CN111574617A - Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof - Google Patents
Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof Download PDFInfo
- Publication number
- CN111574617A CN111574617A CN202010338683.7A CN202010338683A CN111574617A CN 111574617 A CN111574617 A CN 111574617A CN 202010338683 A CN202010338683 A CN 202010338683A CN 111574617 A CN111574617 A CN 111574617A
- Authority
- CN
- China
- Prior art keywords
- collagen peptide
- preventing
- enzymolysis
- skin
- collagen
- 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.)
- Pending
Links
- 102000008186 Collagen Human genes 0.000 title claims abstract description 68
- 108010035532 Collagen Proteins 0.000 title claims abstract description 68
- 229920001436 collagen Polymers 0.000 title claims abstract description 66
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 59
- 208000001132 Osteoporosis Diseases 0.000 title claims abstract description 25
- 206010039984 Senile osteoporosis Diseases 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 238000001914 filtration Methods 0.000 claims abstract description 24
- 238000010438 heat treatment Methods 0.000 claims abstract description 18
- 102000004190 Enzymes Human genes 0.000 claims abstract description 15
- 108090000790 Enzymes Proteins 0.000 claims abstract description 15
- 239000003513 alkali Substances 0.000 claims abstract description 10
- 238000001694 spray drying Methods 0.000 claims abstract description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 34
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 33
- 238000002791 soaking Methods 0.000 claims description 24
- 239000000243 solution Substances 0.000 claims description 19
- 239000007788 liquid Substances 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000008213 purified water Substances 0.000 claims description 16
- 108090000145 Bacillolysin Proteins 0.000 claims description 15
- 239000000084 colloidal system Substances 0.000 claims description 15
- 241000251468 Actinopterygii Species 0.000 claims description 14
- 229940088598 enzyme Drugs 0.000 claims description 14
- 239000007864 aqueous solution Substances 0.000 claims description 11
- 102000035092 Neutral proteases Human genes 0.000 claims description 10
- 108091005507 Neutral proteases Proteins 0.000 claims description 10
- 238000004140 cleaning Methods 0.000 claims description 10
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 239000012535 impurity Substances 0.000 claims description 8
- 241000283690 Bos taurus Species 0.000 claims description 7
- 238000005520 cutting process Methods 0.000 claims description 7
- 230000008014 freezing Effects 0.000 claims description 7
- 238000007710 freezing Methods 0.000 claims description 7
- 239000004365 Protease Substances 0.000 claims description 6
- 239000010985 leather Substances 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 6
- 238000007790 scraping Methods 0.000 claims description 6
- 230000000415 inactivating effect Effects 0.000 claims description 5
- 241000194108 Bacillus licheniformis Species 0.000 claims description 4
- 108010010803 Gelatin Proteins 0.000 claims description 4
- 102000004157 Hydrolases Human genes 0.000 claims description 4
- 108090000604 Hydrolases Proteins 0.000 claims description 4
- 108090000526 Papain Proteins 0.000 claims description 4
- 108090000631 Trypsin Proteins 0.000 claims description 4
- 102000004142 Trypsin Human genes 0.000 claims description 4
- 230000001877 deodorizing effect Effects 0.000 claims description 4
- 230000002255 enzymatic effect Effects 0.000 claims description 4
- 229920000159 gelatin Polymers 0.000 claims description 4
- 239000008273 gelatin Substances 0.000 claims description 4
- 235000019322 gelatine Nutrition 0.000 claims description 4
- 235000011852 gelatine desserts Nutrition 0.000 claims description 4
- 239000000413 hydrolysate Substances 0.000 claims description 4
- 235000019834 papain Nutrition 0.000 claims description 4
- 229940055729 papain Drugs 0.000 claims description 4
- 239000012588 trypsin Substances 0.000 claims description 4
- 241000282887 Suidae Species 0.000 claims description 3
- 241000131386 Aspergillus sojae Species 0.000 claims description 2
- 101000634587 Bacillus amyloliquefaciens Bacillolysin Proteins 0.000 claims description 2
- 108090000270 Ficain Proteins 0.000 claims description 2
- 108090000284 Pepsin A Proteins 0.000 claims description 2
- 102000057297 Pepsin A Human genes 0.000 claims description 2
- 241000276707 Tilapia Species 0.000 claims description 2
- 239000000872 buffer Substances 0.000 claims description 2
- 230000009849 deactivation Effects 0.000 claims description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 2
- 235000019836 ficin Nutrition 0.000 claims description 2
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 claims description 2
- 230000003301 hydrolyzing effect Effects 0.000 claims description 2
- 229940111202 pepsin Drugs 0.000 claims description 2
- 229960001322 trypsin Drugs 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 10
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 9
- 230000007062 hydrolysis Effects 0.000 abstract description 5
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 5
- 230000009435 amidation Effects 0.000 abstract description 4
- 238000007112 amidation reaction Methods 0.000 abstract description 4
- 229920001184 polypeptide Chemical group 0.000 abstract description 4
