanically recovered meat, i.e. meat remaining on bones after the regular pieces of meat have been carved from animal carcases in the slaughterhouse.
  • mechanically recovered meat i.e. meat remaining on bones after the regular pieces of meat have been carved from animal carcases in the slaughterhouse.
  • Other proteinaceous by-products from the meat industry may
  • the resulting protein hydrolysate may then suitably be added to emulsified meat products, e.g. sausages or pates, or as flavour ingredients in soups or other food products.
  • a food product obtained from cow milk by the method of invention is preferably a cheese flavour product.
  • the extensive hydrolysis of the milk protein surprisingly leads to flavour compounds with very distinct cheese flavour.
  • Such cheese flavour products of the invention preferably find application in snack products, imitation cheese products, or as enhancers of cheese flavour in general.
  • the cheese flavour may be adjusted by use of lipases according to well established practice.
  • a traditional way of producing Hydrolysed Vegetable Protein (HVP) for use as flavour products is based on cooking in hydrochloric acid at a high temperature and for a long time. This is known to cause unwanted formation of chloric containing compounds. By the process of the invention it is now possible to obtain enzymatically produced HVP products, which are considered more healthy products.
  • the high DH obtainable by the process of the invention leads to desirable flavour characteristics and flavour enhancing properties.
  • the process of the invention may also be used for protein enrichment of dietetic products due to the non-bitter flavour obtained by the process.
  • Also advantageous is the very high protein solubility in a very broad pH range that is associated with the process of the invention. It has surprisingly been found that an excellent fermentation medium may be produced by subjecting milk to the method of the present invention. Such milk hydrolysates may be used for acceleration of the fermentation in yoghurt production or in the production of lactic acid starter bacteria cultures, by addition in relatively small amounts. The shortening of process time increases production capacity and reduces the risk of infection, e.g. bacteriophage infection.
  • the method of the invention may be used in combination with a fermentation process, preferably a fermentation process for the production of a food product.
  • the proteolytic enzyme preparation e.g. FlavourzymeTM
  • the protein hydrolysate of the invention or both may be added in a fermentation process, preferably a food fermentation process, for enhancing the productivity of the fermentation process.
  • fermentation processes are processes involving or fermenting fish (fish sauce) , cocoa beans or soy (such as soy sauce, tempeh, iso) .
  • the addition of the proteolytic enzyme preparation or the addition of protein hydrolysate may be useful for decreasing the total process time, i.e. increasing the rate of fermentation.
  • Non-food products e.g. FlavourzymeTM
  • the process of the invention may also find application in the non-food area.
  • the flavour characteristics obtained by the method of invention may advantageously be used for production 5 of pet food.
  • Preparation B 1% FlavourzymeTM, 2% NeutraseTM 0.5L, and 0.15% AlcalaseTM 2,4L.
  • FlavourzymeTM is a proteolytic preparation derived from
  • NeutraseTM 0.5L is a proteolytic preparation derived from Bacillus subtilis
  • AlcalaseTM 2.4L is a proteolytic preparation derived from Bacillus licheniformis .
  • hydrolysates were served before a tasting panel in a 4% solution.
  • the meat flavour was most pronounced in hydrolysates obtained with Preparation A.
  • flavour-samples 1, 2, and 3 were now diluted in cream cheese, and a fourth sample was prepared as a control.
  • sample No. 4.1 6.0 g of fresh milk + 20 g of cheese.
  • Sample No. 1.1 (FlavourzymeTM) appeared to have a very strong and intense flavour and a taste of well ripened cheese.
  • Sample No. 2.1 (FlavourzymeTM + PalataseTM) appeared to have a 0 well balanced cheese flavour.
  • sample No. 3.1 turned out to give the weakest taste impression.
  • sample No. 4.1 was evaluated being sour/sourish.
  • PalataseTM creates a new and much more simple way of producing cheese flavour.
  • the hydrolysis was carried out for 20 hours at 50°C. Then the enzymes were denatured by heat treatment at 80°C for 5 min.
  • Skim milk was heat treated at 90°C for 2 min and three samples of 200 ml were prepared.
  • the temperature was adjusted to 41°C, and yoghurt culture YC DVS from Chr. Hansens Lab. was added to all three samples in a level of 10 6 bacteria per g.

