CN112493359A - Preparation method of fish scale collagen peptide glue powder - Google Patents
Preparation method of fish scale collagen peptide glue powder Download PDFInfo
- Publication number
- CN112493359A CN112493359A CN202011303644.XA CN202011303644A CN112493359A CN 112493359 A CN112493359 A CN 112493359A CN 202011303644 A CN202011303644 A CN 202011303644A CN 112493359 A CN112493359 A CN 112493359A
- Authority
- CN
- China
- Prior art keywords
- fish scale
- collagen peptide
- mass
- purified
- stirring
- 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
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 104
- 102000008186 Collagen Human genes 0.000 title claims abstract description 33
- 108010035532 Collagen Proteins 0.000 title claims abstract description 33
- 229920001436 collagen Polymers 0.000 title claims abstract description 33
- 239000000843 powder Substances 0.000 title claims abstract description 30
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 26
- 239000003292 glue Substances 0.000 title claims abstract description 18
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 239000004365 Protease Substances 0.000 claims abstract description 27
- 108091005804 Peptidases Proteins 0.000 claims abstract description 14
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 14
- 235000019419 proteases Nutrition 0.000 claims abstract description 14
- 108090000526 Papain Proteins 0.000 claims abstract description 13
- 108090000284 Pepsin A Proteins 0.000 claims abstract description 13
- 102000057297 Pepsin A Human genes 0.000 claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- 229940055729 papain Drugs 0.000 claims abstract description 13
- 235000019834 papain Nutrition 0.000 claims abstract description 13
- 229940111202 pepsin Drugs 0.000 claims abstract description 13
- 238000004537 pulping Methods 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 12
- 108091005508 Acid proteases Proteins 0.000 claims abstract description 11
- 235000011194 food seasoning agent Nutrition 0.000 claims abstract description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000009835 boiling Methods 0.000 claims abstract description 7
- 238000001694 spray drying Methods 0.000 claims abstract description 6
- 239000000203 mixture Substances 0.000 claims abstract description 5
- 239000000463 material Substances 0.000 claims description 98
- 238000003756 stirring Methods 0.000 claims description 30
- 239000000047 product Substances 0.000 claims description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 15
- 102000004190 Enzymes Human genes 0.000 claims description 14
- 108090000790 Enzymes Proteins 0.000 claims description 14
- 229940088598 enzyme Drugs 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 238000007873 sieving Methods 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 238000004140 cleaning Methods 0.000 claims description 8
- 229920000159 gelatin Polymers 0.000 claims description 8
- 235000019322 gelatine Nutrition 0.000 claims description 8
- 239000012535 impurity Substances 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 6
- 238000000746 purification Methods 0.000 claims description 6
- 241000252230 Ctenopharyngodon idella Species 0.000 claims description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims description 4
- 108010010803 Gelatin Proteins 0.000 claims description 4
- 239000001828 Gelatine Substances 0.000 claims description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 239000008273 gelatin Substances 0.000 claims description 4
- 235000011852 gelatine desserts Nutrition 0.000 claims description 4
- 239000002002 slurry Substances 0.000 claims description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims description 2
- 239000004375 Dextrin Substances 0.000 claims description 2
- 229920001353 Dextrin Polymers 0.000 claims description 2
- 229930091371 Fructose Natural products 0.000 claims description 2
- 239000005715 Fructose Substances 0.000 claims description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 2
- 241000252234 Hypophthalmichthys nobilis Species 0.000 claims description 2
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 claims description 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 230000002378 acidificating effect Effects 0.000 claims description 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
- 235000019425 dextrin Nutrition 0.000 claims description 2
- 239000013505 freshwater Substances 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 239000000796 flavoring agent Substances 0.000 abstract description 8
- 235000019634 flavors Nutrition 0.000 abstract description 8
- 239000000126 substance Substances 0.000 abstract description 4
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract description 2
- 230000015556 catabolic process Effects 0.000 abstract description 2
- 238000006731 degradation reaction Methods 0.000 abstract description 2
- 238000004090 dissolution Methods 0.000 abstract description 2
