AU6564094A - A method for hydrolysing proteins - Google Patents

A method for hydrolysing proteins

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Publication number
AU6564094A
AU6564094A AU65640/94A AU6564094A AU6564094A AU 6564094 A AU6564094 A AU 6564094A AU 65640/94 A AU65640/94 A AU 65640/94A AU 6564094 A AU6564094 A AU 6564094A AU 6564094 A AU6564094 A AU 6564094A
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AU
Australia
Prior art keywords
protein
preparation
proteolytic
hydrolysate
hydrolysis
Prior art date
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Granted
Application number
AU65640/94A
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AU681653B2 (en
Inventor
Gitte Budolfsen
Kim Hansen
Peter Hvass
Per Munk Nielsen
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Novozymes AS
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Novo Nordisk AS
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Publication of AU6564094A publication Critical patent/AU6564094A/en
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Publication of AU681653B2 publication Critical patent/AU681653B2/en
Assigned to NOVOZYMES A/S reassignment NOVOZYMES A/S Alteration of Name(s) in Register under S187 Assignors: NOVO NORDISK A/S
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/01Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01003Triacylglycerol lipase (3.1.1.3)
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23CDAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00Milk preparations; Milk powder or milk powder preparations
    • A23C9/12Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A23C9/1322Inorganic 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
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • A23J3/342Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of collagen; of gelatin
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • A23J3/343Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • A23J3/343Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
    • A23J3/344Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins of casein
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • A23J3/345Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of blood proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/346Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L15/00Egg products; Preparation or treatment thereof
    • A23L15/25Addition or treatment with microorganisms or enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L27/00Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20Synthetic spices, flavouring agents or condiments
    • A23L27/21Synthetic spices, flavouring agents or condiments containing amino acids
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L27/00Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/20Synthetic spices, flavouring agents or condiments
    • A23L27/24Synthetic spices, flavouring agents or condiments prepared by fermentation
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • A23L33/18Peptides; Protein hydrolysates
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/645Proteins of vegetable origin; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/65Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/66Enzymes
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
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    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/48Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
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    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/48Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/58Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi
    • C12N9/62Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from fungi from Aspergillus
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    • C12P21/00Preparation of peptides or proteins
    • C12P21/06Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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    • C12Y304/21004Trypsin (3.4.21.4)
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    • C12Y304/21Serine endopeptidases (3.4.21)
    • C12Y304/21014Microbial serine proteases (3.4.21.14)
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    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61Q5/02Preparations for cleaning the hair

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Description

A METHOD FOR HYDROLYSING PROTEINS
TECHNICAL FIELD
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.
BACKGROUND ART
Methods of hydrolysing proteins have been described in e.g. GB Patent 1,507,380, US Patent 3,723,250, and International Patent Application Nos. WO 89/00272, WO 92/13964, and WO 90/05462.
There is a need for methods of hydrolysing proteins leading to a high degree of protein hydrolysis and to hydrolysates with excellent organoleptic properties.
SUMMARY OF THE INVENTION
It has now been found that a proteolytic enzyme preparation derived from Aspergillus oryzae and supplied from Novo Nordisk A/S under the tradename Flavourzyme™ possesses excellent protein hydrolysing properties, for example is it possible to obtain a high degree of hydrolysis and a non-bitter hydrolysate.
Accordingly, 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 Flavourzyme™.
In its second aspect, the invention provides protein hydrolysates obtained by the method of the invention.
In further aspects, the invention provides food and non-food products comprising a protein hydrolysate of the invention. BRIEF DESCRIPTION OF DRAWINGS
The present invention is further illustrated by reference to the accompanying drawings, in which:
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 ιo method of invention (■: Flavourzyme™, A : Corolase™ 7092, O: Corolase™ 7093, ♦: Alcalase™, D: Neutrase™) ; and
Fig. 3 shows the degree of hydrolysis (%DH) of Na- caseinate after 22 hours of hydrolysis according to the method of invention (■: Flavourzyme™, A : Corolase™ 7092, O: Corolase™ is 7093, ♦: Alcalase™, D: Neutrase™).
DETAILED DISCLOSURE OF THE INVENTION
Characterization of Flavourzyme™ showed that the proteolytic Aspergillus oryzae preparation comprises several proteolytic components. It appeared that the preparation
20 comprises five or more 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.
