WO2021089243A1 - Verfahren zur herstellung von kollagenpeptiden aus knochen, und hergestellte kollagenpeptide - Google Patents
Verfahren zur herstellung von kollagenpeptiden aus knochen, und hergestellte kollagenpeptide Download PDFInfo
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- WO2021089243A1 WO2021089243A1 PCT/EP2020/077092 EP2020077092W WO2021089243A1 WO 2021089243 A1 WO2021089243 A1 WO 2021089243A1 EP 2020077092 W EP2020077092 W EP 2020077092W WO 2021089243 A1 WO2021089243 A1 WO 2021089243A1
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- bones
- collagen peptides
- collagen
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 82
- 108010035532 Collagen Proteins 0.000 title claims abstract description 82
- 229920001436 collagen Polymers 0.000 title claims abstract description 82
- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 78
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 69
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 68
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 45
- 108091005804 Peptidases Proteins 0.000 claims abstract description 20
- 239000004365 Protease Substances 0.000 claims abstract description 20
- 102000035195 Peptidases Human genes 0.000 claims abstract description 18
- 239000007900 aqueous suspension Substances 0.000 claims abstract description 18
- 239000007864 aqueous solution Substances 0.000 claims abstract description 15
- 238000002803 maceration Methods 0.000 claims abstract description 14
- 238000010438 heat treatment Methods 0.000 claims abstract description 12
- 239000000725 suspension Substances 0.000 claims abstract description 11
- 239000002245 particle Substances 0.000 claims abstract description 7
- 241000251539 Vertebrata <Metazoa> Species 0.000 claims abstract description 4
- 239000002253 acid Substances 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 241000283690 Bos taurus Species 0.000 claims description 4
- 238000001238 wet grinding Methods 0.000 claims description 4
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 claims description 3
- 102000004882 Lipase Human genes 0.000 claims description 3
- 108090001060 Lipase Proteins 0.000 claims description 3
- 239000004367 Lipase Substances 0.000 claims description 3
- 238000009835 boiling Methods 0.000 claims description 3
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 claims description 3
- 238000006911 enzymatic reaction Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 229960002591 hydroxyproline Drugs 0.000 claims description 3
- 235000019421 lipase Nutrition 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 claims description 3
- 244000063299 Bacillus subtilis Species 0.000 claims description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 2
- 241000124008 Mammalia Species 0.000 claims description 2
- 101710118538 Protease Proteins 0.000 claims description 2
- 108010022999 Serine Proteases Proteins 0.000 claims description 2
- 102000012479 Serine Proteases Human genes 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 claims description 2
- 238000009837 dry grinding Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000005374 membrane filtration Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 229920000159 gelatin Polymers 0.000 description 23
- 235000019322 gelatine Nutrition 0.000 description 23
- 108010010803 Gelatin Proteins 0.000 description 15
- 235000011852 gelatine desserts Nutrition 0.000 description 15
- 239000008273 gelatin Substances 0.000 description 14
- 239000001828 Gelatine Substances 0.000 description 8
- 230000007071 enzymatic hydrolysis Effects 0.000 description 8
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 8
- 239000008346 aqueous phase Substances 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000009997 thermal pre-treatment Methods 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 108090000787 Subtilisin Proteins 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- 238000001155 isoelectric focusing Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 102000012422 Collagen Type I Human genes 0.000 description 1
- 108010022452 Collagen Type I Proteins 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 241001620634 Roger Species 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 229910001576 calcium mineral Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 238000005115 demineralization Methods 0.000 description 1
- 230000002328 demineralizing effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 238000013021 overheating Methods 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
-
- 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
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
-
- 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
- A23J3/04—Animal proteins
- A23J3/06—Gelatine
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- 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)
- C12Y304/21062—Subtilisin (3.4.21.62)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- the present invention relates to a method for producing collagen peptides from bones.
- the invention also relates to collagen peptides which are produced according to this method.
- Collagen peptides are formed during the hydrolysis of the animal structural protein collagen, in particular through enzymatic hydrolysis. Alternative names are therefore collagen hydrolyzate or hydrolyzed collagen. In the case of animal bones as the starting material, it is particularly type I collagen.
- Collagen peptides are used in a variety of ways, especially in the food industry, on the one hand because of their physiological effect in food supplements or so-called "functional food", but also from the point of view of food technology such as, for example, as an emulsifier, stabilizer, binder, etc.
- a characteristic property of collagen peptides is their solubility in cold water and their inability to form a gel. This allows collagen peptides to be differentiated from gelatin, which is denatured and only slightly hydrolyzed collagen.
