CN113754759A - Process for extracting multiple nutritional ingredients from fish scales - Google Patents
Process for extracting multiple nutritional ingredients from fish scales Download PDFInfo
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- CN113754759A CN113754759A CN202111087444.XA CN202111087444A CN113754759A CN 113754759 A CN113754759 A CN 113754759A CN 202111087444 A CN202111087444 A CN 202111087444A CN 113754759 A CN113754759 A CN 113754759A
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- acid
- soluble collagen
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- fish scales
- enzyme
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- 241000251468 Actinopterygii Species 0.000 title claims abstract description 131
- 238000000034 method Methods 0.000 title claims abstract description 71
- 230000008569 process Effects 0.000 title claims abstract description 30
- 235000016709 nutrition Nutrition 0.000 title description 7
- 239000004615 ingredient Substances 0.000 title description 6
- 102000008186 Collagen Human genes 0.000 claims abstract description 209
- 108010035532 Collagen Proteins 0.000 claims abstract description 209
- 229920001436 collagen Polymers 0.000 claims abstract description 209
- 102000011782 Keratins Human genes 0.000 claims abstract description 63
- 108010076876 Keratins Proteins 0.000 claims abstract description 63
- 150000007524 organic acids Chemical class 0.000 claims abstract description 57
- 239000002253 acid Substances 0.000 claims abstract description 32
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 31
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000011575 calcium Substances 0.000 claims abstract description 27
- 229910052791 calcium Inorganic materials 0.000 claims abstract description 27
- 102000004190 Enzymes Human genes 0.000 claims abstract description 25
- 108090000790 Enzymes Proteins 0.000 claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 20
- 238000001694 spray drying Methods 0.000 claims abstract description 19
- 230000001954 sterilising effect Effects 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 238000001914 filtration Methods 0.000 claims abstract description 13
- 239000004365 Protease Substances 0.000 claims abstract description 12
- 235000015097 nutrients Nutrition 0.000 claims abstract description 12
- 108091005804 Peptidases Proteins 0.000 claims abstract description 11
- 238000005238 degreasing Methods 0.000 claims abstract description 5
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 87
- 239000002244 precipitate Substances 0.000 claims description 44
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- 108091005658 Basic proteases Proteins 0.000 claims description 34
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 33
- 238000002791 soaking Methods 0.000 claims description 26
- 238000003756 stirring Methods 0.000 claims description 26
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- 238000001728 nano-filtration Methods 0.000 claims description 22
- 229940088598 enzyme Drugs 0.000 claims description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 20
- 230000000694 effects Effects 0.000 claims description 19
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 14
- 230000002255 enzymatic effect Effects 0.000 claims description 13
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000013078 crystal Substances 0.000 claims description 8
- 238000007710 freezing Methods 0.000 claims description 8
- 230000008014 freezing Effects 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 239000000413 hydrolysate Substances 0.000 claims description 8
- 239000004310 lactic acid Substances 0.000 claims description 7
- 235000014655 lactic acid Nutrition 0.000 claims description 7
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 6
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 6
- 239000001630 malic acid Substances 0.000 claims description 6
- 235000011090 malic acid Nutrition 0.000 claims description 6
- 235000015165 citric acid Nutrition 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- 102000029816 Collagenase Human genes 0.000 claims description 4
- 108060005980 Collagenase Proteins 0.000 claims description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 4
- 238000004140 cleaning Methods 0.000 claims description 4
- 229960002424 collagenase Drugs 0.000 claims description 4
- 238000002425 crystallisation Methods 0.000 claims description 4
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- 239000012528 membrane Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 230000007935 neutral effect Effects 0.000 claims description 4
- 238000009928 pasteurization Methods 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 239000007921 spray Substances 0.000 claims description 4
- 235000011054 acetic acid Nutrition 0.000 claims description 3
- 235000011167 hydrochloric acid Nutrition 0.000 claims description 3
- 238000000265 homogenisation Methods 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 18
- 239000012535 impurity Substances 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 107
- 235000018102 proteins Nutrition 0.000 description 17
- 239000000047 product Substances 0.000 description 13
- 239000004744 fabric Substances 0.000 description 12
- 235000001014 amino acid Nutrition 0.000 description 8
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- 102000035195 Peptidases Human genes 0.000 description 7
- 235000019419 proteases Nutrition 0.000 description 7
- 239000000835 fiber Substances 0.000 description 6
- -1 L-form amino acids Chemical class 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 4
- 108010050808 Procollagen Proteins 0.000 description 3
- 210000000988 bone and bone Anatomy 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 3
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 3
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- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 208000001132 Osteoporosis Diseases 0.000 description 2
- 101710093813 Scale keratin Proteins 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
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- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 210000001724 microfibril Anatomy 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000006340 racemization Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241000252230 Ctenopharyngodon idella Species 0.000 description 1
- 241000252233 Cyprinus carpio Species 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical group CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins 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
- 239000004471 Glycine Substances 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-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
- 241001465754 Metazoa Species 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 206010051246 Photodermatosis Diseases 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003276 anti-hypertensive effect Effects 0.000 description 1
- 206010003246 arthritis Diseases 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
- 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
- 230000004071 biological effect Effects 0.000 description 1
- 239000006227 byproduct Substances 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
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Inorganic materials [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 description 1
- 229940069978 calcium supplement Drugs 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000007691 collagen metabolic process Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- YSMODUONRAFBET-UHNVWZDZSA-N erythro-5-hydroxy-L-lysine Chemical group NC[C@H](O)CC[C@H](N)C(O)=O YSMODUONRAFBET-UHNVWZDZSA-N 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 108010007119 flavourzyme Proteins 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000001156 gastric mucosa Anatomy 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
- 235000013402 health food Nutrition 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- GVALZJMUIHGIMD-UHFFFAOYSA-H magnesium phosphate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O GVALZJMUIHGIMD-UHFFFAOYSA-H 0.000 description 1
- 239000004137 magnesium phosphate Substances 0.000 description 1
- 229960002261 magnesium phosphate Drugs 0.000 description 1
- 229910000157 magnesium phosphate Inorganic materials 0.000 description 1
- 235000010994 magnesium phosphates Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000008845 photoaging Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000009993 protective function Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
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
- 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
- A23L33/16—Inorganic salts, minerals or trace elements
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/461—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from fish
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Toxicology (AREA)
- Wood Science & Technology (AREA)
- Inorganic Chemistry (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
A process for extracting various nutrient components from fish scales comprises the following steps: (1) removing impurity protein from fish scales; (2) degreasing fish scales; (3) decoloring fish scales; (4) removing ash from fish scales to obtain an organic acid calcium solution and decalcified fish scales; (5) crystallizing organic acid calcium; (6) carrying out acid method and enzyme method gel extraction on the decalcified fish scales to obtain an acid soluble collagen solution, an enzyme soluble collagen solution and residues; (7) performing enzymolysis on collagen to obtain acid-soluble collagen enzymolysis liquid and enzyme-soluble collagen enzymolysis liquid; (8) nano-filtering to obtain acid-soluble collagen enzymolysis concentrated solution and enzyme-soluble collagen protease concentrated solution; (9) sterilizing the acid soluble collagen enzymolysis concentrated solution and the enzyme soluble collagen protease concentrated solution, and spray drying to obtain acid soluble collagen peptide powder and enzyme soluble collagen peptide powder; (10) separating the residue by an alkaline method to obtain the keratin. The invention can obtain organic acid calcium, collagen peptide and keratin, and realize full utilization of fish scales.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a process for extracting various nutritional ingredients from fish scales.