- 229920002521 macromolecule Polymers 0.000 abstract description 2
- 150000005846 sugar alcohols Chemical class 0.000 abstract 1
- 235000013305 food Nutrition 0.000 description 8
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 4
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000001502 supplementing effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 108010005094 Advanced Glycation End Products Proteins 0.000 description 1
- 239000004475 Arginine Substances 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
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 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
- 229940079593 drug Drugs 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 230000032050 esterification Effects 0.000 description 1
- 238000005886 esterification reaction Methods 0.000 description 1
- 108010007119 flavourzyme Proteins 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 235000021190 leftovers Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 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 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
- A23J3/342—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of collagen; of gelatin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/20—Synthetic spices, flavouring agents or condiments
- A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
- A23L27/215—Synthetic spices, flavouring agents or condiments containing amino acids heated in the presence of reducing sugars, e.g. Maillard's non-enzymatic browning
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/40—Shaping or working of foodstuffs characterised by the products free-flowing powder or instant powder, i.e. powder which is reconstituted rapidly when liquid is added
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Nutrition Science (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a preparation method of fish scale collagen peptide glue powder, which mainly comprises the following steps: the fish scale raw material is taken, crushed, pulped, separated and decontaminated after being purified, pepsin, acid protease, papain and compound protease are sequentially added for multi-stage enzymolysis, edible sugar and edible alcohol are sequentially added for seasoning and decocting, and the mixture is prepared into powder which is convenient to take after spray drying. The invention comprehensively utilizes the technical measures of pulping, multi-stage enzymolysis and the like, greatly promotes the degradation of macromolecules of the collagen, improves the dissolution efficiency of the product, not only improves the utilization rate of raw materials, but also improves the nutritional value of the product; the seasoning and boiling enrich the flavor substances and the active ingredients, improve the taste and further improve the product value.
Description
Technical Field
The invention relates to the technical field of food nutrition and body building products, in particular to a preparation method of fish scale collagen peptide gelatine powder.
Background
Collagen is a high molecular protein rich in human and animal bodies and widely distributed in connective tissues such as skin, tendon, fish scales and the like; contains special amino acid types and proportions, and has wide application in the fields of food, cosmetics, medicines, biomedical materials and the like.
China is a large country for aquatic product production and consumption, and a large amount of fish skin, fish scale and other leftovers generated in the processing process of aquatic product fish are mostly treated as wastes at present, so that resources are wasted, and environmental pollution is caused. In recent years, research on fish scale components and functions thereof shows that the content of protein in the dry substance of fish scales is as high as 70-80%, and the dry substance of fish scales is rich in essential amino acids and functional amino acids which are beneficial to human bodies, such as proline, hydroxyproline, glycine, arginine, lysine and the like. Compared with mammalian collagen such as pigskin, cow leather and the like, the fish scale and fish skin collagen can be developed into food and cosmetics to avoid potential risks of protein homology in human body application.
Therefore, the development and comprehensive utilization of fish scale products are highly valued at home and abroad, wherein the fish scale collagen peptide is one of important products. However, the prior production process of fish scale collagen peptide has the defects of low utilization rate of raw materials, single product component, low product yield and the like.
Disclosure of Invention
The invention aims to solve the technical problem of providing a preparation method of fish scale collagen peptide gelatine powder convenient for eating, so as to improve the utilization rate of raw materials and the overall production efficiency, improve the appearance and flavor of finished products and improve the nutritional value of the finished products.