- Collagen peptides have a molecular weight of less than 25,000 Da, mostly even less than 10,000 Da, while the molecular weights of gelatine are significantly higher.
- Collagen peptides are normally made from gelatin as an intermediate product (see, for example, R. Schrieber and H. Gareis: Gelatin Handbook, 2007, chapter 2.2.11).
- gelatine is again produced from bones in a multistage process, the essential steps of which are demineralization of the bones in a strongly acidic environment (Maceration) and a subsequent treatment in a strongly alkaline environment (liming) in order to then be able to extract the gelatine at an elevated temperature (typically between 50 and 100 ° C) in several stages (see Gelatin Handbook, Chapter 2.2.5 ).
- the crushed bones are treated with dilute hydrochloric acid in a countercurrent process over a period of about one week in order to remove the mineral components (calcium carbonate and calcium phosphate) from the bone tissue (see Gelatin Handbook, Chapter 2.2.1.1).
- the product obtained by this process is called ossein.
- a relevant cost factor in maceration is the required cooling due to the exothermic reaction of the hydrochloric acid with the calcium minerals.
- Another disadvantage is the high chloride load in the wastewater.
- the subsequent liming of the ossification is necessary to enable the gelatine to be extracted effectively.
- the typical liming process involves treatment with a calcium hydroxide suspension (pH value above 12) over a period of several months (see Gelatin Handbook, Chapter 2.2.4.1).
- the use of stronger alkalis can shorten the treatment time (e.g. to a few days when using sodium hydroxide), but this leads to a loss of yield.
- type B bone gelatin which is characterized by an isoelectric point (IEP) of less than 5.6, typically in the range of 4.8 to 5.5.
- the IEP corresponds to that pH value at which the polypeptide chains of the gelatin (or the collagen peptides produced from it) have a neutral overall charge.
- the relatively low IEP of type B gelatin results from the fact that the amino acids asparagine and glutamine are almost completely converted to aspartic acid and glutamic acid during liming.
- Type B gelatine has a significantly higher viscosity than type A gelatine with the same gel strength and is therefore preferred for most fields of application.
- Type A gelatine in which the osseous is extracted in an acidic environment without liming, therefore only plays a subordinate role.
- Type A gelatins have an IEP of over 6, in the case of type A bone gelatin typically in the range between 6 and 8 (in the case of pork skin gelatin in the range 8 to 9).
- the invention is based on the object of proposing an alternative method for the production of collagen peptides, in which the disadvantages of the method described above using bone gelatin of type B as an intermediate product can be entirely or partially avoided.
- this object is achieved according to the invention in that it comprises the following steps: a) providing bones from vertebrates; b) mechanical comminution of the bones down to a particle size of less than 1,000 ⁇ m, preferably less than 500 ⁇ m, more preferably less than 300 ⁇ m, at a temperature of less than 70 ° C.
- step c) heating the comminuted bones in an aqueous suspension to a temperature of over 100 ° C., preferably over 120 ° C., more preferably over 130 ° C., for a period of 1 to 30 min, preferably 2 to 10 min, more preferably 4 to 8 minutes; d) adding one or more proteases to the suspension in order to obtain an aqueous solution of collagen peptides; and e) separating the aqueous solution of collagen peptides from the crushed bones, the method comprising neither maceration of the bones with an acid nor liming of the bones with a base, and wherein the boiling provided in step a) does not include maceration and liming were subjected.
- collagen peptides can be produced by direct enzymatic treatment of the bone material by means of proteases, without the detour via the production of bone gelatin.
- the method according to the invention therefore explicitly dispenses with maceration and / or liming of the bones, which drastically shortens the entire duration of the process until the collagen peptides are obtained: while maceration and liming can take several months in extreme cases, but at least several days the inventive method can be carried out within a few hours.
- the energy requirement and the wastewater pollution are significantly reduced in the method according to the invention compared to the prior art.
- the term "maceration” refers to a treatment with an acid at a pH of less than 1
- the term “liming” refers to a treatment with a base at a pH of over 12 Roger that.
- bones from any vertebrate animal can be used as the starting material for the method according to the invention, e.g. also from birds or fish.
- the method is preferably carried out with bones from mammals, in particular with bones from cattle.
- the cleaning of the bones preferably includes a treatment with one or more enzymes, preferably with proteases and / or lipases. While the lipases are used for defatting, non-collagenous proteins can be broken down and removed by means of proteases. A hydrolysis of the collagen through the proteases do not occur to any significant extent before the bones are comminuted.
- the bones Before being comminuted, the bones advantageously have a fat content of less than 4% by weight, preferably less than 1% by weight, more preferably less than 0.5% by weight.