Accordingly, the present invention provides a method
25 for hydrolysing a protein by incubating with a 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
30100 kD.
In a preferred embodiment, 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
3553 kD, respectively. 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. In the context of this invention, the degree of hydrolysis (DH) is defined by the following formula:
Number of peptide bonds cleaved DH = x 100 %
Total number of peptide bonds
The DH may be calculated according to Adler-Nissen ;
Enzymic Hydrolysis of Food Proteins; Elsvier Applied Science
Publishers Ltd. (1986), p. 122. By using the method of the invention it may be possible to obtain a DH which is higher than 35%, preferably higher than 60%, more preferably higher than 70%, most preferably higher than 80%.
By the method of the invention it may also be possible to obtain a high degree of protein solubility. The degree of protein solubility may be described by way of a
Protein Solubility Index (PSI) as described by Adler-Nissen , op. cit .
In a preferred embodiment, a protein solubility higher than 50% PSI, preferably higher than 70% PSI, more preferably higher than 90% PSI, is obtained.
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.
To obtain a satisfactory degree of hydrolysis, 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. As shown in Example 2, 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.
In accordance with established practice, 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.
In yet another preferred embodiment, incubation of a protein or proteinaceous substrate may be performed with a combination of Flavourzyme™ and one or more other protease preparations. Preferred protease preparations comprises neutral or alkaline proteases. Examples of suitable neutral proteases are neutral proteases derived from Bacillus, preferably from Bacillus εubtilis, such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename Neutrase™. Examples of suitable alkaline proteases are alkaline proteases derived from Bacillus , preferably from Bacillus licheniformis, such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename Alcalase™ 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 Flavourzyme™ and one or more other lipase preparations. Preferred lipase preparations comprises fungal lipases. Examples of 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 Palatase™M; and lipases derived from Aspergillus , preferably from Aspergillus niger such as the enzyme preparation supplied by Novo Nordisk, Denmark, under the tradename Palatase™A.
In another aspect, the present invention provides a protein hydrolysate obtained by the method of the invention.
Determination of AU
The proteolytic activity may be determined with haemoglobin as substrate.
In the Anson-Haemoglobin method for the determination of proteolytic activity denatured haemoglobin is digested, and the undigested haemoglobin is precipitated with trichloroacetic acid (TCA) . The amount of TCA soluble product is determined with phenol reagent, which gives a blue colour with tyrosine and tryptophan. One Anson Unit (AU) is defined as the amount of enzyme which under standard conditions (i.e. 25°C, pH 7.5 and 10 min. reaction time) digests haemoglobin at an initial rate, so that there is liberated per minute an amount of TCA soluble product which gives the same colour with phenol reagent as one milliequivalent of tyrosine.
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.
Determination of LU
Assay for lipase activity :
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.
Food Products
In a further aspect, 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. Thus, 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. For a more detailed description of the general procedure, see applicant's co-pending International Patent Application WO 90/05462) rendered from bone by subjecting crushed bone to the method of the invention. Other proteinaceous by-products from the meat industry may also be used.
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.
Accordingly, 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. Thus, the proteolytic enzyme preparation, e.g. Flavourzyme™, used in the present method or 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. Examples of such 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
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.
Production of very high DH-hydrolysates from gelatine improves gelatine products for incorporation into cosmetics, e.g. creams and shampoos.
The beneficial effect as fermentation medium mentioned 10 above is expected to make the hydrolysates of the invention useful for other fermentations as well.
The following examples further illustrate the present invention, and they are not intended to be in any way limiting to the scope of the invention as claimed.
15 EXAMPLE 1
Beef Hydrolysis
In this example two experiments were carried out using the following enzyme preparations (indicating % w/w based on protein content) : 20 Preparation A) 2% Flavourzyme™ (2.30 AU/g)
Preparation B) 1% Flavourzyme™, 2% Neutrase™ 0.5L, and 0.15% Alcalase™ 2,4L.
All enzymes are available from NOVO NORDISK A/S,
Denmark. Flavourzyme™ is a proteolytic preparation derived from
25.Aspergillus oryzae , Neutrase™ 0.5L is a proteolytic preparation derived from Bacillus subtilis, and Alcalase™ 2.4L is a proteolytic preparation derived from Bacillus licheniformis .