Background
The fish scales are byproducts generated in the processing process of scaled fishes, more than 100 million tons of fish scales can be generated in China every year, most of the fish scales are discarded except part of the fish scales are used as feed raw materials, resources are wasted, and environmental pollution is caused. The fish scale collagen has the functions of resisting oxidation, improving the immunity of the organism and the like; keratin can be used as a good raw material of medical biomaterials; the hydroxyapatite extracted from fish scales has good biocompatibility, is a good calcium supplement raw material, and can also be used as a scaffold material of bones or teeth.
Fish scales are rich in protein and various minerals, and mainly consist of protein and hydroxyapatite. The protein accounts for 50-70% of the total weight of the fish scales, is mainly collagen and keratin, and also contains a small amount of globulin, mucin and the like. Collagen is mainly distributed in the inner layer of fish scales, and collagen with an integral structure is fibrin bone scales and generally divided into two layers: the upper layer is a bone layer, the main component of the upper layer is hydroxyapatite, and a small amount of inorganic salts such as calcium carbonate, magnesium phosphate, sodium phosphate and the like are also contained, and collagen is scattered; the lower layer is a fibrous layer, collagen fibers are closely and parallelly arranged in the same thin layer, and the lower layer and the collagen fibers in the adjacent thin layers form a splint structure with different included angles.
The fish scale collagen has good biological characteristics and is closely related to cell proliferation, differentiation, movement, immunity, joint lubrication, wound healing and the like. The collagen peptide has good processing characteristics and physiological effects, meets the requirements of people on low-fat and high-protein foods, and can be used for developing various health-care products. Especially, it has effects of protecting gastric mucosa, resisting ulcer, promoting skin collagen metabolism, preventing arthritis and osteoporosis, and can be used for developing skin caring beverage and health food for preventing osteoporosis. The collagen peptide can be added into sports beverage due to its rapid absorption and high absorption rate.
The collagen peptide has good biocompatibility, good anti-aging activity, inhibitory activity, chronic antihypertensive effect, anti-skin photoaging effect and other biological activities, so that the collagen peptide has good prospects in the application of the biomedical field.
Keratin is a kind of fibrous scleroprotein with connective and protective functions, belongs to structural protein of ectodermal cells, has stable property, is insoluble in water, salt solution, dilute acid or dilute alkali, has strong stretch-resisting performance, is rich in a large amount of cysteine residues, and plays a protective role in animal bodies. The natural keratin has the characteristics of good biocompatibility, bioactivity, biodegradability, excellent material mechanical property, natural abundance and the like, and is widely applied to the fields of textiles, biological materials, packaging materials, medicine, cosmetics and the like.