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Abstract

L'invention concerne un procédé d'hydrolyse d'une protéine végétale ou animale par incubation avec une préparation à base d'enzymes protéolytiques dérivée de l'Aspergillus oryzae telle que la FlavourzymeTM comprenant au moins cinq composés protéolytiques présentant une masse molaire approximative de 23 kD, 27 kD, 31 kD, 32 kD, 38 kD, 42 kD, 47 kD, 53 kD ou de 100 kD, qui permet de produire un hydrolysat protéique utile dans ou comme un produit alimentaire tel que le lait maternel de substitution, le fromage, la protéine végétale hydrolysée, un aromatisant et un adjuvant de procédé pour la fermentation de produits alimentaires ou un produit non alimentaire tel qu'un aliment pour animaux familiers, des cosmétiques. Le procédé selon l'invention permet d'obtenir un degré d'hydrolyse et de goût élevés et solubilité protéique élevée.
EP94913509A 1993-04-26 1994-04-25 Procede d'hydrolyse de proteines Withdrawn EP0700433A1 (fr)

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DK93467A DK46793D0 (da) 1993-04-26 1993-04-26 Enzym
DK467/93 1993-04-26
PCT/DK1994/000165 WO1994025580A1 (fr) 1993-04-26 1994-04-25 Procede d'hydrolyse de proteines

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DK (1) DK46793D0 (fr)
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WO (1) WO1994025580A1 (fr)