408 g of beef were minced twice and mixed with 392 g of water to a protein content of 10 % w/w. The mixture was
30 blended for 30 sec. and heated to 55°C. Initial osmolality
(mOsm) and soluble dry matter (°Brix) were measured, and the subsequent hydrolysis was monitored by these values.
After four hours of incubation the enzymes were inactivated by heating at 90°C for 30 min. The hydrolysates were subjected to centrifugation at 3000 xG for 15 min. and weighed. The centrifugate was stored in a refrigerator until the next day, and the lipid phase separated off and weighed. pH in centrifugate was 5.85. 5 The results are shown in Tables 1 and 2 below.
Table 1
Initial Osmolality (mOs-m) and Soluble Dry Substance CBrix)
Time Preparation A Preparation B min. mOsm/kg ΔmOsm/kg °Brix mOsm/kg ΔmOsm/kg °Brix
10
0 203 3.9 202 0 4.5
15 360 158 9.9
45 490 287 11.1 484 282 11.1
60 534 331 11.6
15 90 576 373 12.3 534 332 12.3
120 623 420 12.7 579 377 12.8
180 685 482 13.5 624 422 13.3
240 726 523 13.8 663 461 13.7
20 Table 2
Preparation A
Mixture Weight Protein Protein Protein yield DS g % g % %
25 Meat/Water 800 10 80 15
Defatted centrifugate 689 9.46 65.2 81.5 10.6
Dry Substance
3o As calculated from the Tables, the dosages used in these experiments Flavourzyme™ increase yields from usually 45% to 81.5%, and the solubility increases from usually 57% to 89.9% PSI. A degree of hydrolysis 70.2% DH was achieved.
The hydrolysates were served before a tasting panel in a 4% solution. The meat flavour was most pronounced in hydrolysates obtained with Preparation A.
EXAMPLE 2
Hydrolysis at Various pH
In this experiment the method of the invention was performed at pH values in the range of 5-9 with sodium caseinate as substrate.
An 8% (w/w) solution of sodium caseinate (Miprodan™ 30, obtained from MD FOODS Amba, Denmark) was prepared by heating to 80°C. To 800 g of this solution 1 g methyl-4- hydroxybenzoate and 0.15 g propyl-4-hydroxybenzoate were added, and the temperature of the solution was lowered to 50°C. pH was adjusted to the desired value with 4 N NaOH or 4 N HC1.
1% w/w based on the protein content Flavourzyme™ (2.30 AU/g, NOVO NORDISK A/S, Denmark) was added the solution, and the incubation was allowed to proceed to maximal degree of hydrolysis (about 22 hours) .
At pH 9.00, 8.00, and 7.00 the hydrolysis was carried out in a pH stat, and the incubation monitored by consumption of NaOH. At pH 6.00 and 5.00 (initial pH) the hydrolysis was monitored by decreased pH and increased osmolality. Samples were obtained after 0.25, l, 2, 4.5, and 22 hours of incubation. The enzyme was inactivated by heating to 85°C for 3 minutes, followed by cooling in ice-water.
The results are presented in Fig. 1. As evident from this Figure, the process of the invention is capable of exerting hydrolysis in the pH range of from pH 5 to pH 9. EXAMPLE 3
Comparison Example
In this experiment the degree of hydrolysis (DH) obtained by the process of invention (incubation with Flavourzyme™) was compared to the DH obtained by incubation with other known proteolytic preparations.
The experiment was carried out at 50°C as a non-pH stat hydrolysis (no adjustments of pH during hydrolysis) , starting at pH 7.00 adjusted with 4N NaOH. To ensure equal enzyme dosage each hydrolysis was carried out using the same proteolytic activity as expressed in AU/g (cf. above for determination of AU) .
Two protein raw materials were tested: 1) Na-caseinate (supplied by MD Foods Amba, Denmark) 2) Soy protein isolate (supplied by Protein Technologies International, U.S.A.).
2500 g of an 8% (w/w based on protein content) solution of protein raw material was prepared. The protein solution was heat treated at 85°C for 3 minutes and cooled to hydrolysing temperature 50°C.
Samples of 400g were prepared for hydrolysis.