In order to solve the technical problems, the invention adopts the following technical scheme:
a preparation method of fish scale collagen peptide gelatine powder is developed, and comprises the following steps:
(1) and (3) purification treatment: washing a fish scale raw material to remove silt and other impurities to obtain a purified fish scale material;
(2) crushing and pulping: crushing and pulping the purified fish scale material into a slurry material;
(3) separating and removing impurities: adding 1-1.5% citric acid solution (to remove hydroxyapatite in fish scales) 3-5 times of the mass of the fish scale purification material, stirring and soaking at normal temperature for 6-8 h, sieving with a 100-120 mesh sieve, collecting oversize products, and cleaning with clear water for 1-3 times;
(4) multi-stage enzymolysis
Adding water into the materials obtained in the previous step according to the mass which is 3-5 times of that of the purified fish scale materials, heating to 35-38 ℃, adjusting the pH value to 2-3 by using dilute acid, adding pepsin, keeping the temperature, stirring and reacting for 1.5-2.5 h, wherein the adding amount of the pepsin is 0.02-0.05% of the mass of the purified fish scale materials;
heating the enzymolysis material obtained in the previous step to 40-55 ℃, adding acid protease into the enzymolysis material, and carrying out heat preservation reaction for 2-4.5 hours under the stirring condition, wherein the adding amount of the acid protease is 0.5-1.0% of the mass of the purified fish scale material;
adding papain into the enzymolysis material obtained in the previous step, stirring and reacting for 3-6 hours at 40-55 ℃, wherein the adding amount of the papain is 0.3-0.8% of the mass of the purified fish scale material;
finally, adding compound protease into the enzymolysis material obtained in the previous step, stirring and reacting for 1-3.0 hours at 40-55 ℃, wherein the adding amount of the compound protease is 0.4-1.0% of the mass of the purified fish scale material;
(5) seasoning and boiling: sieving the enzymolysis material obtained in the previous step by a sieve of 80-120 meshes, washing the oversize material for 1-2 times by hot water at 40-55 ℃, combining the filtrates, heating to 80-95 ℃, adding reducing sugar into the mixture according to 0.8-1.2% of the mass of the purified fish scale material, stirring, keeping the temperature, decocting for 1.0-1.5 h, adding edible alcohol into the mixture according to 0.3-0.5% of the mass of the purified fish scale material, and continuing to keep the temperature and decocting for 1-1.5 h;
(6) drying to prepare powder: and (4) spray drying the feed liquid obtained in the previous step to obtain the fish scale collagen peptide glue powder.
Further, the specification of the pepsin is 1: 3000.
Further, the papain is 60-70 ten thousand units/g of active food-grade enzyme preparation.
Further, in the step (4), the acidic protease is 5-15 ten thousand units/g of active food-grade enzyme preparation.
Further, in the step (4), the activity of the compound protease is 1.5 AU/g.
Further, in the step (5), the edible sugar is at least one of glucose, sucrose, maltose, lactose, fructose, xylose, arabinose, dextrin, mannose and rhamnose.
The fish scale is freshwater fish scale, such as grass carp scale, silver carp scale, crucian scale, etc.
Compared with the prior art, the invention has the main beneficial technical effects that:
1. the invention comprehensively utilizes the technological measures of pulping, multi-stage enzymolysis and the like, effectively improves the utilization rate of raw materials (by about 15-20 percent, has high production efficiency and reduces the production cost, and the components can be directly absorbed and utilized by human bodies.
2. In the preparation process, the degradation of macromolecules of the collagen is greatly promoted through multi-stage enzymolysis, more small peptides are generated, the dissolution efficiency of the product is improved, the utilization rate of raw materials is improved, and the nutritive value of the finished product is greatly improved and enriched.
3. According to the invention, edible sugar (forming a Maillard product, not only generating functional components but also improving the flavor) and edible alcohol (removing peculiar smell, promoting esterification and improving the flavor) are sequentially added in the seasoning and boiling stage, so that the taste phase is improved, flavor substances and active ingredients are increased, and the product value is further improved.
4. The product of the invention is powder, is convenient to eat and is more convenient to package and transport.
Detailed Description
The following examples are given to illustrate specific embodiments of the present invention, but are not intended to limit the scope of the present invention in any way.