The prior art is not sufficient in utilization of fish scales, and most of the fish scales only utilize one or two of collagen, keratin and calcium. Chinese patent application publication No. CN106834397A discloses a method for preparing fish scale keratin, according to which collagen and keratin can be obtained, but the application does not relate to how to separate collagen, and keratin is obtained by enzymatic hydrolysis, although the keratin structure is completely preserved, but the yield is too low.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a process for extracting various nutritional ingredients from fish scales, which can extract collagen, keratin and calcium contained in the fish scales, so that the full utilization of the fish scales is realized. The technical scheme is as follows:
a process for extracting various nutrient components from fish scales is characterized by comprising the following steps:
(1) fish scale dehybridization protein
After cleaning fresh fish scales, soaking the fresh fish scales in a sodium chloride solution with the weight percentage concentration of 4-10% at 4-12 ℃ for 8-12 hours, taking out the fish scales, and rinsing the fish scales with clean water;
(2) degreasing of fish scales
Soaking the fish scales after the foreign protein removal in the step (1) in a sodium hydroxide solution with the weight percentage concentration of 0.4-0.8% at 4-12 ℃ for 4-6 hours, then taking out the fish scales, and rinsing the fish scales with clean water;
(3) decolouring fish scales
Soaking the degreased fish scales in the step (2) in a hydrogen peroxide solution with the weight percentage concentration of 1-2% for 4-8 hours at 4-12 ℃, taking out, and rinsing with clean water;
(4) fish scale deashing
Soaking the fish scales decolorized in the step (3) in an organic acid solution at 60-75 ℃ for 4-8 hours under the condition of stirring, standing after stirring, and separating a supernatant from a precipitate, wherein the supernatant is an organic acid calcium solution, and the precipitate is decalcified fish scales;
(5) organic acid calcium crystal
Freezing and crystallizing the supernatant obtained in the step (4) to obtain a crystal, namely the organic acid calcium;
(6) acid method and enzyme method for extracting gel
Adding the decalcified fish scales obtained in the step (4) into an organic acid solution, soaking at 4-8 ℃, and separating a dissolved solution from a precipitate after soaking is finished to obtain an acid-soluble collagen solution;
adding water into the precipitate to prepare a feed liquid, adding alkaline protease for enzymolysis to obtain an enzyme-soluble collagen solution, and separating the enzyme-soluble collagen solution to obtain a residue;
(7) collagen enzymolysis
Respectively carrying out enzymolysis on the acid-soluble collagen solution and the enzyme-soluble collagen solution obtained in the step (6) to obtain an acid-soluble collagen enzymolysis solution and an enzyme-soluble collagen enzymolysis solution;
(8) nanofiltration
Carrying out nanofiltration on the acid-soluble collagen enzymatic hydrolysate and the enzyme-soluble collagen enzymatic hydrolysate obtained in the step (7) respectively to obtain an acid-soluble collagen enzymatic concentrated solution and an enzyme-soluble collagen protease concentrated solution;
(9) sterilization
Respectively sterilizing the acid soluble collagen enzymolysis concentrated solution and the enzyme soluble collagenase concentrated solution obtained in the step (8);
(10) spray drying
Respectively carrying out spray drying on the acid-soluble collagen enzymolysis concentrated solution and the enzyme-soluble collagen protease concentrated solution obtained in the step (10) by using a spray dryer to respectively obtain acid-soluble collagen peptide powder and enzyme-soluble collagen peptide powder;
(11) alkaline process for the separation of keratin
And (3) mixing the residue obtained in the step (6) with a sodium hydroxide solution, reacting at 55-65 ℃ until the residue is completely dissolved, adding concentrated hydrochloric acid to adjust the pH value to the isoelectric point of keratin, standing, and separating out a precipitate, wherein the precipitate is the keratin.
In a preferable scheme, dissolving the precipitate separated in the step (11) by using a sodium hydroxide solution with the weight percentage concentration of 1-3%, and adjusting to be neutral by using hydrochloric acid to obtain a keratin solution; then carrying out nanofiltration on the obtained keratin solution to obtain a keratin concentrated solution; and sterilizing the keratin concentrated solution, and spray drying to obtain dry powdery keratin.
In another preferred embodiment, the precipitate separated in step (11) is freeze-dried to obtain dried keratin.
The step (1) plays a role in removing pigments attached to the fish scales and impure proteins of non-collagen components. Preferably, the rinsing in step (1) is carried out three times by using clear water.
And (3) removing fish scale fat in the step (2). Preferably, the rinsing in step (2) is carried out three times by using clear water.
And (4) decoloring the fish scales. Preferably, the rinsing in step (3) is carried out three times by using clear water.
And (4) removing calcium from the fish scales. Preferably, in the step (4), the organic acid solution has a concentration of 3-10% by weight, and the organic acid is one or a combination of citric acid, malic acid and lactic acid. Preferably, in the step (4), the weight ratio of the decolored fish scales to the organic acid solution is 1: 8-1: 20. Preferably, in the step (4), the stirring is performed at a rotation speed of 30-50 rpm.
Preferably, in the step (5), the freezing crystallization temperature is-8 to-12 ℃, and the crystallization time is 6 to 12 hours.
Preferably, in the step (6), the organic acid solution has a concentration of 5-10% by weight, and the organic acid is one or a combination of acetic acid, citric acid, malic acid and lactic acid. In the preferable step (6), the weight ratio of the deashed fish scales to the organic acid solution is 1: 8-1: 20, and the soaking time is 18-24 hours. The acid-soluble collagen is extracted by breaking ionic bonds between collagen and other substances and amide bonds inside collagen by adding acid. The collagen extracted by the acid method can better keep the three-dimensional structure. The acid method for extracting the collagen can only obtain a part of the collagen when the acid amount is too low, and the collagen is easily damaged when the acid amount is too large, so that the extraction rate is reduced. In the step (6), part of the collagen is extracted with a lower acid consumption (the collagen can be prevented from being damaged) and the rest of the collagen is further extracted by an enzyme method, so that the yield is improved. In addition, the acid extraction can also be performed by hydrochloric acid.
In the preferable step (6), the weight ratio of the precipitate to water is 1: 8-1: 20; the weight of the added alkaline protease is 1-3% of that of the precipitate, and the enzymolysis is carried out for 12-24 hours at 25-35 ℃. The residue obtained in the step (6) contains fish scale keratin. Preferably, the activity unit of the alkaline protease used in the step (6) is 30-80 ten thousand units. Collagen is a protein having a triple-helical structure and has a relative molecular weight of about 30000Da, in which glycine accounts for almost one third of the total amino acid residues and contains hydroxyproline and hydroxylysine residues, which are more rare in other proteins. The collagen molecule unit is called procollagen, and each procollagen molecule consists of three alpha-peptide chains, which form a triple helix with the same axis as the center under the interaction of amino acids. The procollagen molecules are arranged in parallel to form bundles, and through covalent cross-linking, stable collagen microfibrils can be formed, and further the stable collagen microfibrils are aggregated to form bundles, and collagen fibers are formed, and the collagen molecules become insoluble fibers through intramolecular or intermolecular cross-linking. The natural structure of collagen assumes the form of non-helical region-non-helical region, which is responsible for the insolubility of collagen in water. The non-spiral areas at the two ends of the collagen are cut off by adopting protease, so that the enzyme-soluble collagen can be obtained, and the commonly used enzymes comprise pepsin, papain, alkaline protease, compound flavourzyme, trypsin, neutral protease and the like.