The following enzymes and enzyme dosages were used:
Enzyme Activity, AU/g Dosage per 400g
Flavourzyme, 3.36 0.321 g
Novo Nordisk A/S
Neutrase 0.5L, 0.484 2.231 g
Novo Nordisk A/S
Alcalase 2.4L, 2.58 0.419 g
Novo Nordisk A/S
Corolase 7093, 0.094 11.489 g Rohm GmbH
Corolase 7092, 1.54 1.426 g
Rohm GmbH The hydrolysis was carried on for 22 hours.
The degree of hydrolysis was measured after 5 and 22 hours of hydrolysis, respectively. The results are shown in fig. 2 (soy protein isolate) and fig. 3 (Na-caseinate) . From
5 the results it is evident that hydrolysis by using Flavourzyme clearly results in the highest degree of hydrolysis.
EXAMPLE 4
, Cheese Flavour Production
Whole milk was heat treated for 2 min at 100°C and o subsequently cooled to 50°C. Four portions of 200 ml each were prepared (indicating % w/w based on protein content):
1) 1% Flavourzyme™ (2.30 AU/g)
2) 1% Flavourzyme™ (2.30 AU/g) + 0.15% Palatase™ M 200L (200 LU/g from NOVO NORDISK A/S, Denmark) 5 3) 0.15% Palatase™ M 200L (200 LU/g from NOVO
NORDISK A/S, Denmark) 4) Control
The hydrolysis was carried out for 20 hours at 50°C. Intensive cheese flavour developed in samples l, 2, and 3, 0 while the control was almost neutral.
The flavour-samples 1, 2, and 3, were now diluted in cream cheese, and a fourth sample was prepared as a control. 1.1) 0.5 g of sample No. 1 + 20 g of cheese 2.1) 6.0 g of sample No. 2 + 20 g of cheese 5 3.1) 6.0 g of sample No. 3 + 20 g of cheese
4.1) 6.0 g of fresh milk + 20 g of cheese. Sample No. 1.1 (Flavourzyme™) appeared to have a very strong and intense flavour and a taste of well ripened cheese. Sample No. 2.1 (Flavourzyme™ + Palatase™) appeared to have a 0 well balanced cheese flavour. Compared to sample No. 1.1 and No. 2.1, sample No. 3.1 turned out to give the weakest taste impression. The reference, sample No. 4.1 was evaluated being sour/sourish.
In conclusion it is possible to add the enzymes 5 Flavourzyme™ and the combination of Flavourzyme™ and Palatase™ directly into the milk to form excellent cheese flavour. The traditional way of producing cheese flavour is carried out by adding lipases and proteases to cheese. That is, the use of Flavourzyme™ and Flavourzyme™ in combination with
Palatase™ creates a new and much more simple way of producing cheese flavour.
EXAMPLE 5
Accelerated Yoghurt Fermentation
Whole milk was heat treated for 2 min at 100°C and subsequently cooled to 50°C. Two portions of 200 ml were prepared (indicating % w/w based on protein content) :
1) 1% Flavourzyme™ (2.30 AU/g)
2) Control
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.
1.1) 200 ml of milk + 10 ml of sample No. 1 2.1) 200 ml of milk + 10 ml of sample No. 2 3.1) 200 ml of milk
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 106 bacteria per g.
The fermentation course was followed by measuring pH, cf. Table 4, below.
Table 4
Acidification Course
(pH) 1) Flavour- 2) Blind 3) Blind/Blind zyme™
Initial pH value 6.6 6.5 6.5 pH after 200 min 5.9 6.4 6.4 pH after 360 min 4.4 5.9 6.3
It appears from the pH measurements that the addition of Flavourzyme™ hydrolysed milk accelerates the fermentation, meaning that the production time of fermented milk products can be reduced considerably.

Claims (21)

1. A method for hydrolysing a protein by incubating with a proteolytic enzyme preparation, characterized in that the proteolytic preparation is derived from Aspergillus oryzae and comprises at least five proteolytic components each 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 100 kD.
2. The method according to claim 1 in which the proteolytic preparation comprises at least five proteolytic components having the approximate molecular weights 23 kD, 31 kD, 35 kD, 38 kD and 53 kD, respectively.
3. The method according to claim 1 or 2, by which there is obtained a degree of protein hydrolysis (DH) higher than 35%, preferably higher than 60%, more preferably higher than 70%, especially higher than 80%.