The apparatuses referred to in the following examples are conventional apparatuses unless otherwise specified; the related food raw materials are all conventional products sold in the market if not specified; the test detection methods are conventional methods unless otherwise specified.
Embodiment 1 a method for preparing fish scale collagen peptide gelatin powder, which mainly comprises the following steps:
(1) and (3) purification treatment: repeatedly washing the grass carp scale raw material in a cleaning tank by using clear water until no silt and sundries exist, and obtaining the purified fish scale material.
(2) Pulping: and crushing and pulping the purified fish scale material into pulp with the granularity less than 2 mm.
(3) Separating and removing impurities: adding 1% citric acid solution 4 times of the purified fish scale material, stirring at room temperature, soaking for 7 hr, sieving with 120 mesh sieve, collecting oversize, and cleaning with clear water for 2 times.
(4) Multi-stage enzymolysis:
adding water into the materials obtained in the previous step according to 4 times of the mass of the purified fish scale materials, heating to 38 ℃, adjusting the pH value to 2.5 by using 6 mol/L dilute hydrochloric acid, adding pepsin (specification is 1: 3000), keeping the temperature, stirring and reacting for 2.0 h, wherein the adding amount of the pepsin is 0.03 percent of the mass of the purified fish scale materials;
heating the enzymolysis material obtained in the previous step to 55 ℃, adding acid protease (8 ten thousand units/g active food grade enzyme preparation), and reacting for 3.5 hours under stirring at a heat preservation condition, wherein the adding amount of the acid protease is 0.8 percent of the mass of the purified fish scale material;
adding papain (60 ten thousand units/g active food grade enzyme preparation) into the enzymolysis material obtained in the previous step, and stirring and reacting at 55 ℃ for 4.5 hours, wherein the adding amount of the papain is 0.5 percent of the mass of the purified fish scale material;
and finally, adding compound protease (Protamex, Novit company, with the enzyme activity of 1.5 AU/g) into the enzymolysis material obtained in the last step, stirring and reacting for 3.0 hours at the temperature of 55 ℃, wherein the adding amount of the compound protease is 0.5 percent of the mass of the purified fish scale material.
(5) Seasoning and boiling: sieving the enzymolysis material obtained in the previous step with a 100-mesh sieve, washing the oversize material with 55 ℃ hot water which is 0.8 time of the mass of the purified fish scale material for 1 time, combining filtrates, heating to 90 ℃, adding glucose into the purified fish scale material according to 1% of the mass of the purified fish scale material, stirring, keeping the temperature, decocting for 1.5 hours, adding edible alcohol into the purified fish scale material according to 0.4% of the mass of the purified fish scale material, and continuously keeping the temperature and decocting for 1.5 hours; after decocting, the volume of the materials is kept stable by supplementing hot water with the temperature of 85 ℃.
(6) Drying to prepare powder: spray drying the feed liquid obtained in the previous step to obtain the fish scale collagen peptide glue powder (the water content is 6%).
The results of the detection, evaluation and analysis of the obtained product are as follows:
the product yield is 22.8%; protein content 81.7%; wherein the content of important amino acid is as follows: proline 7.15%; 15.82 percent of glycine; the flavor and taste of the finished product are characterized by collagen glue.
Embodiment 2 a method for preparing fish scale collagen peptide gelatin powder, which mainly comprises the following steps:
(1) and (3) purification treatment: repeatedly washing the grass carp scale raw material in a cleaning tank by using clear water until no silt and sundries exist, and obtaining the purified fish scale material.
(2) Pulping: and crushing and pulping the purified fish scale material into pulp with the granularity less than 2 mm.
(3) Separating and removing impurities: adding 1% citric acid solution 5 times of the purified fish scale material, stirring at room temperature, soaking for 7 hr, sieving with 120 mesh sieve, collecting oversize, and cleaning with clear water for 2 times.