In the preferable step (7), adding an acid-soluble collagen solution into a first enzymolysis tank, adding water to adjust the weight percentage concentration of the acid-soluble collagen to be 8-15%, using a ball mill to carry out homogenization, using sodium hydroxide to adjust the pH value to be 7.5-9, heating to 45-60 ℃, adding alkaline protease and neutral protease, wherein the weight of the alkaline protease is 1-3% of that of the acid-soluble collagen, and the weight of the neutral protease is 1-3% of that of the acid-soluble collagen; and then carrying out enzymolysis for 4-8 hours under the condition of stirring (preferably, the stirring rotating speed is 30-50 rpm), and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely the acid-soluble collagen enzymolysis liquid. The enzymolysis product of the acid-soluble collagen solution can be filtered by a cloth bag filter to obtain an enzymolysis supernatant. Preferably, the aperture of a filter bag of the used cloth bag filter is 10-50 microns.
In the preferable step (7), adding the enzyme-soluble collagen into a second enzymolysis tank, adding water to adjust the weight percentage concentration of the enzyme-soluble collagen to be 8-15%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to be 7.5-9, heating to 45-60 ℃, adding alkaline protease and neutral protease, wherein the weight of the alkaline protease is 1-3% of that of the enzyme-soluble collagen, and the weight of the neutral protease is 1-3% of that of the enzyme-soluble collagen; and then carrying out enzymolysis for 4-8 hours under the condition of stirring (preferably, the stirring speed is 30-50 rpm), and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely the enzyme-soluble collagen enzymolysis liquid. The enzymolysis product of the enzyme soluble collagen can be filtered by a cloth bag filter to obtain the enzymolysis supernatant. Preferably, the aperture of a filter bag of the used cloth bag filter is 10-50 microns.
And (4) performing enzymolysis on the collagen in the step (7), wherein the product after the enzymolysis of the collagen is collagen peptide, the triple-helix structure of the collagen is thoroughly loosened and further denatured, and the generated dispersed peptide segment is degraded, and the relative molecular mass of the peptide segment is from hundreds to thousands of daltons.
Preferably, the activity unit of the alkaline protease used in the step (7) is 30-80 ten thousand units, and the activity unit of the neutral protease is 10-30 ten thousand units.
Preferably, the aperture of the nanofiltration membrane used in the nanofiltration in the step (8) is 150-300 nanometers.
In the preferable step (9), the sterilization mode is pasteurization, and the temperature is kept at 80-85 ℃ for 20-30 minutes.
In the preferable step (10), the inlet air temperature of the spray drying is 150-185 ℃, and the outlet air temperature is 60-85 ℃.
In the step (11), the keratin is precipitated at the isoelectric point, and the separation and purification of the keratin can be realized. Preferably, in the step (11), the weight percentage concentration of the sodium hydroxide solution is 4-8%, and the weight of the sodium hydroxide solution is 10-20 times of that of the residue. Preferably, in the step (11), the standing time is 30-60 minutes, and after standing, a centrifugal machine is adopted to separate out precipitates, wherein the rotating speed of the centrifugal machine is 3000-5000 rpm, and the centrifugal time is 10-30 minutes.
Compared with the prior art, the invention has the following beneficial effects:
1. extraction of collagen
At present, the extraction methods of collagen mainly comprise a hot water method, an acid method, an alkaline method and an enzyme method.
The hot water extraction method has low salt content, but has long hydrolysis time, needs to be operated under pressure, and has uneven relative molecular mass distribution of the product and difficult control.
In the alkaline extraction, alkaline solution can be saponified with fat combined with insoluble collagen in connective tissue, non-helical non-terminal peptide is cut off, collagen fiber is disintegrated and dissolved out, and common alkaline treatment agent includes calcium hydroxide, sodium hydroxide, etc. In the alkaline extraction process, due to collagen denaturation and partial hydrolysis of peptide bonds, the supercoiled structure of the collagen is damaged, the prepared collagen has lower molecular weight, and the hydrolysis of the peptide bonds causes asparagine and glutamine to be converted into aspartic acid and glutamic acid, so that the isoelectric point of the collagen is reduced. If the alkaline hydrolysis is severe, racemization of amino acids occurs, forming a racemic mixture of D-form and L-form amino acids, and when the content of D-form amino acids is higher than that of L-form amino acids, absorption of L-form amino acids is inhibited, and most of D-form amino acids have Kazaki-inducing or carcinogenic toxicity.
The common acid treating agent for extracting collagen by acid method comprises hydrochloric acid, acetic acid, citric acid, formic acid, lactic acid, etc., and mainly dissolves collagen without crosslinking or containing amide crosslinking bond, and the obtained product is acid soluble collagen. The kind, concentration, extraction temperature and extraction time of the acid have great influence on the extraction effect of the collagen. The extraction temperature has obvious influence on the molecular weight of the collagen finished product, and the molecular mass distribution of the collagen finished product is wider and is dispersively distributed along with the rise of the temperature. The extraction time only affects the yield of the collagen and has no influence on the molecular weight. The collagen obtained by the method has the performances of higher water retention, emulsion stability, foam stability and the like.
The enzyme method is a method for specifically destroying chemical bonds in collagen fiber molecules under the action of enzyme to obtain collagen with an H-helix structure and a complete preservation. The enzymolysis reaction condition is mild, the reaction rate is high, the time is short, racemization is not generated, the characteristics of amino acid are not influenced, the purity is high, and the safety is good.
The method adopts the acid method and the enzyme method to extract the collagen in the fish scales in two steps, on one hand, the collagen contained in the fish scales can be more fully extracted by utilizing different extraction principles, so that the yield of the collagen is improved, and on the other hand, the collagen obtained by the acid method and the enzyme method has better performance.
2. Extraction of keratin
The methods for extracting keratin at present mainly comprise an enzymatic method, a reduction method and an alkaline method.
The enzymatic method adopts alkaline protease method to extract keratin, although the keratin structure is completely preserved, the yield is too low.
The keratin is extracted by a reduction method, although the yield is slightly higher than that of an alkaline method, the structure is completely stored, but the purification is difficult, and the lauryl calcium sulfate in the reduction method has strong foamability and is difficult to remove in dialysis.