- 125000000539 amino acid group Chemical group 0.000 abstract description 3
- 230000004071 biological effect Effects 0.000 abstract description 3
- 230000002779 inactivation Effects 0.000 abstract 1
- 239000000203 mixture Substances 0.000 description 10
- 210000000988 bone and bone Anatomy 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 6
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 6
- 229960002591 hydroxyproline Drugs 0.000 description 6
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 6
- 239000004744 fabric Substances 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 230000037182 bone density Effects 0.000 description 4
- 201000008482 osteoarthritis Diseases 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- 229910052791 calcium Inorganic materials 0.000 description 3
- 239000011575 calcium Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 235000015872 dietary supplement Nutrition 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 208000006820 Arthralgia Diseases 0.000 description 2
- 206010006956 Calcium deficiency Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 206010003246 arthritis Diseases 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000004332 deodorization Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 239000003797 essential amino acid Substances 0.000 description 2
- 235000020510 functional beverage Nutrition 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- 208000006386 Bone Resorption Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 206010060820 Joint injury Diseases 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000002449 bone cell Anatomy 0.000 description 1
- 230000024279 bone resorption Effects 0.000 description 1
- 230000037118 bone strength Effects 0.000 description 1
- CADZRPOVAQTAME-UHFFFAOYSA-L calcium;hydroxy phosphate Chemical compound [Ca+2].OOP([O-])([O-])=O CADZRPOVAQTAME-UHFFFAOYSA-L 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000007691 collagen metabolic process Effects 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 230000008407 joint function Effects 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
Images
Classifications
-
- 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]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physical Education & Sports Medicine (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Wood Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Rheumatology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Gastroenterology & Hepatology (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a collagen peptide for preventing and treating senile osteoporosis and a preparation method thereof, belonging to the technical field of collagen peptide preparation. The preparation method of the collagen peptide comprises the following steps: pretreatment, alkali treatment, pressure heat treatment, filtration for removing hair and fat, enzyme hydrolysis and enzyme inactivation, decoloration, odor removal, filtration, spray drying and the like. The invention uses alkali treatment method to make the isoelectric point of the collagen peptide below 5 and the amidation degree below 5%; by the alkali treatment-enzyme hydrolysis method, macromolecular chains are effectively sheared, effective amino acid groups and polypeptide chains are completely reserved, the content of collagen peptides with the molecular weight of 3000-6000 Da accounts for more than 75%, the obtained polypeptides have biological activity, and the purity of the collagen peptides can reach more than 95%.
Description
Technical Field
The invention relates to a collagen peptide for preventing and treating senile osteoporosis and a preparation method thereof, belonging to the technical field of collagen peptide preparation.
Background
The synthesis of collagen is most vigorous in about 25 years old, the synthesis amount of collagen is gradually reduced with the increase of age, the content of the collagen is only 50% of that of the collagen in the young people at the age of 60 years old, the resistance of body connective tissues is reduced due to the reduction of the collagen, and the collagen metabolism is abnormal, so that chronic orthopedic diseases such as arthralgia, arthritis, rheumatoid arthritis, osteoporosis and the like are caused.
The collagen peptide can relieve Osteoarthritis (OA) symptom, and can repair bone joint injury or slow degeneration without side effect. The collagen peptide can stimulate articular cartilage to produce collagen II, hyaluronic acid, mucopolysaccharide and proteoglycan, inhibit damage of chondrocyte, relieve joint pain of osteoarthritis patients and improve joint function. Collagen peptides also increase the number and differentiation of osteoblasts, reducing osteoclast-mediated bone resorption. In addition, collagen is rich in glycine and amino acids such as proline and hydroxyproline which are generally less in other proteins, and the specific hydroxyproline has a carrying function of conveying calcium in blood to bone cells, while the active collagen is a calcium hydroxy phosphate adhesive, so that calcium can quickly reach bone parts and precipitate to form new bone molecules, thereby improving bone density, enhancing bone strength, preventing osteoporosis, and simultaneously having good prevention and treatment effects on bone diseases such as arthritis and the like.