4. The method according to claim 1, by which there is obtained a degree of protein solubility (Protein Solubility Index (PSI)) higher than 50%PSI, preferably higher than 70% PSI, more preferably higher than 90%PSI.
5. The method according to any of claims 1-4, in which the protein is a vegetable protein, preferably 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.
6. The method according to any of claims 1-4, in which the protein is an animal protein, preferably milk protein, whey protein, casein, meat protein, fish protein, blood protein, egg white, or gelatin.
7. The method according to any of claims 1-6, in which the incubation is performed at a pH of between about 4 and about 10, preferably between about 5 and about 9.
8. The method according to any of claims 1-7, in which the protein is incubated with the proteolytic preparation in combination with one or more other protease preparations.
9. The method according to claim 8, in which the protease preparation is a neutral protease derived from Bacillus , preferably from Bacillus subtilis .
10. The method according to claim 8, in which the protease preparation is an alkaline protease derived from Bacillus , preferably from Bacillus lichenifor is .
11. The method according to any of the claims 1-10, in which the protein is incubated with the proteolytic preparation in combination with one or more lipase preparations.
12. The method according to claim 11, in which the lipase preparation comprises a fungal lipase derived from Mucor, preferably from Rhizomucor miehei .
13. The method according to claim 11, in which the lipase preparation comprises a fungal lipase derived from Aspergillus , preferably from Aspergillus niger.
14. The method according to any of the claims 1-13, wherein the protein is hydrolysed and simultaneously fermented to a food product.
15. A protein hydrolysate obtained by the method according to any of the claims 1-13.
16. The protein hydrolysate according to claim 15, wherein the hydrolysed protein is an animal protein selected from the group consisting of milk protein, whey protein, casein, meat proteins, fish protein, blood protein, egg white and gelatin.
17. The protein hydrolysate according to claim 15, wherein the hydrolysed protein is a vegetable protein selected from the group consisting of soy protein; grain proteins such as wheat gluten or zein; rape seed protein; alfalfa protein; pea protein; fabaceous bean protein; cotton seed protein; and sesame seed protein.
18. A food product comprising a protein hydrolysate according to any of the claims 15-17.
19. The food product according to claim 18, being an ingredient of mother milk substitute; a cheese flavour product; an enzymatically produced HVP; a protein enriched dietetic product; a soup-, bouillon- or meat-flavour product; a meat extract product; or a hydrolysate for improving the production of a starter culture.
20. A non-food product comprising a protein hydroly¬ sate according to any of the claims 15-17.
21. The non-food product according to claim 20, being a pet food, a cosmetic, or a fermentation broth.
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Families Citing this family (69)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2213164T3 (en) * 1994-10-26 2004-08-16 Novozymes A/S PRODUCTION METHOD OF A MILK PROTEIN HYDROLYZE.
DE19632455C1 (en) * 1996-08-12 1997-08-21 Cpc Maizena Gmbh Production of liquid protein hydrolysate
BR9712611A (en) * 1996-10-30 1999-10-26 Novo Nordisk As Process for producing in food flavoring agent.
AU720774B2 (en) * 1996-10-30 2000-06-08 Novozymes A/S Method of producing food flavoring agent
ES2116937B1 (en) * 1996-11-29 1999-03-01 Consejo Superior Investigacion PROCEDURE FOR OBTAINING VEGETABLE PEPTONES OF HIGH DEGREE OF HYDROLYSIS AND ITS APPLICATIONS.