(4) Multi-stage enzymolysis:
adding water into the materials obtained in the previous step according to 4 times of the mass of the purified fish scale materials, heating to 36 ℃, adjusting the pH value to 3.0 by using 6 mol/L dilute hydrochloric acid, adding pepsin (specification is 1: 3000), preserving heat, stirring and reacting for 2.0 h, wherein the adding amount of the pepsin is 0.03 percent of the mass of the purified fish scale materials;
heating the enzymolysis material obtained in the previous step to 50 ℃, adding acid protease (8 ten thousand units/g active food grade enzyme preparation), and carrying out heat preservation reaction for 2.5 hours under the stirring condition, wherein the adding amount of the acid protease is 1.0 percent of the mass of the purified fish scale material;
adding papain (60 ten thousand units/g active food-grade enzyme preparation) into the enzymolysis material obtained in the previous step, and stirring and reacting at 50 ℃ for 3.5 hours, wherein the adding amount of the papain is 0.8 percent of the mass of the purified fish scale material;
and finally, adding compound protease (Protamex, Novit company, with the enzyme activity of 1.5 AU/g) into the enzymolysis material obtained in the last step, stirring and reacting for 2.0 hours at 50 ℃, wherein the adding amount of the compound protease is 0.7 percent of the mass of the purified fish scale material.
(5) Seasoning and boiling: sieving the enzymolysis material obtained in the previous step with a 100-mesh sieve, washing the oversize material with hot water at 55 ℃ which is 0.7 time of the mass of the purified fish scale material for 1 time, combining filtrates, heating to 95 ℃, adding glucose into the purified fish scale material according to 1% of the mass of the purified fish scale material, stirring, keeping the temperature, decocting for 1.5 hours, adding edible alcohol into the purified fish scale material according to 0.4% of the mass of the purified fish scale material, and continuing to keep the temperature and decocting for 1.5 hours; after decocting, the volume of the materials is kept stable by supplementing hot water with the temperature of 85 ℃.
(6) Drying to prepare powder: spray drying the feed liquid obtained in the previous step to obtain the fish scale collagen peptide glue powder (the water content is 6%).
The results of the detection, evaluation and analysis of the obtained product are as follows:
the product yield is 23.1%; protein content 82.6%; wherein the content of important amino acid is as follows: proline 6.93%; 16.03 percent of glycine; the flavor and taste of the finished product are characterized by collagen glue.
Embodiment 3 a method for preparing fish scale collagen peptide gelatin powder, which mainly comprises the following steps:
(1) and (3) purification treatment: repeatedly washing the grass carp scale raw material in a cleaning tank by using clear water until no silt and sundries exist, and obtaining the purified fish scale material.
(2) Pulping: and crushing and pulping the purified fish scale material into pulp with the granularity less than 2 mm.
(3) Separating and removing impurities: adding 1% citric acid solution 3 times of the purified fish scale material, stirring at room temperature, soaking for 7 hr, sieving with 120 mesh sieve, collecting oversize, and cleaning with clear water for 2 times.
(4) Multi-stage enzymolysis:
adding water into the materials obtained in the previous step according to 4 times of the mass of the purified fish scale materials, heating to 35 ℃, adjusting the pH value to 2.0 by using 6 mol/L dilute hydrochloric acid, adding pepsin (specification is 1: 3000), keeping the temperature, stirring and reacting for 2.0 h, wherein the adding amount of the pepsin is 0.03 percent of the mass of the purified fish scale materials;
heating the enzymolysis material obtained in the previous step to 45 ℃, adding acid protease (8 ten thousand units/g active food grade enzyme preparation), and carrying out heat preservation reaction for 4.0h under the stirring condition, wherein the adding amount of the acid protease is 0.7 percent of the mass of the purified fish scale material;
adding papain (60 ten thousand units/g active food-grade enzyme preparation) into the enzymolysis material obtained in the previous step, and stirring and reacting at 45 ℃ for 4.0h, wherein the adding amount of the papain is 0.6% of the mass of the purified fish scale material;
and finally, adding compound protease (Protamex, Novit company, with the enzyme activity of 1.5 AU/g) into the enzymolysis material obtained in the last step, stirring and reacting for 1.5 hours at 45 ℃, wherein the adding amount of the compound flavourzyme is 0.8 percent of the mass of the purified fish scale material.