- the bones have a collagen content in relation to the total protein content of at least 55%, preferably from 70 to 90%, before comminution.
- the collagen content is determined using the hydroxyproline content multiplied by the factor 7.3, and the total protein content is determined using the Kjeldahl nitrogen content multiplied by the factor 6.25.
- the factors mentioned take into account the proportion of hydroxyproline and nitrogen in collagen, which differs from the corresponding proportions in the total protein.
- the mechanical comminution of the preferably cleaned bones down to a particle size of less than 1,000 ⁇ m is an essential feature of the method according to the invention. Due to the small particle size, direct enzymatic hydrolysis of the collagen in the bone material is made possible without the need for the pretreatment procedures known from the prior art, such as maceration or liming.
- the mechanical comminution can include dry milling or wet milling of the bones, with wet milling in an aqueous suspension being preferred. During the shredding, the temperature is kept below 70 ° C in order to avoid local overheating of the material.
- the minced bones are heated in an aqueous suspension to a temperature of over 100 ° C., a maximum of 30 minutes being sufficient for this thermal pretreatment.
- the collagen is denatured and made accessible for enzymatic hydrolysis.
- the weight fraction of crushed Bone in the aqueous suspension is preferably from 0.05 to 0.5 kg / l, preferably from 0.1 to 0.3 kg / l, more preferably from 0.15 to 0.2 kg / l.
- the thermal pretreatment of the comminuted bones can be further accelerated and / or intensified by an additional energy input by means of cavitation, e.g. by ultrasound or a high-pressure homogenizer.
- cavitation e.g. by ultrasound or a high-pressure homogenizer.
- Another possibility is to apply alternating electric fields to the suspension.
- Another advantage of the method according to the invention is that the isoelectric point of the collagen peptides produced can be influenced in a simple manner by setting a corresponding pH value in the aqueous suspension while the comminuted bones are being heated.
- collagen peptides with a high or low IEP may be preferred, whereby the differences in the properties are less serious here than with gelatine of type A or type B.
- the pH of the aqueous suspension is adjusted to a range from 5 to 7, preferably from 6 to 7, before heating. if the pH value is not adjusted.
- the pH of the aqueous suspension is adjusted to a range from 7 to 9, preferably from 7.9 to 8.6, before heating.
- the aqueous suspension is cooled to a temperature in the range from 40 to 60 ° C. before the one or more proteases are added.
- the activity optima of the proteases, which are typically used for the enzymatic hydrolysis of the collagen, are in this temperature range.
- the one or more proteases which are added to the aqueous suspension after heating are favorably selected from microbial endoproteases, preferably serine proteases, in particular from Bacillus subtilis.
- the use of such enzymes for the hydrolysis of collagen is known from the prior art.
- a protease that is frequently used is, for example, subtilisin.
- the one or more proteases are typically added in an amount of from 0.01 to 0.5% by weight, based on the dry mass of the comminuted bones, preferably from 0.02 to 0.2% by weight, more preferably from 0 .03 to 0.1% by weight.
- the enzymatic reaction is preferably carried out for a period of from 0.5 to 4 hours, more preferably from 1 to 3 hours.
- the minced bones are subjected once more to steps c) to e) after the aqueous solution of collagen peptides has been separated off.
- steps c) to e) after the aqueous solution of collagen peptides has been separated off.
- the separation of the aqueous solution of collagen peptides from the pulverized bones preferably comprises a filtration, in particular a membrane filtration. This means that even the smallest particles of the crushed bones and other solids can be removed.
- the aqueous solution of collagen peptides can preferably be subjected to an ion exchange, in particular desalination.
- the method according to the invention further comprises drying the aqueous solution of collagen peptides in order to obtain a powder of collagen peptides, in particular by means of Spray drying.
- the aqueous solution can be concentrated beforehand with the aid of evaporators.
- the present invention also relates to collagen peptides which are produced by the method according to the invention.
- the collagen peptides according to the invention typically have a weight average molecular weight of less than 25,000 Da, preferably less than 10,000 Da, more preferably less than 5,000 Da.
- the collagen peptides have a high isoelectric point of over 5.6, in particular of over 6.0.
- the collagen peptides have a low isoelectric point of below 5.6, in particular from 5.2 to 5.6.
- Fig. 1 Charge distributions of collagen peptides according to the prior art
- Example 1 Production of collagen peptides from bone on a laboratory scale
- Cattle bones are pre-cleaned using hot water and defleading and degreasing supported by proteases and then ground to a bone powder with a particle size distribution of d50 ⁇ 350 ⁇ m and d90 ⁇ 700 ⁇ m.