JP2001507223A (en) * 1996-12-23 2001-06-05 デーエスエム ナムローゼ フェンノートシャップ Flavor enhancer
US6007851A (en) * 1996-12-23 1999-12-28 Gist-Brocades, B.V. Process for producing a flavor enhancer
WO1998027827A1 (en) * 1996-12-23 1998-07-02 Dsm N.V. Method for producing a protein hydrolysate
ATE414775T1 (en) 1997-05-16 2008-12-15 Novozymes Inc POLYPEPTIDES WITH PROLYLDIPEPTIDYLAMINOPEPTIDASE ACTIVITY AND NUCLEIC ACIDS CODING THEREFOR
EP0984703B1 (en) 1997-05-16 2003-04-09 Novozymes Biotech, Inc. Methods of producing protein hydrolysates
JP2003511093A (en) 1999-10-20 2003-03-25 ノルデュール・イーエイチエフ Protein hydrolyzate produced using marine protease
FR2804691B1 (en) 2000-02-04 2003-11-07 Roquette Freres NITROGEN COMPOSITION RESULTING FROM HYDROLYSIS OF CORN GLUTEN AND METHOD FOR PRODUCING THE SAME
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 (en) * 2000-10-02 2002-04-12 김강권 Process for preparation of hypoallergenic casein protein hydrolysate and hypoallergenic infant formula containing casein protein hydrolysate
WO2002032231A1 (en) 2000-10-19 2002-04-25 Edens, Luppo Protein hydrolysates
WO2002069732A1 (en) * 2001-03-05 2002-09-12 Council Of Scientific And Industrial Research Process for preparation of protein hydrolysate from soy flour
CN1225185C (en) * 2001-03-05 2005-11-02 科学和工业研究委员会 Process for preparation of protein hydrolysate from milk protein
KR20030028089A (en) * 2001-09-27 2003-04-08 주식회사 태훈바이오 Reaction flavors and method of preparing the same from snow crab by-products
US7404977B2 (en) 2001-12-21 2008-07-29 Dsm Ip Assets B.V. Rennets
FR2836014B1 (en) * 2002-02-15 2004-07-23 Gervais Danone Sa NEW PROCESS FOR THE MANUFACTURE OF FERMENTATED DAIRY PRODUCTS USING BACTERIAL ENZYMES
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
WO2004098309A1 (en) * 2003-05-05 2004-11-18 Unilever N.V. Hydrolysed casein product comprising tripeptides ipp and/ or vpp
ITMI20031528A1 (en) * 2003-07-25 2005-01-26 Ct Sperimentale Del Latte S P A FERMENTATION SOIL BASED ON A HIGHLY HYDROLISED PROTEIN SUBSTRATE
DE602004017202D1 (en) * 2003-08-01 2008-11-27 Calpis Co Ltd CASEIN HYDROLYZATE, METHOD FOR THE PRODUCTION THEREOF AND THE USE THEREOF
US7485323B2 (en) 2005-05-31 2009-02-03 Gelita Ag Process for making a low molecular weight gelatine hydrolysate and gelatine hydrolysate compositions
WO2007066841A1 (en) * 2005-12-09 2007-06-14 Serombio Co., Ltd. Yeast hydrolysate containing cyclo-his-pro and method for producing the same
JP4726129B2 (en) * 2006-01-17 2011-07-20 学校法人北里研究所 Bifidobacterium growth-promoting peptide
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
US7998476B2 (en) 2006-11-22 2011-08-16 Standard Biologics, Inc. Method of treatment using Aspergillus oryzae protease
EP1969950A1 (en) * 2007-03-12 2008-09-17 Cargill, Incorporated Partially hydrolysed cereal protein
FR2925325B1 (en) * 2007-12-21 2010-02-26 Vincience USE OF A COLZA HYDROLYZATE AS AN ACTIVATOR ACTIVE INGREDIENT OF AQUAPORIN SYNTHESIS
ES2378285T3 (en) * 2008-06-03 2012-04-10 Novozymes A/S Method for the production of a wheat protein hydrolyzate
DK2300606T3 (en) * 2008-06-03 2012-04-16 Novozymes As Process for the preparation of a casein hydrolyzate
WO2010009400A1 (en) * 2008-07-18 2010-01-21 Novozymes A/S Method for preparing a protein hydrolysate
US8633030B2 (en) 2008-12-16 2014-01-21 Novozymes, Inc. Polypeptides having carboxypeptidase activity and polynucleotides encoding same
CN101518285B (en) * 2009-03-30 2011-07-20 山东轻工业学院 Method for producing cheese
EP2258208A1 (en) * 2009-06-02 2010-12-08 University of Limerick Protein Product With Modified Antigenicity
JP2013516432A (en) * 2009-12-30 2013-05-13 ソレイ リミテッド ライアビリティ カンパニー Method for recovering Kunitz trypsin inhibitor protein from a soybean processing stream
NO20100370A1 (en) 2010-03-08 2011-09-09 Marine Bioproducts As Peptide material, feed compositions and preparations, and uses thereof.