The invention adopts an alkaline method to extract keratin from the residue left after extracting collagen, the extraction time is short, and the product structure is completely preserved.
3. Extraction of calcium
The organic acid calcium with good water solubility is obtained by deliming and crystallizing by an organic acid method. The organic calcium is a composite organic calcium salt formed by chelating calcium ions and organic acid ions, has special spatial structure and bioactivity, good solubility and high bioavailability, is beneficial to absorption of other mineral substances, has good flavor, is a good organic calcium nutrition enhancer, and has very wide application prospect in the aspect of food development.
In short, the invention can obtain organic acid calcium, collagen peptide and keratin by organic acid deliming, acid method and enzyme method extraction of collagen and alkali method extraction of keratin, and further divide the collagen into acid soluble collagen and enzyme soluble collagen, and respectively obtain acid soluble collagen peptide powder and enzyme soluble collagen peptide powder, thereby realizing the full utilization of fish scales.
Detailed Description
Example 1
In this embodiment, the process for extracting various nutritional ingredients from fish scales includes the following steps:
(1) fish scale dehybridization protein
After cleaning fresh fish scales (in the embodiment, fresh carp fish scales), soaking the fresh fish scales in a sodium chloride solution with the weight percentage concentration of 6% at 4 ℃ for 8 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with the clear water for three times);
(2) degreasing of fish scales
Soaking the fish scales subjected to the impurity protein removal in the step (1) in a sodium hydroxide solution with the weight percentage concentration of 0.5% at 4 ℃ for 6 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with the clear water for three times);
(3) decolouring fish scales
Soaking the fish scales degreased in the step (2) in a hydrogen peroxide solution with the weight percentage concentration of 2% at 4 ℃ for 4 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with the clear water for three times);
(4) fish scale deashing
Soaking the fish scales decolorized in the step (3) in an organic acid solution at 65 ℃ for 8 hours under the condition of stirring (stirring at the rotating speed of 40 revolutions per minute), standing after stirring is finished, and separating a supernatant from a precipitate, wherein the supernatant is an organic acid calcium solution, and the precipitate is decalcified fish scales;
in the step (4), the organic acid solution has a weight percentage concentration of 4%, and the organic acid is citric acid and malic acid (the weight ratio of citric acid to malic acid is 1: 2); the weight ratio of the decolored fish scales to the organic acid solution is 1: 10;
(5) organic acid calcium crystal
Freezing and crystallizing the supernatant obtained in the step (4) (the freezing and crystallizing temperature is-12 ℃, and the crystallizing time is 12 hours), wherein the obtained crystal is organic acid calcium;
(6) acid method and enzyme method for extracting gel
Adding the decalcified fish scales obtained in the step (4) into an organic acid solution, soaking for 24 hours at 4 ℃, and separating a dissolved solution from a precipitate after soaking to obtain an acid-soluble collagen solution; adding water into the precipitate to prepare a feed liquid, adding alkaline protease for enzymolysis to obtain an enzyme-soluble collagen solution, and separating the enzyme-soluble collagen solution to obtain a residue;
in the step (6), the weight percentage concentration of the organic acid solution is 8%, and the organic acid contained in the organic acid solution is citric acid; the weight ratio of the deashed fish scales to the organic acid solution is 1: 10;
in the step (6), the weight ratio of the precipitate to the water is 1: 12; adding alkaline protease 1 wt% of the precipitate, and performing enzymolysis at 35 deg.C for 12 hr; the activity unit of the alkaline protease is 80 ten thousand units;
(7) collagen enzymolysis
Respectively carrying out enzymolysis on the acid-soluble collagen solution and the enzyme-soluble collagen solution obtained in the step (6) to obtain an acid-soluble collagen enzymolysis solution and an enzyme-soluble collagen enzymolysis solution;
in the step (7), adding an acid-soluble collagen solution into a first enzymolysis tank, adding water to adjust the weight percentage concentration of the acid-soluble collagen to be 8%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to be 7.5, heating to 52 ℃, adding alkaline protease and neutral protease (the activity unit of the alkaline protease is 50 ten thousand units, the activity unit of the neutral protease is 20 ten thousand units), wherein the weight of the alkaline protease is 3% of that of the acid-soluble collagen, and the weight of the neutral protease is 1% of that of the acid-soluble collagen; then carrying out enzymolysis for 8 hours under the condition of stirring (the stirring speed is 50 rpm), and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely the acid-soluble collagen enzymolysis liquid. And filtering the enzymolysis product of the acid-soluble collagen solution by using a cloth bag filter to obtain an enzymolysis supernatant. The aperture of the filter bag of the cloth bag filter is 30 microns;
in the step (7), adding the enzyme-soluble collagen into a second enzymolysis tank, adding water to adjust the weight percentage concentration of the enzyme-soluble collagen to be 8%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to be 7.5, heating to 52 ℃, adding alkaline protease and neutral protease (the activity unit of the alkaline protease is 50 ten thousand units, the activity unit of the neutral protease is 20 ten thousand units), wherein the weight of the alkaline protease is 3% of that of the enzyme-soluble collagen, and the weight of the neutral protease is 1% of that of the enzyme-soluble collagen; then carrying out enzymolysis for 8 hours under the condition of stirring (the stirring speed is 50 rpm), and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely the enzymatic soluble collagen enzymolysis liquid. And filtering the enzymolysis product of the enzyme-soluble collagen by using a cloth bag filter to obtain an enzymolysis supernatant. The aperture of the filter bag of the cloth bag filter is 30 microns;
(8) nanofiltration
Carrying out nanofiltration (the aperture of a nanofiltration membrane used for nanofiltration is 200 nanometers) on the acid-soluble collagen enzymatic hydrolysate and the enzyme-soluble collagen enzymatic hydrolysate obtained in the step (7) respectively to obtain an acid-soluble collagen enzymatic concentrated solution and an enzyme-soluble collagen protease concentrated solution;
(9) sterilization
Respectively sterilizing the acid soluble collagen enzymolysis concentrated solution and the enzyme soluble collagenase concentrated solution obtained in the step (8) (the sterilization mode is pasteurization, and the solution is kept at 85 ℃ for 30 minutes);
(10) spray drying
Respectively carrying out spray drying on the acid-soluble collagen enzymolysis concentrated solution and the enzyme-soluble collagen protease concentrated solution obtained in the step (10) by using a spray dryer (the air inlet temperature of the spray drying is 172 ℃, the air outlet temperature is 75 ℃), and respectively obtaining acid-soluble collagen peptide powder and enzyme-soluble collagen peptide powder;
(11) alkaline process for the separation of keratin
Mixing the residue obtained in the step (6) with a sodium hydroxide solution, reacting at 65 ℃ until the residue is completely dissolved, then adding concentrated hydrochloric acid to adjust the pH value to the keratin isoelectric point (adjusting the pH value to 4.5), standing, and separating out a precipitate, wherein the precipitate is keratin;
in the step (11), the weight percentage concentration of the sodium hydroxide solution is 5%, and the weight of the sodium hydroxide solution is 12 times of that of the residue; standing for 30 minutes, and separating out precipitates by adopting a centrifugal machine after standing, wherein the rotating speed of the centrifugal machine is 4000rpm, and the centrifugal time is 20 minutes.