In recent years, domestic and foreign enterprises add a proper amount of collagen peptide on the basis of the original health food bone strengthening powder in order to improve osteoporosis of middle-aged and elderly people, and the hydroxyproline in the collagen peptide can send calcium in blood to bones for deposition, so that the effects of increasing bones and enhancing bone density are achieved, and meanwhile, the active collagen has good stabilizing effect on co-solubility of water and fat, so that the active collagen and the fat can bring out the best of each other. Foreign researches show that the high-purity collagen can treat early osteoporosis caused by calcium deficiency, and the collagen with the molecular weight of 3000-6000 Da can effectively improve and inhibit the reduction of bone density and maintain the function of bones. However, in the prior art, the prepared collagen peptide has the following problems: the molecular weight is low, mostly below 1000Da, and the molecular weight exists in the form of free amino acid or oligopeptide; the purity is low, and the salt content is high; no biological activity, and is only used as a nutrient supplement of amino acid; the amino acid types are less, and the content of glycine, hydroxyproline/proline and glutamic acid is lower.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a collagen peptide specially for preventing and treating senile osteoarthritis and osteoporosis, which is obtained by treating different skin raw materials (cattle, pigs or fish) by alkali treatment, enzyme hydrolysis, decoloration and deodorization, dialysis and precipitation, filtration and the like.
The technical scheme of the invention is as follows:
a collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) pretreatment: removing surface hair from the wet skin sheet, scraping off a fat layer, cleaning, cutting into pieces, and freezing for later use, wherein the skin sheet is the skin sheet of cattle, pigs or fish; the fish is one or more of tilapia, puffer fish and cod.
(2) Alkali treatment: soaking the pretreated skin slices in NaOH aqueous solution, and cleaning the skin slices with purified water to be neutral after soaking; the mass concentration of the NaOH aqueous solution is 0.3-1%, and the soaking time is 0.5-2 h;
(3) pressure heat treatment: soaking the alkali-treated skin pieces in purified water, placing in an autoclave, and performing heat treatment under pressure;
(4) and (3) filtering to remove hair and fat: filtering the skin sheets subjected to high-temperature and high-pressure treatment to remove hair and impurities, refrigerating at 4-10 ℃ for 12-24 hours, and filtering out fat to obtain a leather colloid;
(5) enzymatic hydrolysis and enzyme deactivation: putting the hide gelatin into a hydrolytic enzyme solution, performing enzymolysis for 0.5-2 hours at 37-50 ℃, heating to over 85 ℃ after the enzymolysis is finished, and inactivating the enzyme to obtain an enzymatic hydrolysate;
(6) decoloring, deodorizing and filtering: after the enzymolysis liquid is cooled to room temperature, adding activated carbon powder, and shaking for decoloring and deodorizing at room temperature; then filtering and removing carbon by using an active carbon filter until the solution is clear and transparent;
(7) spray drying to obtain the collagen peptide.
In the step (2), the mass concentration of the NaOH aqueous solution is 0.5-0.7%; the material-liquid ratio is 1: 5-1: 10g/mL (preferably 1:8 g/mL); the skin piece soaking time is 1-1.5 h.
In the step (3), the feed-liquid ratio of the skin pieces to the purified water is 1: 5-1: 10g/mL, preferably 1:8 g/mL.
In the step (3), the pressure is 0.05-0.1 MPa, preferably 0.08 MPa; the temperature is 110-130 ℃, preferably 115-120 ℃; the heat treatment time is 1-3 h, preferably 1.5-2 h.
In the step (5), the hydrolase is one or more of papain, trypsin, pepsin, ficin and neutral protease. The neutral protease is one or more of bacillus subtilis neutral protease, bacillus licheniformis neutral protease, bacillus amyloliquefaciens neutral protease and aspergillus sojae neutral protease.
In the step (5), the mass concentration of the hydrolase liquid is 0.5-2%, preferably 1%; the enzymolysis temperature is 40-45 ℃; the enzymolysis time is 1-1.5 h, the enzymolysis pH is 2-8, and the enzymolysis pH is preferably 6-7.