- the bone powder is then mixed with the same mass of water and heated in a microwave oven with stirring for approx. 1 min at 120 to 130 ° C. After cooling (approx. 20 min) to below 100 ° C, 0.1% by weight (based on the bone powder mass) of the protease subtilisin is added and the suspension is stirred at 60 ° C. Due to the enzymatic reaction with the formation of soluble collagen peptides, the concentration of the aqueous phase increases over time, as indicated in Table 1. The concentration is measured with a refractometer which is calibrated to the unit ° Brix for measuring sucrose.
- the supernatant is filtered and desalted.
- the collagen peptides are concentrated and dried.
- the IEP of the collagen peptides is 6.23.
- the distribution of the molecular charges of these collagen peptides according to the invention can be determined by means of isoelectric focusing. A corresponding chromatogram is shown in FIG. 1, the pH value in the gel being indicated on the left and the three traces being covered as follows:
- Lane No. 2 Bone gelatin collagen peptides of type B according to the prior art (with maceration and liming)
- Lane no. 3 collagen peptides according to the invention according to the above example
- the charges of the molecules are in principle similar in both samples, the collagen peptides according to the invention in this case also showing bands of negative, ie alkaline, molecules.
- the pH value during denaturation determines the position of the bands and thus the isoelectric point of the collagen peptides.
- Example 2 Production of collagen peptides from bone on a pilot scale
- a 15% strength by weight aqueous suspension of cleaned bone powder (d90 ⁇ 700 ⁇ m) from cattle bones is placed in a stirred container, the bones having been cleaned beforehand as in Example 1.
- the ratio of total protein to collagen is 1.7 (normalized to dry matter minus fat content).
- the pH of the suspension is adjusted to 6.5.
- the suspension is pumped through a heat exchanger and heated to 130 ° C. This temperature is held for about 6 minutes.
- the suspension is then cooled to approx. 60 ° C. using a heat exchanger and collected in a stirred tank. 0.05% by weight (based on the dry bone mass) of the protease subtilisin are added.
- the enzymatic hydrolysis is terminated by heating the suspension to 85 ° C. for 5 minutes.
- the aqueous phase is separated off with a decanter centrifuge and collected in a container.
- the solid phase is treated again in the same way as described above for the bone powder (preparation of a suspension, thermal pretreatment, cooling, enzymatic hydrolysis and separation of the aqueous phase by decanting).
- aqueous phases (collagen peptide solutions) from both runs are combined and, for further purification, filtered, desalted, concentrated and dried using suitable methods.
- the yield of this process is approx. 16 to 19% by weight of collagen peptides, based on the bone mass used.
- the quality of the collagen peptides can be e.g. B. on the basis of high transmission values of aqueous solutions with a concentration of 20 wt.% At the wavelengths 450 nm and 620 nm can be evaluated.
- the measured values and the respective quality standard are given in table 2:
- the weight average molecular weight of the collagen peptides according to the example 2 is in the range of 3,000 ⁇ 500 Da.
- the molecular weight distribution of the collagen peptides according to the invention can be influenced by the choice of enzyme, the amount of enzyme and the reaction time.
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Abstract
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Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2020378186A AU2020378186A1 (en) | 2019-11-08 | 2020-09-28 | Method for producing collagen peptides from bones, and produced collagen peptides |
BR112022008115A BR112022008115A2 (pt) | 2019-11-08 | 2020-09-28 | Processo para a produção de peptídeos de colágeno a partir de ossos e peptídeos de colágeno produzidos |
EP20785702.0A EP4017272A1 (de) | 2019-11-08 | 2020-09-28 | Verfahren zur herstellung von kollagenpeptiden aus knochen, und hergestellte kollagenpeptide |