(5) Seasoning and boiling: sieving the enzymolysis material obtained in the previous step with a 100-mesh sieve, washing the oversize material with 55 ℃ hot water which is 0.5 time of the mass of the purified fish scale material for 1 time, combining filtrates, heating to 85 ℃, adding glucose into the purified fish scale material according to 1% of the mass of the purified fish scale material, stirring, keeping the temperature, decocting for 1.5 hours, adding edible alcohol into the purified fish scale material according to 0.4% of the mass of the purified fish scale material, and continuously keeping the temperature and decocting for 1.5 hours; after decocting, the volume of the materials is kept stable by supplementing hot water with the temperature of 85 ℃.
(6) Drying to prepare powder: spray drying the feed liquid obtained in the previous step to obtain the fish scale collagen peptide glue powder (the water content is 6%).
The results of the detection, evaluation and analysis of the obtained product are as follows:
the product yield is 23.6%; protein content 82.3%; wherein the content of important amino acid is as follows: proline 7.03%; 15.96 percent of glycine; the flavor and taste of the finished product are characterized by collagen glue.
The present invention is described in detail with reference to the examples above; however, it will be understood by those skilled in the art that various changes in the specific components and process parameters of the above embodiments may be made without departing from the spirit of the invention, or equivalents thereof may be substituted for elements thereof, thereby forming a plurality of specific embodiments, which are common variations of the invention and will not be described in detail herein.
Claims (9)
1. A preparation method of fish scale collagen peptide gelatine powder is characterized by comprising the following steps:
(1) purification treatment
Washing a fish scale raw material to remove silt and impurities to obtain a purified fish scale material;
(2) crushing and pulping
Crushing and pulping the purified fish scale material into a slurry material;
(3) separating and removing impurities
Adding a citric acid solution with the concentration of 1.0-1.5% according to 3-5 times of the mass of the purified fish scale materials, stirring and soaking at normal temperature for 6-8 h, sieving with a 100-120 mesh sieve, collecting oversize products, and cleaning with clear water for 1-3 times;
(4) multi-stage enzymolysis
Adding water into the materials obtained in the previous step according to the mass which is 3-5 times of that of the purified fish scale materials, heating to 35-38 ℃, adjusting the pH value to 2-3 by using dilute acid, adding pepsin, keeping the temperature, stirring and reacting for 1.5-2.5 h, wherein the adding amount of the pepsin is 0.02-0.05% of the mass of the purified fish scale materials;
heating the enzymolysis material obtained in the previous step to 40-55 ℃, adding acid protease into the enzymolysis material, and carrying out heat preservation reaction for 2-4.5 hours under the stirring condition, wherein the adding amount of the acid protease is 0.5-1.0% of the mass of the purified fish scale material;
adding papain into the enzymolysis material obtained in the previous step, stirring and reacting for 3-6 hours at 40-55 ℃, wherein the adding amount of the papain is 0.3-0.8% of the mass of the purified fish scale material;
finally, adding compound protease into the enzymolysis material obtained in the previous step, stirring and reacting for 1-3.0 hours at 40-55 ℃, wherein the adding amount of the compound protease is 0.4-1.0% of the mass of the purified fish scale material;
(5) seasoning and boiling
Sieving the enzymolysis material obtained in the previous step by a sieve of 80-120 meshes, washing the oversize material for 1-2 times by hot water at 40-55 ℃, combining the filtrates, heating to 80-95 ℃, adding reducing sugar into the mixture according to 0.8-1.2% of the mass of the purified fish scale material, stirring, keeping the temperature, decocting for 1.0-1.5 h, adding edible alcohol into the mixture according to 0.3-0.5% of the mass of the purified fish scale material, and continuing to keep the temperature and decocting for 1-1.5 h;
(6) drying to obtain powder
And (4) spray drying the feed liquid obtained in the previous step to obtain the fish scale collagen peptide glue powder.