In the step (6), the dosage of the activated carbon is 0.5-2%, preferably 1%, of the mass concentration of the activated carbon in the enzymolysis liquid; the decoloring time is 4-12 h, preferably 6-8 h.
The collagen peptide prepared by the preparation method provided by the invention has the beneficial effects that:
the collagen protein with the molecular weight of 3000-6000 Da can treat early osteoporosis caused by calcium deficiency, and the collagen protein with the molecular weight of 3000-6000 Da can effectively improve and inhibit the reduction of bone density and maintain the function of bones;
the method removes impurities by means of decoloration, deodorization, dialysis, precipitation and filtration, improves the purity, and ensures that the purity of the obtained collagen peptide reaches over 95 percent, the salt content is extremely low, and the content of burning residues is 0.5 to 1 percent;
the method has the advantages that the hair is removed and degreased in a high-temperature and high-pressure heat treatment mode, the traditional chemical degreasing mode is different, chemical reagents (such as residues of ethanol, diethyl ether and acetone) are avoided, and the influence on the purity of the final collagen peptide caused by introducing hybrid proteins such as lipase is also avoided.
The collagen peptide completely reserves polypeptide chains of effective amino acid groups, the peptide chains contain dozens of to dozens of amino acids, and sites on the cell surface can specifically identify special amino acid segments, so that the biological activity is exerted, and the cell proliferation and differentiation are promoted;
the collagen peptide contains 8 essential amino acids of 9 essential amino acids, wherein the contents of glycine, hydroxyproline/proline and glutamic acid are very high and account for more than 50 percent of the total amount of the amino acids;
the collagen peptide of the present invention can be incorporated into functional foods and beverages, nutritional skin care products and dietary supplements simply and efficiently at low cost without affecting the taste or odor of the product.
Drawings
FIG. 1 is a process flow diagram of the present invention.
FIG. 2 is a molecular weight distribution diagram of the collagen peptide obtained in example 1.
Detailed Description
The process flow diagram of the present invention is shown in figure 1. The present invention will be described in detail with reference to the accompanying drawings and examples.
Example 1
A collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) extracting surface hair of wet cowhide with shaver, scraping off fat layer with scraper, cleaning, cutting into pieces, and freezing.
(2) Soaking 100g of pretreated cowhide in 1000mL of NaOH aqueous solution (with mass concentration of 0.5%) for 1h, and washing the cowhide with purified water to neutrality after soaking.
(3) Soaking cowhide in purified water (material-liquid ratio of 1:5g/mL), placing in autoclave, and heat treating under pressure at 115 deg.C and 0.05MPa for 3 hr.
(4) Filtering with cloth filter bag to remove hair and impurities, refrigerating at 4 deg.C for 12 hr, and filtering with gauze to remove fat to obtain cowhide gelatin.
(5) Adjusting pH of cow leather colloid to 6.0 + -0.2 with 2M HCl solution, adding 1% (based on dry cow leather colloid) Bacillus subtilis neutral protease, performing enzymolysis at 50 deg.C for 50min, heating to 85 deg.C to inactivate enzyme to obtain enzymatic hydrolysate.
(6) After the enzymolysis liquid is cooled to room temperature, 2 percent of activated carbon powder is added, the mixture is shaken at 80rpm for 6 hours at the room temperature, and after decoloration is finished, the mixture is filtered by an activated carbon filter (a 0.22 mu m filter membrane) to remove carbon until the solution is clear and transparent. Finally spray drying to obtain the collagen peptide.
Example 2
A collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) extracting surface hair from wet pigskin with shaver, scraping off fat layer with scraper, cleaning, cutting into pieces, and freezing.
(2) Soaking 100g of pretreated pigskin in 800mL of NaOH aqueous solution (mass concentration is 0.6%) for 1.2h, and washing the pigskin with purified water to be neutral after soaking.
(3) Soaking pigskin in purified water (feed liquid ratio 1:6g/mL), placing in an autoclave, and performing heat treatment under pressure at 118 deg.C and 0.1MPa for 2.5 h.
(4) Filtering with cloth filter bag to remove hair and impurities, refrigerating at 6 deg.C for 18 hr, and filtering with gauze to remove fat to obtain pigskin colloid.
(5) Adjusting the pH of the pigskin colloid to 7.0 +/-0.2 by using 2M HCl solution, adding 0.5 percent (calculated by the dry product of the pigskin colloid) of trypsin and 0.5 percent of fig enzyme, carrying out enzymolysis for 1h at 45 ℃, heating to 85 ℃ after finishing the enzymolysis, and inactivating the enzyme to obtain an enzymolysis solution.