Dissolving the precipitate separated in the step (11) by using a sodium hydroxide solution with the weight percentage concentration of 2%, and then adjusting the solution to be neutral by using hydrochloric acid to obtain a keratin solution; then carrying out nanofiltration on the obtained keratin solution to obtain a keratin concentrated solution; and sterilizing the keratin concentrated solution, and spray drying to obtain dry powdery keratin. Nanofiltration, sterilization, and spray drying of the keratin solution were performed according to the above-described steps (8) to (10).
In this example, the yield of the organic acid calcium was 8.7%, the yield of the acid-soluble collagen was 6.4%, the yield of the enzyme-soluble collagen was 10.2%, and the yield of the keratin was 3.7%.
Through detection, the content of protein peptide with the molecular weight of below 1000Da in the acid-soluble collagen peptide powder is 94.2 percent; the content of protein peptide with molecular weight below 1000Da in the enzyme-soluble collagen peptide powder is 95.3%.
In other embodiments, the precipitate separated in step (11) may also be freeze-dried to obtain dried keratin.
Example 2
In this embodiment, the process for extracting various nutritional ingredients from fish scales includes the following steps:
(1) fish scale dehybridization protein
After cleaning fresh fish scales (in this embodiment, fresh grass carp scales), soaking the fresh fish scales in a sodium chloride solution with a weight percentage concentration of 5% at 8 ℃ for 12 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with clear water for three times);
(2) degreasing of fish scales
Soaking the fish scales subjected to the impurity protein removal in the step (1) in a sodium hydroxide solution with the weight percentage concentration of 0.6% at 8 ℃ for 4 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with the clear water for three times);
(3) decolouring fish scales
Soaking the fish scales degreased in the step (2) in a hydrogen peroxide solution with the weight percentage concentration of 1.5% at 8 ℃ for 5 hours, taking out the fish scales, and rinsing the fish scales with clear water (rinsing the fish scales with the clear water for three times);
(4) fish scale deashing
Soaking the fish scales decolorized in the step (3) in an organic acid solution at 65 ℃ for 8 hours under the condition of stirring (stirring at the rotating speed of 30 revolutions per minute), standing after stirring is finished, and separating a supernatant from a precipitate, wherein the supernatant is an organic acid calcium solution, and the precipitate is decalcified fish scales;
in the step (4), the organic acid solution has a weight percentage concentration of 5%, and the organic acid is citric acid and lactic acid (the weight ratio of the citric acid to the lactic acid is 2: 1); the weight ratio of the decolored fish scales to the organic acid solution is 1: 12;
(5) organic acid calcium crystal
Freezing and crystallizing the supernatant obtained in the step (4) (the freezing and crystallizing temperature is-12 ℃, and the crystallizing time is 12 hours), wherein the obtained crystal is organic acid calcium;
(6) acid method and enzyme method for extracting gel
Adding the decalcified fish scales obtained in the step (4) into an organic acid solution, soaking for 24 hours at 4 ℃, and separating a dissolved solution from a precipitate after soaking to obtain an acid-soluble collagen solution; adding water into the precipitate to prepare a feed liquid, adding alkaline protease for enzymolysis to obtain an enzyme-soluble collagen solution, and separating the enzyme-soluble collagen solution to obtain a residue;
in the step (6), the weight percentage concentration of the organic acid solution is 8%, and the organic acid contained in the organic acid solution is citric acid; the weight ratio of the deashed fish scales to the organic acid solution is 1: 10;
in the step (6), the weight ratio of the precipitate to the water is 1: 10; adding alkaline protease 1 wt% of the precipitate, and performing enzymolysis at 25 deg.C for 12 hr; the activity unit of the alkaline protease is 80 ten thousand units;
(7) collagen enzymolysis
Respectively carrying out enzymolysis on the acid-soluble collagen solution and the enzyme-soluble collagen solution obtained in the step (6) to obtain an acid-soluble collagen enzymolysis solution and an enzyme-soluble collagen enzymolysis solution;
in the step (7), adding an acid-soluble collagen solution into a first enzymolysis tank, adding water to adjust the weight percentage concentration of the acid-soluble collagen to be 10%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to be 7.5, heating to 52 ℃, adding alkaline protease and neutral protease (the activity unit of the alkaline protease is 50 ten thousand units, the activity unit of the neutral protease is 20 ten thousand units), wherein the weight of the alkaline protease is 3% of that of the acid-soluble collagen, and the weight of the neutral protease is 1.5% of that of the acid-soluble collagen; then enzymolysis is carried out for 6 hours under the condition of stirring (the stirring speed is 50 rpm), and after the enzymolysis is finished, the enzymolysis supernatant fluid is obtained by filtering, namely the acid-soluble collagen enzymolysis liquid. And filtering the enzymolysis product of the acid-soluble collagen solution by using a cloth bag filter to obtain an enzymolysis supernatant. The aperture of the filter bag of the cloth bag filter is 30 microns;