Families Citing this family (69)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE257329T1 (de) * 1994-10-26 2004-01-15 Novozymes As Verfahren für die herstellung eines milch-protein-hydrolysates
DE19632455C1 (de) * 1996-08-12 1997-08-21 Cpc Maizena Gmbh Verfahren zur Herstellung eines Proteinhydrolysats aus proteinhaltigen tierischen Produkten
EP0936875B1 (fr) * 1996-10-30 2003-06-25 Novozymes A/S Procede de preparation d'un agent aromatisant alimentaire
AU732747B2 (en) * 1996-10-30 2001-04-26 Novozymes A/S Method of producing food flavoring agent
ES2116937B1 (es) * 1996-11-29 1999-03-01 Consejo Superior Investigacion Procedimiento de obtencion de peptonas vegetales de alto grado de hidrolisis y sus aplicaciones.
DK0946106T3 (da) * 1996-12-23 2002-09-16 Dsm Nv Fremgangsmåde til fremstilling af et proteinhydrolysat
CA2275423A1 (fr) * 1996-12-23 1998-07-02 Dsm N.V. Renforcateur de gout
US6007851A (en) * 1996-12-23 1999-12-28 Gist-Brocades, B.V. Process for producing a flavor enhancer
WO1998051163A2 (fr) * 1997-05-16 1998-11-19 Novo Nordisk Biotech, Inc. Procedes de production d'hydrolysats proteiques
AU7381098A (en) 1997-05-16 1998-12-08 Novo Nordisk Biotech, Inc. Polypeptides having prolyl pipeptidyl aminopeptidase activity and nucleic acids encoding same
CA2421058C (fr) 1999-10-20 2010-04-20 Jon Bragi Bjarnason Hydrolysats proteiques obtenus a partir de proteases marines
FR2804691B1 (fr) 2000-02-04 2003-11-07 Roquette Freres Composition azotee resultant de l'hydrolyse du gluten de mais et son procede de fabrication
MY127310A (en) * 2000-04-28 2006-11-30 Sumitomo Rubber Ind Deproteinizing agent, deproteinized natural rubber latex using the same, and method of producing rubber product
AU2001265935A1 (en) * 2000-06-23 2002-01-14 Societe Des Produits Nestle S.A. Production of biohydrolysates
KR20020026640A (ko) * 2000-10-02 2002-04-12 김강권 항원성이 저감된 카제인 가수분해물의 제조방법 및 이가수분해물을 함유하는 유제품
AU1142001A (en) 2000-10-19 2002-04-29 Dsm N.V. Protein hydrolysates
CN1234288C (zh) * 2001-03-05 2006-01-04 科学与工业研究委员会 从大豆粉中制备蛋白质水解产物的方法
WO2002069734A1 (fr) * 2001-03-05 2002-09-12 Council Of Scientific And Industrial Research Procede de preparation d'hydrolysat proteique a partir des proteines du lait
KR20030028089A (ko) * 2001-09-27 2003-04-08 주식회사 태훈바이오 홍게 가공부산물을 이용한 반응향미제 및 이의 제조방법
WO2003054186A1 (fr) * 2001-12-21 2003-07-03 Dsm Ip Assets B.V. Nouvelles presures
FR2836014B1 (fr) * 2002-02-15 2004-07-23 Gervais Danone Sa Nouveau procede de fabrication de produits laitiers fermentes mettant en oeuvre des enzymes d'origine bacterienne
US6835385B2 (en) * 2002-06-14 2004-12-28 Carol J. Buck Compositions and methods for softening, thinning and removing hyperkeratotic tissue
US6787168B1 (en) * 2003-03-18 2004-09-07 James W. Sawhill Peptide product
US7785824B2 (en) 2003-05-05 2010-08-31 Conopco Inc. Hydrolysed casein product comprising tripeptides IPP and/or VPP
ITMI20031528A1 (it) * 2003-07-25 2005-01-26 Ct Sperimentale Del Latte S P A Terreno di fermentazione a base di un substrato proteico altamente idrolizzato
KR101174299B1 (ko) * 2003-08-01 2012-08-16 칼피스가부시키가이샤 카세인 가수분해물, 그 제조법 및 그 용도
US7485323B2 (en) 2005-05-31 2009-02-03 Gelita Ag Process for making a low molecular weight gelatine hydrolysate and gelatine hydrolysate compositions
WO2007066841A1 (fr) * 2005-12-09 2007-06-14 Serombio Co., Ltd. Hydrolisat de levure renfermant du cyclo-his-pro et procede de production afferent
JP4726129B2 (ja) * 2006-01-17 2011-07-20 学校法人北里研究所 ビフィズス菌増殖促進ペプチド
NZ544925A (en) * 2006-01-24 2009-02-28 Vital Food Processors Ltd Method of preparing a protein rich liquid extract from a protein source using a thiol cysteine protease aka actinidin from kiwifruit
WO2008066715A2 (fr) 2006-11-22 2008-06-05 Standard Biologics, Inc. Procédé de traitement utilisant une protéase d'aspergillus oryzae
EP1969950A1 (fr) * 2007-03-12 2008-09-17 Cargill, Incorporated Protéine de céréales partiellement hydrolysées
FR2925325B1 (fr) * 2007-12-21 2010-02-26 Vincience Utilisation d'un hydrolysat de colza en tant que principe actif activateur de la synthese des aquaporines