(6) After the enzymolysis liquid is cooled to room temperature, 1 percent of activated carbon powder is added, the mixture is shaken at 80rpm for 7 hours at the room temperature, and after decoloration is finished, the mixture is filtered by an activated carbon filter (a 0.22 mu m filter membrane) to remove carbon until the solution is clear and transparent. Finally spray drying to obtain the collagen peptide.
Example 3
A collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) cleaning wet fish skin, cutting into pieces, and freezing for later use.
(2) Soaking 100g of pretreated fish skin in 500mL of NaOH aqueous solution (with the mass concentration of 0.5%) for 0.5h, and cleaning the fish skin with purified water to be neutral after soaking.
(3) Soaking fish skin in purified water (feed liquid ratio 1:10g/mL), placing in an autoclave, and performing heat treatment under 120 deg.C and 0.05MPa for 2 hr.
(4) Filtering with a filter cloth bag to remove impurities, refrigerating at 8 deg.C for 24 hr, and filtering with gauze to remove fat to obtain fish skin colloid.
(5) Adjusting the pH of the fish skin colloid to 6.5 +/-0.2 by using 2M HCl solution, adding 0.5 percent of papain and 1 percent of bacillus licheniformis neutral protease (calculated on the dry product of the fish skin colloid), performing enzymolysis for 1.5h at 40 ℃, heating to 85 ℃ after the enzymolysis is finished, and inactivating the enzyme to obtain an enzymolysis solution.
(6) After the enzymolysis liquid is cooled to room temperature, 2 percent of activated carbon powder is added, the mixture is shaken at 80rpm for 10 hours at the room temperature, and after decoloration is finished, the mixture is filtered by an activated carbon filter (a 0.22 mu m filter membrane) to remove carbon until the solution is clear and transparent. Finally spray drying to obtain the collagen peptide.
Example 4
A collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) extracting surface hair of wet cowhide with shaver, scraping off fat layer with scraper, cleaning, cutting into pieces, and freezing.
(2) 100g of pretreated cowhide is soaked in 600mL of NaOH aqueous solution (with the mass concentration of 0.7%) for 1h, and after soaking, the cowhide is washed to be neutral by purified water.
(3) Soaking cowhide in purified water (material-liquid ratio of 1:8g/mL), placing in autoclave, and heat treating under pressure at 115 deg.C and 0.08MPa for 3 hr.
(4) Filtering with cloth filter bag to remove hair and impurities, refrigerating at 10 deg.C for 18 hr, and filtering with gauze to remove fat to obtain cowhide gelatin.
(5) Adjusting pH of cow leather colloid to 6.0 + -0.2 with 2M HCl solution, adding 0.5% (based on dry cow leather colloid) of Bacillus subtilis neutral protease and 0.5% of Bacillus licheniformis neutral protease, performing enzymolysis at 50 deg.C for 1.5 hr, heating to 85 deg.C to inactivate enzyme, and obtaining enzymatic hydrolysate.
(6) After the enzymolysis liquid is cooled to room temperature, 1 percent of activated carbon powder is added, the mixture is shaken at 80rpm for 12 hours at the room temperature, and after decoloration is finished, the mixture is filtered by an activated carbon filter (a 0.22 mu m filter membrane) to remove carbon until the solution is clear and transparent. Finally spray drying to obtain the collagen peptide.
Example 5
A collagen peptide for preventing and treating senile osteoporosis is prepared by the following steps:
(1) extracting surface hair from wet pigskin with shaver, scraping off fat layer with scraper, cleaning, cutting into pieces, and freezing.
(2) Soaking 100g of pretreated pigskin in 1000mL of NaOH aqueous solution (mass concentration is 0.6%) for 2h, and washing the pigskin with purified water to be neutral after soaking.
(3) Soaking pigskin in purified water (feed liquid ratio 1:5g/mL), placing in an autoclave, and performing heat treatment under pressure at 118 deg.C and 0.1MPa for 3 h.
(4) Filtering with cloth filter bag to remove hair and impurities, refrigerating at 4 deg.C for 24 hr, and filtering with gauze to remove fat to obtain pigskin colloid.