JP5952826B2 (en) * 2010-12-06 2016-07-13 カーギル インコーポレイテッド Cereal protein liquefaction method
CN102550895A (en) * 2010-12-28 2012-07-11 雀巢公司 Enzyme preparation from solid fermentation of food
EP2489281A1 (en) * 2011-02-17 2012-08-22 University of Limerick A casein hydrolysate
WO2012121612A1 (en) * 2011-03-04 2012-09-13 Meat Biologics Research Limited Method of manufacture of an edible composition
WO2012121611A2 (en) * 2011-03-04 2012-09-13 Meat Biologics Research Limited Edible composition and method of manufacture
UA111078C2 (en) * 2011-05-03 2016-03-25 Нестек С.А. Hydrolysate of a protein substrate and a method for obtaining thereof
ES2395982B1 (en) * 2011-07-08 2014-01-22 Apc Europe S.A. Food supplement, method to increase the bacterial mass in the rumen of a ruminant and food preparation and corresponding uses.
AU2012320643B2 (en) * 2011-10-03 2015-12-10 Specialites Pet Food Animal digests having enhanced palatability for use in pet food
CA2937613C (en) 2013-01-23 2021-03-02 Bottled Science Limited Skin enhancing beverage composition
US9139458B2 (en) 2013-03-15 2015-09-22 Janet Angel Compositions and methods of use
JP6207922B2 (en) * 2013-08-14 2017-10-04 マルハニチロ株式会社 Lever stab food that can be stored at room temperature for a long time
CN103549435A (en) * 2013-11-19 2014-02-05 哈尔滨艾克尔食品科技有限公司 Method for preparing oral whey protein peptide liquid
JP6481923B2 (en) * 2014-05-02 2019-03-13 奥野製薬工業株式会社 Livestock meat improver
US20170150737A1 (en) * 2014-05-28 2017-06-01 Hajime Hatta Highly emulsifiable albumen hydrolysate
WO2015181675A1 (en) * 2014-05-29 2015-12-03 Lb Lyopharm S.R.L. A process for the preparation of functional plant proteins
US10619177B2 (en) 2014-12-01 2020-04-14 Novozymes A/S Method for producing a protein hydrolysate
WO2016156077A1 (en) 2015-03-30 2016-10-06 Nestec S.A. Milk-based protein hydrolysates and compositions made thereof
CN104719920B (en) * 2015-04-09 2017-03-08 广州超亚化妆品科技有限公司 Polypeptide amino acid mixture and preparation method and application
CN105077158B (en) * 2015-08-24 2018-05-01 王婧婧 A kind of preparation method of cheese's flavor frankincense base
DE102017200434A1 (en) 2016-01-13 2017-07-13 Robert Finke Whey-containing body care product
CN107772077A (en) * 2016-08-30 2018-03-09 中粮营养健康研究院有限公司 A kind of method of the protein solubility of rape cake and raising rape cake
CN107788223A (en) * 2016-08-30 2018-03-13 中粮营养健康研究院有限公司 A kind of method for the protein solubility for improving rape cake
EP3512359A4 (en) * 2016-09-14 2020-04-01 Glanbia Nutritionals (Ireland) Ltd. Agglomerated protein products and method for making
CN106616995A (en) * 2016-12-27 2017-05-10 济南邦地生物工程有限公司 Brain-strengthening oral liquid and production method thereof
WO2019090011A1 (en) * 2017-11-03 2019-05-09 Cargill, Incorporated Pea protein hydrolysate
CN108771194A (en) * 2018-07-02 2018-11-09 广东海洋大学 A method of assist enzymolysis aquatic product protein to prepare seafood seasoning packet using high temperature resistant bacillus licheniformis
CN110810687B (en) * 2019-11-21 2022-08-23 江南大学 Method for preparing oligopeptide refreshing beverage by hydrolyzing sesame protein by endogenous endopeptidase and exopeptidase
EP4289277A1 (en) * 2021-02-04 2023-12-13 Amano Enzyme U.S.A. Co., Ltd. Natural cheese production method

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3306009C2 (en) * 1983-02-22 1994-02-17 Roehm Gmbh Process for the preparation of protein hydrolyzates
CN86108557A (en) * 1986-12-20 1988-07-13 同济大学 With proteinaceous waste material is material, enzyme method method of extracting protein hydrolystate and products thereof
CH679542A5 (en) * 1989-11-27 1992-03-13 Nestle Sa

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