in the step (7), adding the enzyme-soluble collagen into a second enzymolysis tank, adding water to adjust the weight percentage concentration of the enzyme-soluble collagen to be 10%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to be 7.5, heating to 52 ℃, adding alkaline protease and neutral protease (the activity unit of the alkaline protease is 50 ten thousand units, the activity unit of the neutral protease is 20 ten thousand units), wherein the weight of the alkaline protease is 3% of that of the enzyme-soluble collagen, and the weight of the neutral protease is 1.5% of that of the enzyme-soluble collagen; then enzymolysis is carried out for 6 hours under the condition of stirring (the stirring speed is 50 rpm), and after the enzymolysis is finished, the enzymolysis supernatant fluid is obtained by filtering, namely the enzyme-soluble collagen enzymolysis liquid. And filtering the enzymolysis product of the enzyme-soluble collagen by using a cloth bag filter to obtain an enzymolysis supernatant. The aperture of the filter bag of the cloth bag filter is 30 microns;
(8) nanofiltration
Carrying out nanofiltration (the aperture of a nanofiltration membrane used for nanofiltration is 200 nanometers) on the acid-soluble collagen enzymatic hydrolysate and the enzyme-soluble collagen enzymatic hydrolysate obtained in the step (7) respectively to obtain an acid-soluble collagen enzymatic concentrated solution and an enzyme-soluble collagen protease concentrated solution;
(9) sterilization
Respectively sterilizing the acid soluble collagen enzymolysis concentrated solution and the enzyme soluble collagenase concentrated solution obtained in the step (8) (the sterilization mode is pasteurization, and the solution is kept at 85 ℃ for 30 minutes);
(10) spray drying
Respectively carrying out spray drying on the acid-soluble collagen enzymolysis concentrated solution and the enzyme-soluble collagen protease concentrated solution obtained in the step (10) by using a spray dryer (the air inlet temperature of the spray drying is 178 ℃, the air outlet temperature is 76 ℃), and respectively obtaining acid-soluble collagen peptide powder and enzyme-soluble collagen peptide powder;
(11) alkaline process for the separation of keratin
Mixing the residue obtained in the step (6) with a sodium hydroxide solution, reacting at 60 ℃ until the residue is completely dissolved, then adding concentrated hydrochloric acid to adjust the pH value to the keratin isoelectric point (adjusting the pH value to 4.6), standing, and separating out a precipitate, wherein the precipitate is keratin;
in the step (11), the weight percentage concentration of the sodium hydroxide solution is 5%, and the weight of the sodium hydroxide solution is 10 times of that of the residue; standing for 30 minutes, and separating out precipitates by adopting a centrifugal machine after standing, wherein the rotating speed of the centrifugal machine is 4000rpm, and the centrifugal time is 20 minutes.
Dissolving the precipitate separated in the step (11) by using a sodium hydroxide solution with the weight percentage concentration of 3%, and then adjusting the solution to be neutral by using hydrochloric acid to obtain a keratin solution; then carrying out nanofiltration on the obtained keratin solution to obtain a keratin concentrated solution; and sterilizing the keratin concentrated solution, and spray drying to obtain dry powdery keratin. Nanofiltration, sterilization, and spray drying of the keratin solution were performed according to the above-described steps (8) to (10).
In this example, the yield of the organic acid calcium was 7.9%, the yield of the acid-soluble collagen was 6.9%, the yield of the enzyme-soluble collagen was 12.2%, and the yield of the keratin was 4.2%.
Through detection, the content of protein peptide with the molecular weight of less than 1000Da in the acid-soluble collagen peptide powder is 93.1 percent; the content of protein peptide with molecular weight below 1000Da in the enzyme-soluble collagen peptide powder is 95.6%.
In other embodiments, the precipitate separated in step (11) may also be freeze-dried to obtain dried keratin.
The organic acid calcium yield = organic acid calcium weight/fish scale weight × 100%. Acid soluble collagen yield = acid soluble collagen weight/fish scale weight x 100%. Enzyme soluble collagen yield = enzyme soluble collagen weight/fish scale weight × 100%. Keratin yield = keratin weight/fish scale weight × 100%. The above fish scale weights are dry weights.
The molecular weight distribution of the protein peptide is determined by adopting high-efficiency size exclusion chromatography according to the national standard of GB 31645 and 2018 for food safety.