(5) Adjusting the pH value of the pigskin colloid to 7.5 +/-0.2 by using 2M HCl solution, adding 0.5 percent (calculated by dry products of the pigskin colloid) of trypsin, 0.5 percent of bacillus subtilis neutral protease and 0.5 percent of papain, carrying out enzymolysis for 50min at 50 ℃, heating to 85 ℃ after the enzymolysis is finished, and inactivating the enzyme to obtain an enzymolysis solution.
(6) After the enzymolysis liquid is cooled to room temperature, 1.5 percent of activated carbon powder is added, the mixture is shaken at 80rpm for 8 hours at room temperature, and after decoloration is finished, the mixture is filtered by an activated carbon filter (0.22 mu m filter membrane) to remove carbon until the solution is clear and transparent. Finally spray drying to obtain the collagen peptide.
Test example 1
The isoelectric point, amidation degree, protein content and hydroxyproline content of the collagen peptides obtained in the examples are shown in Table 1. As is clear from Table 1, the collagen peptide obtained by the method of the present invention using alkali hydrolysis has an isoelectric point of 5 or less and an amidation degree of 5% or less, and the collagen peptide obtained by the method different from acid hydrolysis has an isoelectric point of 6 or more and an amidation degree of 15% or more.
TABLE 1
Test example 2
The molecular weight distribution of the collagen peptide produced under the conditions of example 1 is shown in table 2 and fig. 2. The calculation shows that the content of the collagen peptide with the molecular weight of 3000-6000 Da accounts for more than 75%, which indicates that the method effectively shears a macromolecular chain and most completely retains effective amino acid groups and polypeptide chains through an alkali treatment-enzyme hydrolysis method. The molecular weight distributions of the other examples are shown in Table 3.
TABLE 2
| Retention time | Mn | Mw | MP | Area of | |
|
| 1 | 13.017 | 12491 | 12939 | 9988 | 2262557 | 7.65 |
| 2 | 13.849 | 6725 | 6986 | 6275 | 6674710 | 22.57 |
| 3 | 14.943 | 3770 | 3858 | 3406 | 7035799 | 23.79 |
| 4 | 15.167 | 2516 | 2551 | 3007 | 4509520 | 15.25 |
| 5 | 17.015 | 1355 | 1424 | 1071 | 4320451 | 14.61 |
| 6 | 17.133 | 761 | 786 | 1003 | 3104047 | 10.50 |
| 7 | 18.998 | 322 | 336 | 354 | 1586662 | 5.37 |
| 8 | 20.792 | 135 | 137 | 130 | 77239 | 0.26 |
TABLE 3
| Sample Mw | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| ＞13.5kDa | 0 | 0 | 0 | 0 | 0 |
| 7.5-13.5kDa | 6.65％ | 7.73％ | 7.19％ | 10.20％ | 10.02％ |
| 3.5-7.5kDa | 47.36％ | 47.11％ | 39.46％ | 41.56％ | 45.41％ |
| 1.4-3.5kDa | 29.86％ | 27.45％ | 32.05％ | 31.3％ | 28.2％ |
| 0.5-1.4kDa | 10.5％ | 10.22％ | 13.34％ | 10.81％ | 9.94％ |
| ＜0.5kDa | 5.63％ | 7.50％ | 7.96％ | 6.12％ | 6.42％ |
| Average molecular weight | 4082Da | 4507Da | 3773Da | 4454Da | 4559Da |
Test example 3
Some performance indexes of the collagen peptides prepared in the examples are shown in Table 4. The collagen peptide provided by the invention can be simply and efficiently incorporated into functional foods and beverages, nutritional skin care products and dietary supplements at low cost, and does not affect the taste or smell of the products.