DK2300606T3 (da) * 2008-06-03 2012-04-16 Novozymes As Fremgangsmåde til fremstilling af et kaseinhydrolysat
CN102046022A (zh) * 2008-06-03 2011-05-04 诺维信公司 产生小麦蛋白水解物的方法
US20110165305A1 (en) * 2008-07-18 2011-07-07 Novozymes A/S Method for preparing a protein hydrolysate
WO2010074955A1 (fr) 2008-12-16 2010-07-01 Novozymes, Inc. Polypeptides ayant une activité carboxypeptidase et polynucléotides les codant
CN101518285B (zh) * 2009-03-30 2011-07-20 山东轻工业学院 一种干酪的生产方法
EP2258208A1 (fr) * 2009-06-02 2010-12-08 University of Limerick Produit de protéine avec une immunogénicité modifiée
BR112012016232A2 (pt) * 2009-12-30 2017-03-07 Solae Llc processo para separação e purificação de um produto de kti
NO20100370A1 (no) 2010-03-08 2011-09-09 Marine Bioproducts As Peptidmateriale, fôrsammensetninger og preparater, og anvendelser derav.
AU2011341063B2 (en) * 2010-12-06 2016-03-03 Cargill, Incorporated Process for liquefying cereal proteins
CN102550895A (zh) * 2010-12-28 2012-07-11 雀巢公司 来自于食物固态发酵的酶制剂
EP2489281A1 (fr) * 2011-02-17 2012-08-22 University of Limerick Hydrolysat de caséine
WO2012121612A1 (fr) * 2011-03-04 2012-09-13 Meat Biologics Research Limited Procédé de fabrication d'une composition comestible
WO2012121611A2 (fr) * 2011-03-04 2012-09-13 Meat Biologics Research Limited Composition comestible et son procédé de fabrication
UA111078C2 (uk) * 2011-05-03 2016-03-25 Нестек С.А. Гідролізат білкового субстрату і спосіб його виготовлення
ES2395982B1 (es) * 2011-07-08 2014-01-22 Apc Europe S.A. Suplemento alimenticio, método para incrementar la masa bacteriana en el rumen de un rumiante y preparado alimenticio y usos correspondientes.
US20140242215A1 (en) * 2011-10-03 2014-08-28 Specialites Pet Food Animal digests having enhanced palatability for use in pet food
PL2948003T3 (pl) 2013-01-23 2021-01-25 Bottled Science Limited Kompozycja napoju poprawiająca skórę
US9139458B2 (en) 2013-03-15 2015-09-22 Janet Angel Compositions and methods of use
JP6207922B2 (ja) * 2013-08-14 2017-10-04 マルハニチロ株式会社 長期常温保存可能なレバ刺し状食品
CN103549435A (zh) * 2013-11-19 2014-02-05 哈尔滨艾克尔食品科技有限公司 乳清蛋白肽口服液的制作方法
JP6481923B2 (ja) * 2014-05-02 2019-03-13 奥野製薬工業株式会社 畜肉改良剤
WO2015181917A1 (fr) * 2014-05-28 2015-12-03 天野エンザイム株式会社 Hydrolysat d'albumine fortement émulsifiable
EP3148352A1 (fr) * 2014-05-29 2017-04-05 Lb Lyopharm S.R.L. Procédé de préparation de protéines végétales fonctionnelles
AU2015357195B2 (en) * 2014-12-01 2019-06-13 Novozymes A/S Method for producing a protein hydrolysate
CN107404922B (zh) 2015-03-30 2021-09-14 雀巢产品有限公司 乳基蛋白质水解物及由其制备的组合物
CN104719920B (zh) * 2015-04-09 2017-03-08 广州超亚化妆品科技有限公司 多肽氨基酸混合物及其制备方法与应用
CN105077158B (zh) * 2015-08-24 2018-05-01 王婧婧 一种芝士风味乳香基的制备方法
DE102017200434A1 (de) 2016-01-13 2017-07-13 Robert Finke Molke-haltiges Körperpflegeprodukt
CN107772077A (zh) * 2016-08-30 2018-03-09 中粮营养健康研究院有限公司 一种菜籽饼和提高菜籽饼的蛋白质溶解度的方法
CN107788223A (zh) * 2016-08-30 2018-03-13 中粮营养健康研究院有限公司 一种提高菜籽饼的蛋白质溶解度的方法
WO2018053197A2 (fr) * 2016-09-14 2018-03-22 Glanbia Nutritionals (Ireland) Ltd. Produits protéiques agglomérés et leur procédé de production
CN106616995A (zh) * 2016-12-27 2017-05-10 济南邦地生物工程有限公司 一种健脑口服液及其生产方法
US20220061353A1 (en) * 2017-11-03 2022-03-03 Cargill, Incorporated Pea protein hydrolysate
CN108771194A (zh) * 2018-07-02 2018-11-09 广东海洋大学 一种利用耐高温地衣芽孢杆菌协助酶解水产蛋白制备海鲜调味包的方法
CN110810687B (zh) * 2019-11-21 2022-08-23 江南大学 一种利用内源性内肽酶和外肽酶水解芝麻蛋白制备低聚肽清凉饮料的方法
CN116828989A (zh) * 2021-02-04 2023-09-29 天野酶制剂(美国)有限公司 天然干酪的制造方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3306009C2 (de) * 1983-02-22 1994-02-17 Roehm Gmbh Verfahren zur Herstellung von Proteinhydrolysaten
CN86108557A (zh) * 1986-12-20 1988-07-13 同济大学 以含蛋白质的废料为原料酶法提取蛋白质水解物的方法及其产品
CH679542A5 (fr) * 1989-11-27 1992-03-13 Nestle Sa

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9425580A1 *

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CN1090675C (zh) 2002-09-11
NZ265247A (en) 1996-07-26
JPH08509366A (ja) 1996-10-08
AU6564094A (en) 1994-11-21
KR960701993A (ko) 1996-03-28
AU681653B2 (en) 1997-09-04
CN1121732A (zh) 1996-05-01
WO1994025580A1 (fr) 1994-11-10

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