Claims (10)
1. A process for extracting various nutrient components from fish scales is characterized by comprising the following steps:
(1) fish scale dehybridization protein
After cleaning fresh fish scales, soaking the fresh fish scales in a sodium chloride solution with the weight percentage concentration of 4-10% at 4-12 ℃ for 8-12 hours, taking out the fish scales, and rinsing the fish scales with clean water;
(2) degreasing of fish scales
Soaking the fish scales after the foreign protein removal in the step (1) in a sodium hydroxide solution with the weight percentage concentration of 0.4-0.8% at 4-12 ℃ for 4-6 hours, then taking out the fish scales, and rinsing the fish scales with clean water;
(3) decolouring fish scales
Soaking the degreased fish scales in the step (2) in a hydrogen peroxide solution with the weight percentage concentration of 1-2% for 4-8 hours at 4-12 ℃, taking out, and rinsing with clean water;
(4) fish scale deashing
Soaking the fish scales decolorized in the step (3) in an organic acid solution at 60-75 ℃ for 4-8 hours under the condition of stirring, standing after stirring, and separating a supernatant from a precipitate, wherein the supernatant is an organic acid calcium solution, and the precipitate is decalcified fish scales;
(5) organic acid calcium crystal
Freezing and crystallizing the supernatant obtained in the step (4) to obtain a crystal, namely the organic acid calcium;
(6) acid method and enzyme method for extracting gel
Adding the decalcified fish scales obtained in the step (4) into an organic acid solution, soaking at 4-8 ℃, and separating a dissolved solution from a precipitate after soaking is finished to obtain an acid-soluble collagen solution;
adding water into the precipitate to prepare a feed liquid, adding alkaline protease for enzymolysis to obtain an enzyme-soluble collagen solution, and separating the enzyme-soluble collagen solution to obtain a residue;
(7) collagen enzymolysis
Respectively carrying out enzymolysis on the acid-soluble collagen solution and the enzyme-soluble collagen solution obtained in the step (6) to obtain an acid-soluble collagen enzymolysis solution and an enzyme-soluble collagen enzymolysis solution;
(8) nanofiltration
Carrying out nanofiltration on the acid-soluble collagen enzymatic hydrolysate and the enzyme-soluble collagen enzymatic hydrolysate obtained in the step (7) respectively to obtain an acid-soluble collagen enzymatic concentrated solution and an enzyme-soluble collagen protease concentrated solution;
(9) sterilization
Respectively sterilizing the acid soluble collagen enzymolysis concentrated solution and the enzyme soluble collagenase concentrated solution obtained in the step (8);
(10) spray drying
Respectively carrying out spray drying on the acid-soluble collagen enzymolysis concentrated solution and the enzyme-soluble collagen protease concentrated solution obtained in the step (10) by using a spray dryer to respectively obtain acid-soluble collagen peptide powder and enzyme-soluble collagen peptide powder;
(11) alkaline process for the separation of keratin
And (3) mixing the residue obtained in the step (6) with a sodium hydroxide solution, reacting at 55-65 ℃ until the residue is completely dissolved, adding concentrated hydrochloric acid to adjust the pH value to the isoelectric point of keratin, standing, and separating out a precipitate, wherein the precipitate is the keratin.
2. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: in the step (4), the weight percentage concentration of the organic acid solution is 3-10%, and the organic acid is one or a combination of more of citric acid, malic acid and lactic acid;
in the step (4), the weight ratio of the decolored fish scales to the organic acid solution is 1: 8-1: 20;
in the step (4), stirring is carried out at a rotating speed of 30-50 revolutions per minute.
3. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: in the step (5), the freezing crystallization temperature is-8 to-12 ℃, and the crystallization time is 6 to 12 hours.
4. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: in the step (6), the weight percentage concentration of the organic acid solution is 5-10%, and the organic acid is one or a combination of more of acetic acid, citric acid, malic acid and lactic acid;
in the step (6), the weight ratio of the deashed fish scales to the organic acid solution is 1: 8-1: 20, and the soaking time is 18-24 hours.
5. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: in the step (6), the weight ratio of the precipitate to water is 1: 8-1: 20; adding alkaline protease in an amount of 1-3% by weight of the precipitate, and carrying out enzymolysis at 25-35 ℃ for 12-24 hours;
the activity unit of the alkaline protease used in the step (6) is 30-80 ten thousand units.
6. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: adding an acid-soluble collagen solution into a first enzymolysis tank, adding water to adjust the weight percentage concentration of the acid-soluble collagen to be 8-15%, using a ball mill to carry out homogenization, using sodium hydroxide to adjust the pH value to be 7.5-9, heating to 45-60 ℃, adding alkaline protease and neutral protease, wherein the weight of the alkaline protease is 1-3% of that of the acid-soluble collagen, and the weight of the neutral protease is 1-3% of that of the acid-soluble collagen; then carrying out enzymolysis for 4-8 hours under the condition of stirring, and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely an acid-soluble collagen enzymolysis liquid;
adding the enzyme-soluble collagen into a second enzymolysis tank, adding water to adjust the weight percentage concentration of the enzyme-soluble collagen to 8-15%, using a ball mill to carry out homogenate, using sodium hydroxide to adjust the pH value to 7.5-9, heating to 45-60 ℃, adding alkaline protease and neutral protease, wherein the weight of the alkaline protease is 1-3% of that of the enzyme-soluble collagen, and the weight of the neutral protease is 1-3% of that of the enzyme-soluble collagen; then carrying out enzymolysis for 4-8 hours under the condition of stirring, and filtering after the enzymolysis is finished to obtain an enzymolysis supernatant, namely an enzyme-soluble collagen enzymolysis liquid;
the activity unit of the alkaline protease used in the step (7) is 30-80 ten thousand units, and the activity unit of the neutral protease is 10-30 ten thousand units.
7. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: the aperture of the nanofiltration membrane used in the nanofiltration in the step (8) is 150-300 nanometers;
in the step (9), the sterilization mode is pasteurization, and the temperature is kept at 80-85 ℃ for 20-30 minutes;
in the step (10), the inlet air temperature of spray drying is 150-185 ℃, and the outlet air temperature is 60-85 ℃.
8. The process for extracting multiple nutrients from fish scales as claimed in claim 1, wherein the process comprises the steps of: in the step (11), the weight percentage concentration of the sodium hydroxide solution is 4-8%, and the weight of the sodium hydroxide solution is 10-20 times of that of the residue;
in the step (11), standing time is 30-60 minutes, and after standing, separating out precipitates by using a centrifugal machine, wherein the rotating speed of the centrifugal machine is 3000-5000 rpm, and the centrifugal time is 10-30 minutes.
9. The process of extracting nutrients from fish scales as claimed in claim 8, wherein: dissolving the precipitate separated in the step (11) by using a sodium hydroxide solution with the weight percentage concentration of 1-3%, and adjusting to be neutral by using hydrochloric acid to obtain a keratin solution; then carrying out nanofiltration on the obtained keratin solution to obtain a keratin concentrated solution; and sterilizing the keratin concentrated solution, and spray drying to obtain dry powdery keratin.
10. The process of extracting nutrients from fish scales as claimed in claim 8, wherein: and (3) freeze-drying the precipitate separated in the step (11) to obtain the dried keratin.
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