TABLE 4
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. The collagen peptide for preventing and treating senile osteoporosis is characterized in that the preparation method of the collagen peptide comprises the following steps:
(1) pretreatment: removing surface hair from the wet skin sheet, scraping off a fat layer, cleaning, cutting into pieces, and freezing for later use, wherein the skin sheet is a skin sheet of cattle, pigs or fish;
(2) alkali treatment: soaking the pretreated skin slices in NaOH aqueous solution, and cleaning the skin slices with purified water to be neutral after soaking; the mass concentration of the NaOH aqueous solution is 0.3-1%, and the soaking time is 0.5-2 h;
(3) pressure heat treatment: soaking the alkali-treated skin pieces in purified water, placing in an autoclave, and performing heat treatment under pressure;
(4) and (3) filtering to remove hair and fat: filtering the skin sheets subjected to high-temperature and high-pressure treatment to remove hair and impurities, refrigerating at 4-10 ℃ for 12-24 hours, and filtering out fat to obtain a leather colloid;
(5) enzymatic hydrolysis and enzyme deactivation: putting the hide gelatin into a hydrolytic enzyme solution, performing enzymolysis for 0.5-2 hours at 37-50 ℃, heating to over 85 ℃ after the enzymolysis is finished, and inactivating the enzyme to obtain an enzymatic hydrolysate;
(6) decoloring, deodorizing and filtering: after the enzymolysis liquid is cooled to room temperature, adding activated carbon powder, and shaking for decoloring and deodorizing at room temperature; then filtering and removing carbon by using an active carbon filter until the solution is clear and transparent;
(7) spray drying to obtain the collagen peptide.
2. The collagen peptide for preventing and treating senile osteoporosis according to claim 1, wherein in the step (2), the mass concentration of NaOH aqueous solution is 0.5-0.7%; the material-liquid ratio is 1: 5-1: 10 g/mL; the skin piece soaking time is 1-1.5 h.
3. The collagen peptide for preventing and treating senile osteoporosis of claim 1, wherein in the step (3), the feed-liquid ratio of the skin pieces to the purified water is 1: 5-1: 10 g/mL.
4. The collagen peptide for preventing and treating senile osteoporosis according to claim 1, wherein in the step (3), the pressure is 0.05-0.1 MPa; the temperature is 110-130 ℃; the heat treatment time is 1-3 h.
5. The collagen peptide for preventing and treating osteoporosis of the elderly as claimed in claim 1, wherein in step (5), the hydrolase is one or more of papain, trypsin, pepsin, ficin, and neutral protease.
6. The collagen peptide for preventing and treating senile osteoporosis according to claim 5, wherein the neutral protease is one or more of Bacillus subtilis neutral protease, Bacillus licheniformis neutral protease, Bacillus amyloliquefaciens neutral protease and Aspergillus sojae neutral protease.
7. The collagen peptide for preventing and treating senile osteoporosis as claimed in claim 1, wherein in step (5), the mass concentration of the hydrolase solution is 0.5-2%; the enzymolysis temperature is 40-45 ℃; the enzymolysis time is 1-1.5 h, and the enzymolysis pH is 2-8.
8. The collagen peptide for preventing and treating senile osteoporosis as claimed in claim 1, wherein in step (6), the amount of activated carbon is such that the mass concentration of activated carbon in the enzymolysis solution is 0.5% -2%; the decoloring time is 4-12 h.
9. The collagen peptide for preventing and treating osteoporosis of the elderly according to claim 1, wherein in the step (1), the fish is one or more of tilapia, puffer fish and cod.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010338683.7A CN111574617A (en) | 2020-04-26 | 2020-04-26 | Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010338683.7A CN111574617A (en) | 2020-04-26 | 2020-04-26 | Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111574617A true CN111574617A (en) | 2020-08-25 |
Family
ID=72114103
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010338683.7A Pending CN111574617A (en) | 2020-04-26 | 2020-04-26 | Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111574617A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117736307A (en) * | 2024-02-19 | 2024-03-22 | 北京盛美诺生物技术有限公司 | III type small molecule collagen peptide capable of promoting III type and IV type collagen secretion and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101886106A (en) * | 2010-07-02 | 2010-11-17 | 湖北远成药业有限公司 | Method for extracting collagen peptide from fish scales |
| CN101892278A (en) * | 2009-05-18 | 2010-11-24 | 湛江中南岛生化有限公司 | Method for extracting micromolecular collagen oligopeptide from fish |