JP2022526475A JP2023502010A (ja) | 2019-11-08 | 2020-09-28 | 骨からコラーゲンペプチドを生成するための方法、及び生成されたコラーゲンペプチド |
MX2022005337A MX2022005337A (es) | 2019-11-08 | 2020-09-28 | Metodo para producir peptidos de colageno a partir de huesos y peptidos de colageno producidos. |
CN202080077345.0A CN114845560A (zh) | 2019-11-08 | 2020-09-28 | 从骨骼生产胶原蛋白肽的方法和生产的胶原蛋白肽 |
US17/736,430 US20220256886A1 (en) | 2019-11-08 | 2022-05-04 | Method for producing collagen peptides from bones, and produced collagen peptides |
ZA2022/05099A ZA202205099B (en) | 2019-11-08 | 2022-05-09 | Method for producing collagen peptides from bones, and produced collagen peptides |
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DE102019130196.3A DE102019130196A1 (de) | 2019-11-08 | 2019-11-08 | Verfahren zur Herstellung von Kollagenpeptiden aus Knochen, und hergestellte Kollagenpeptide |
DE102019130196.3 | 2019-11-08 |
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US17/736,430 Continuation US20220256886A1 (en) | 2019-11-08 | 2022-05-04 | Method for producing collagen peptides from bones, and produced collagen peptides |
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US (1) | US20220256886A1 (de) |
EP (1) | EP4017272A1 (de) |
JP (1) | JP2023502010A (de) |
CN (1) | CN114845560A (de) |
AU (1) | AU2020378186A1 (de) |
BR (1) | BR112022008115A2 (de) |
DE (1) | DE102019130196A1 (de) |
MX (1) | MX2022005337A (de) |
WO (1) | WO2021089243A1 (de) |
ZA (1) | ZA202205099B (de) |
Citations (5)
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US4176199A (en) * | 1978-05-15 | 1979-11-27 | Sugardale Foods, Incorporated | Extraction of protein from edible beef bones and product |
WO2006121361A1 (en) * | 2005-05-12 | 2006-11-16 | Halina Grabek | Process for the preparation of gelatine and gelatine hydrolyzate |
CN101787078A (zh) * | 2009-01-23 | 2010-07-28 | 香港百特有限公司 | 一种胶原蛋白多肽及其制备方法和用途 |
DE102012110612A1 (de) * | 2012-11-06 | 2014-05-08 | Gelita Ag | Kollagenhydrolysat und dessen Verwendung |
CN103937859A (zh) * | 2013-01-21 | 2014-07-23 | 中国科学院理化技术研究所 | 一种以骨粉为原料由酶法制取明胶的方法 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN1196758C (zh) * | 2002-08-08 | 2005-04-13 | 中国科学院理化技术研究所 | 酶降解骨胶原制备明胶的方法 |
CN103060406B (zh) * | 2011-10-18 | 2015-02-18 | 中国科学院理化技术研究所 | 从牲畜骨中提取低分子量胶原蛋白的方法 |
CN109735591B (zh) * | 2019-03-14 | 2020-11-10 | 舜甫科技(固安)有限公司 | 一种牛骨胶原蛋白肽及其生产方法 |
-
2019
- 2019-11-08 DE DE102019130196.3A patent/DE102019130196A1/de active Pending
-
2020
- 2020-09-28 WO PCT/EP2020/077092 patent/WO2021089243A1/de active Application Filing
- 2020-09-28 JP JP2022526475A patent/JP2023502010A/ja active Pending
- 2020-09-28 EP EP20785702.0A patent/EP4017272A1/de active Pending
- 2020-09-28 MX MX2022005337A patent/MX2022005337A/es unknown
- 2020-09-28 AU AU2020378186A patent/AU2020378186A1/en active Pending
- 2020-09-28 CN CN202080077345.0A patent/CN114845560A/zh active Pending
- 2020-09-28 BR BR112022008115A patent/BR112022008115A2/pt unknown
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2022
- 2022-05-04 US US17/736,430 patent/US20220256886A1/en active Pending
- 2022-05-09 ZA ZA2022/05099A patent/ZA202205099B/en unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4176199A (en) * | 1978-05-15 | 1979-11-27 | Sugardale Foods, Incorporated | Extraction of protein from edible beef bones and product |
WO2006121361A1 (en) * | 2005-05-12 | 2006-11-16 | Halina Grabek | Process for the preparation of gelatine and gelatine hydrolyzate |
CN101787078A (zh) * | 2009-01-23 | 2010-07-28 | 香港百特有限公司 | 一种胶原蛋白多肽及其制备方法和用途 |
DE102012110612A1 (de) * | 2012-11-06 | 2014-05-08 | Gelita Ag | Kollagenhydrolysat und dessen Verwendung |
CN103937859A (zh) * | 2013-01-21 | 2014-07-23 | 中国科学院理化技术研究所 | 一种以骨粉为原料由酶法制取明胶的方法 |
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US20220256886A1 (en) | 2022-08-18 |
MX2022005337A (es) | 2022-05-26 |
ZA202205099B (en) | 2023-11-29 |
DE102019130196A1 (de) | 2021-05-12 |
JP2023502010A (ja) | 2023-01-20 |
AU2020378186A1 (en) | 2022-05-26 |
BR112022008115A2 (pt) | 2022-07-19 |
CN114845560A (zh) | 2022-08-02 |
EP4017272A1 (de) | 2022-06-29 |
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