2. The method for preparing the fish scale collagen peptide glue powder according to claim 1, wherein in the step (2), the purified fish scales are crushed and pulped into a slurry material with the granularity less than or equal to 2 mm.
3. The method for preparing donkey-hide fish scale collagen peptide gelatin powder according to claim 1, wherein in the step (4), the specification of pepsin is 1: 3000.
4. The method for preparing the fish scale collagen peptide glue powder according to claim 1, wherein in the step (4), the papain is 60-70 ten thousand units/g of active food-grade enzyme preparation.
5. The method for preparing the fish scale collagen peptide glue powder according to claim 1, wherein in the step (4), the acidic protease is 5-15 ten thousand units/g of active food-grade enzyme preparation.
6. The method for preparing fish scale collagen peptide glue powder according to claim 1, wherein in the step (4), the enzyme activity of the compound protease is 1.5 AU/g.
7. The method for preparing fish scale collagen peptide glue powder according to claim 1, wherein in the step (5), the edible sugar is at least one of glucose, sucrose, maltose, lactose, fructose, xylose, arabinose, dextrin, mannose and rhamnose.
8. The method for preparing the fish scale collagen peptide glue powder of claim 1, wherein the fish scales are scales of freshwater fish.
9. The method for preparing the fish scale collagen peptide glue powder according to claim 8, wherein the fish scale is at least one of grass carp scales, silver carp scales and crucian scales.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011303644.XA CN112493359A (en) | 2020-11-19 | 2020-11-19 | Preparation method of fish scale collagen peptide glue powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011303644.XA CN112493359A (en) | 2020-11-19 | 2020-11-19 | Preparation method of fish scale collagen peptide glue powder |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112493359A true CN112493359A (en) | 2021-03-16 |
Family
ID=74958862
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011303644.XA Pending CN112493359A (en) | 2020-11-19 | 2020-11-19 | Preparation method of fish scale collagen peptide glue powder |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112493359A (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007159557A (en) * | 2005-11-21 | 2007-06-28 | Meiji Seika Kaisha Ltd | Method for removing collagen smell, and applied food and drink of the same |
| JP2008206470A (en) * | 2007-02-27 | 2008-09-11 | Shinhui Bio-Tech Co Ltd | Method for producing collagen peptide of fish scale origin |
| CN102154424A (en) * | 2011-01-14 | 2011-08-17 | 国家海洋局第三海洋研究所 | Process for preparing micro-molecular fish scale collagen peptides |
| CN102172393A (en) * | 2010-12-17 | 2011-09-07 | 中国海洋大学 | Calcium supplement of fish scale protein peptide calcium chelate and preparation method thereof |
| CN102839207A (en) * | 2012-09-17 | 2012-12-26 | 温州大学 | Method for preparing collagen peptide from fish scales |
| CN105039483A (en) * | 2015-09-21 | 2015-11-11 | 广东海洋大学 | Method for preparing fishskin fish-scale collagen protein |
| JP2017051176A (en) * | 2015-05-19 | 2017-03-16 | 株式会社和冠 | Sweetener composition and method for producing the same |
| CN111748599A (en) * | 2020-07-28 | 2020-10-09 | 卫青健康科技(苏州)有限公司 | Preparation method of fish scale collagen peptide |
-
2020
- 2020-11-19 CN CN202011303644.XA patent/CN112493359A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007159557A (en) * | 2005-11-21 | 2007-06-28 | Meiji Seika Kaisha Ltd | Method for removing collagen smell, and applied food and drink of the same |