| WO2014068408A2 (en) * | 2012-10-23 | 2014-05-08 | Caris Life Sciences Switzerland Holdings, S.A.R.L. | Aptamers and uses thereof |
| CN105982337A (en) * | 2016-05-13 | 2016-10-05 | 浙江省海洋开发研究院 | Aquatic collagen peptide food used for adjuvant therapy of osteoporosis and arthritis |
| CN108715612A (en) * | 2018-04-19 | 2018-10-30 | 兰溪市沉默生物科技有限公司 | The preparation method of fish collagen antioxidant peptide |
| CN109504730A (en) * | 2018-12-14 | 2019-03-22 | 金华市艾力生物科技有限公司 | A kind of extracting method of fish-skin active peptides |
-
2020
- 2020-04-26 CN CN202010338683.7A patent/CN111574617A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101892278A (en) * | 2009-05-18 | 2010-11-24 | 湛江中南岛生化有限公司 | Method for extracting micromolecular collagen oligopeptide from fish |
| CN101886106A (en) * | 2010-07-02 | 2010-11-17 | 湖北远成药业有限公司 | Method for extracting collagen peptide from fish scales |
| WO2014068408A2 (en) * | 2012-10-23 | 2014-05-08 | Caris Life Sciences Switzerland Holdings, S.A.R.L. | Aptamers and uses thereof |
| CN105982337A (en) * | 2016-05-13 | 2016-10-05 | 浙江省海洋开发研究院 | Aquatic collagen peptide food used for adjuvant therapy of osteoporosis and arthritis |
| CN108715612A (en) * | 2018-04-19 | 2018-10-30 | 兰溪市沉默生物科技有限公司 | The preparation method of fish collagen antioxidant peptide |
| CN109504730A (en) * | 2018-12-14 | 2019-03-22 | 金华市艾力生物科技有限公司 | A kind of extracting method of fish-skin active peptides |
Non-Patent Citations (1)
| Title |
|---|
| 杨叶辉: "鱼鳞胶原蛋白酶法生产的质量控制体系", 《渔业研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117736307A (en) * | 2024-02-19 | 2024-03-22 | 北京盛美诺生物技术有限公司 | III type small molecule collagen peptide capable of promoting III type and IV type collagen secretion and preparation method thereof |
| CN117736307B (en) * | 2024-02-19 | 2024-06-07 | 北京盛美诺生物技术有限公司 | III type small molecule collagen peptide capable of promoting III type and IV type collagen secretion and preparation method thereof |
| US12391744B1 (en) | 2024-02-19 | 2025-08-19 | Beijing Semnl Biotechnology Co., Ltd. | Micromolecular type III collagen peptide capable of promoting secretion of type III and IV collagens, and preparation method therefor |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106359631B (en) | A kind of pure collagen milk powder and preparation method thereof | |
| CN101061827A (en) | Industry method of producing fish collagen peptide from fish skin and bone by an enzyme method | |
| US11702447B2 (en) | Methods for producing collagen | |
| CN107365824A (en) | A kind of preparation method for hydrolyzing Isin glue collagen Gly-His-Lys | |
| KR101871395B1 (en) | Method for production collagen with high yield | |
| CN112410392A (en) | Extraction method and application of type I collagen | |
| CN110592053A (en) | Collagen hydrolysis complex enzyme, protein oligopeptide and preparation method thereof | |
| JP2023530376A (en) | Method for producing collagen peptide | |
| CN117551186A (en) | Method for preparing collagen peptide by countercurrent immobilized enzyme column enzymolysis | |
| KR100647033B1 (en) | Method for preparing sterilized pure water-soluble collagen peptide | |
| CN111574617A (en) | Collagen peptide for preventing and treating senile osteoporosis and preparation method thereof | |
| CN1582771A (en) | Production of collagen peptide from fish skins | |
| CN115449535A (en) | Bone-derived collagen tripeptide and preparation method thereof | |
| WO2023082523A1 (en) | Method for improving extraction rate of chondroitin sulfate prepared from tilapia skull | |
| CN113754759A (en) | Process for extracting multiple nutritional ingredients from fish scales | |
| CN118360358A (en) | A preparation method of collagen tripeptide, its product and its application in promoting fibroblast proliferation | |
| KR100679712B1 (en) | How to make collagen from starfish | |
| CN119552936A (en) | A fish collagen peptide enzymatic hydrolysis process | |
| JP2004073186A (en) | Method for producing amino acid component by enzymatic degradation of fish egg shell | |
| CN118048425A (en) | Antioxidant collagen peptide and preparation method and application thereof | |
| CN105002246A (en) | Fish polypeptide with uniform molecular weight distribution, and preparation method thereof | |
| Permadi et al. | Characteristics and Applications of Collagen from the Animal By-Product as a Potential Source for Food Ingredients | |
| CN115669792A (en) | Method for preparing debitterized low-molecular-weight mucin by using sturgeon tendon | |
| Jain et al. | Bioprocessing of Chicken Meat and Egg Processing Industries' Waste to Value‐Added Proteins and Peptides | |
| CN113528602A (en) | Salmon skin polypeptide extraction method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200825 |