| JP2008206470A (en) * | 2007-02-27 | 2008-09-11 | Shinhui Bio-Tech Co Ltd | Method for producing collagen peptide of fish scale origin |
| CN102172393A (en) * | 2010-12-17 | 2011-09-07 | 中国海洋大学 | Calcium supplement of fish scale protein peptide calcium chelate and preparation method thereof |
| CN102154424A (en) * | 2011-01-14 | 2011-08-17 | 国家海洋局第三海洋研究所 | Process for preparing micro-molecular fish scale collagen peptides |
| CN102839207A (en) * | 2012-09-17 | 2012-12-26 | 温州大学 | Method for preparing collagen peptide from fish scales |
| JP2017051176A (en) * | 2015-05-19 | 2017-03-16 | 株式会社和冠 | Sweetener composition and method for producing the same |
| CN105039483A (en) * | 2015-09-21 | 2015-11-11 | 广东海洋大学 | Method for preparing fishskin fish-scale collagen protein |
| CN111748599A (en) * | 2020-07-28 | 2020-10-09 | 卫青健康科技(苏州)有限公司 | Preparation method of fish scale collagen peptide |
Non-Patent Citations (1)
| Title |
|---|
| 陈露: "鲤鱼鱼鳞胶原蛋白肽的制备工艺和分析" * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103173511A (en) | Method for industrially producing wheat glutamine peptide | |
| CN101033481A (en) | Preparing process of extracting bone collagen from pig, cattle and sheep bone | |
| CN104095141A (en) | Method for coproducing collagen activity peptides and flavoured base materials with chicken bones | |
| CN108477619A (en) | A kind of preparation method of compound amino acid chelate calcium | |
| CN105147727A (en) | Preparation method of Qinghai-Tibet plateau yak skin colla corii asini | |
| CN108771244A (en) | The preparation method of the oligomeric peptide extract of abalone, method, abalone oligopeptide and its application for preparing abalone oligopeptide and abalone powder | |
| CN108752466A (en) | A kind of chelated calcium preparation method of tuna bone collagen peptide | |
| CN108935912B (en) | Fish meat protein peptide with DPP-IV inhibition and anti-fatigue functions and preparation method thereof | |
| CN109007746A (en) | A kind of preparation method of sturgeon head seasoning | |
| CN108727485A (en) | A kind of preparation method of food-grade hydrolysis of keratin | |
| CN106566861B (en) | Preparation method of black-bone chicken oligopeptide mixed powder with high F value | |
| CN101649341A (en) | Method for extracting protein peptide from membranes of fowl eggshells | |
| CN104531663A (en) | Immobilized enzyme hydrolysis method for preparing high F value oligopeptide by using minced squid meat | |
| CN103966291A (en) | Yeast peptone | |
| CN112493359A (en) | Preparation method of fish scale collagen peptide glue powder | |
| KR101115510B1 (en) | The extracting method of constituents of silkworm and so on, characterized by liquidified extraction by enzyme treatment at low temperature | |
| CN106616841A (en) | Preparation method of Corbicula fluminea seasoning | |
| US20040248239A1 (en) | Method for producing amino acid components by enzymatic hydrolysis of fish egg skin | |
| CN115349623A (en) | Coconut flavor base material and preparation method and application thereof | |
| CN108840903A (en) | A kind of extracting method of rice protein powder | |
| CN1276723C (en) | Water soluble royal jelly and its dry powder and method for preparing them | |
| CN107760747A (en) | A kind of industrialized production earthworm protein active peptide and preparation method | |
| CN101449737A (en) | Preparation method of scallop pure skirt amino acid nutrient powder | |
| CN112471448A (en) | Preparation method of fish scale collagen peptide iron zinc chelate | |
| AU2020101636A4 (en) | A preparation method and product of broad bean water extract feed and its 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 |