CN110590938A - Preparation method of collagen peptide-zinc chelate - Google Patents
Preparation method of collagen peptide-zinc chelate Download PDFInfo
- Publication number
- CN110590938A CN110590938A CN201910783971.0A CN201910783971A CN110590938A CN 110590938 A CN110590938 A CN 110590938A CN 201910783971 A CN201910783971 A CN 201910783971A CN 110590938 A CN110590938 A CN 110590938A
- Authority
- CN
- China
- Prior art keywords
- collagen
- collagen peptide
- zinc
- peptide
- zinc chelate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000008186 Collagen Human genes 0.000 title claims abstract description 152
- 108010035532 Collagen Proteins 0.000 title claims abstract description 152
- 229920001436 collagen Polymers 0.000 title claims abstract description 152
- 229910052725 zinc Inorganic materials 0.000 title claims abstract description 55
- 239000011701 zinc Substances 0.000 title claims abstract description 55
- 239000013522 chelant Substances 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 68
- 239000002994 raw material Substances 0.000 claims abstract description 36
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 18
- 235000015170 shellfish Nutrition 0.000 claims abstract description 12
- 238000000605 extraction Methods 0.000 claims abstract description 11
- 108091005804 Peptidases Proteins 0.000 claims abstract description 4
- 239000004365 Protease Substances 0.000 claims abstract description 4
- 238000004925 denaturation Methods 0.000 claims abstract description 4
- 230000036425 denaturation Effects 0.000 claims abstract description 4
- 229910052751 metal Inorganic materials 0.000 claims abstract description 4
- 239000002184 metal Substances 0.000 claims abstract description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 50
- 239000000243 solution Substances 0.000 claims description 46
- 239000012153 distilled water Substances 0.000 claims description 38
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 32
- 239000002244 precipitate Substances 0.000 claims description 20
- 238000004140 cleaning Methods 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 13
- 238000004108 freeze drying Methods 0.000 claims description 12
- 239000012266 salt solution Substances 0.000 claims description 12
- 150000003751 zinc Chemical class 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 102000004190 Enzymes Human genes 0.000 claims description 8
- 108090000790 Enzymes Proteins 0.000 claims description 8
- 102000004142 Trypsin Human genes 0.000 claims description 8
- 108090000631 Trypsin Proteins 0.000 claims description 8
- 230000009920 chelation Effects 0.000 claims description 8
- 229940088598 enzyme Drugs 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 239000012588 trypsin Substances 0.000 claims description 8
- 102000057297 Pepsin A Human genes 0.000 claims description 7
- 108090000284 Pepsin A Proteins 0.000 claims description 7
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 229940111202 pepsin Drugs 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 239000012535 impurity Substances 0.000 claims description 6
- 230000010355 oscillation Effects 0.000 claims description 6
- 238000002791 soaking Methods 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 230000007935 neutral effect Effects 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 230000003301 hydrolyzing effect Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 18
- 241001465754 Metazoa Species 0.000 abstract description 7
- 201000010099 disease Diseases 0.000 abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 6
- 208000007212 Foot-and-Mouth Disease Diseases 0.000 abstract description 4
- 241000710198 Foot-and-mouth disease virus Species 0.000 abstract description 4
- 235000013376 functional food Nutrition 0.000 abstract description 4
- 230000002195 synergetic effect Effects 0.000 abstract description 4
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract description 3
- 230000003262 anti-osteoporosis Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 13
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- 241000490567 Pinctada Species 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 5
- 230000011164 ossification Effects 0.000 description 5
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 5
- 229960001763 zinc sulfate Drugs 0.000 description 5
- 229910000368 zinc sulfate Inorganic materials 0.000 description 5
- 241000283690 Bos taurus Species 0.000 description 4
- 208000001132 Osteoporosis Diseases 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 210000000988 bone and bone Anatomy 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 150000004697 chelate complex Chemical class 0.000 description 4
- 238000005520 cutting process Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 210000000963 osteoblast Anatomy 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000000415 inactivating effect Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 238000000967 suction filtration Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 229940038879 chelated zinc Drugs 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000009849 deactivation Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 201000008482 osteoarthritis Diseases 0.000 description 2
- 239000000902 placebo Substances 0.000 description 2
- 229940068196 placebo Drugs 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 241000272517 Anseriformes Species 0.000 description 1
- 208000006386 Bone Resorption Diseases 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 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
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000002979 Influenza in Birds Diseases 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241000276707 Tilapia Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 206010048259 Zinc deficiency Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 206010064097 avian influenza Diseases 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 210000002805 bone matrix Anatomy 0.000 description 1
- 230000024279 bone resorption Effects 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001934 delay 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
- 230000002875 effect on osteoarthritis Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229930182833 estradiol Natural products 0.000 description 1
- 229960005309 estradiol Drugs 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-N hydroperoxyl Chemical compound O[O] OUUQCZGPVNCOIJ-UHFFFAOYSA-N 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 230000004072 osteoblast differentiation Effects 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- AVPCPPOOQICIRJ-UHFFFAOYSA-L sodium glycerol 2-phosphate Chemical compound [Na+].[Na+].OCC(CO)OP([O-])([O-])=O AVPCPPOOQICIRJ-UHFFFAOYSA-L 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000001519 tissue Anatomy 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
- 230000001228 trophic effect Effects 0.000 description 1
- 238000005406 washing Methods 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]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a preparation method of a collagen peptide-zinc chelate, which takes fresh shellfish as a raw material, obtains water-soluble collagen of a shellfish mantle after denaturation by adopting a heat extraction method, then hydrolyzes the water-soluble collagen by using protease to prepare collagen peptide, and chelates the collagen peptide with metal zinc ions to prepare the collagen peptide-zinc chelate. The method takes the processing waste of marine organisms as the raw material, and has the advantages of simple and convenient acquisition way, low cost, high chelating rate and capability of avoiding the risks of mad cow disease, foot and mouth disease and the like possibly occurring in the collagen of the terrestrial animals. Meanwhile, compared with zinc ions, the peptide-zinc chelate chelated by the collagen peptide and the zinc ions has small toxic and side effects, can exert the synergistic effect of the collagen peptide and the zinc ions, and can provide safe and well-effective anti-osteoporosis functional food for consumers.
Description
Technical Field
The invention belongs to the technical field of biochemistry, and relates to a preparation method of a collagen peptide-zinc chelate.
Background
Bone homeostasis is achieved by the co-regulation of osteoblast mediated bone formation and osteoclast mediated bone resorption. Environmental, age, genetic factors and the like can cause imbalance of bone homeostasis and degradation of bone microstructure, and then diseases such as osteoporosis and osteoarthritis are caused. At present, the treatment of osteoporosis is mainly focused on methods using estradiol, calcitonin, diphosphonates and the like, and the medicaments can generate a large number of side effects after being taken for a long time. Therefore, the search for safe and effective foods and base materials which can be taken for a long time based on the medical and food homologous theory has the strategic significance of discovering and guiding the mainstream functional foods in the 21 st century.
Collagen is widely distributed in human and animal tissues, and at least 27 types of collagen have been found to date. Collagen has potential therapeutic efficacy on inflammation, tumor, osteoporosis, etc. and is considered as a safe food. There is a literature that indicates that the biological activity of collagen peptides is not only related to their molecular weight but also to the source and type of collagen. Research shows that the collagen peptide of terrestrial animals including cattle, pigs, geese and the like has potential therapeutic effect on osteoporosis and osteoarthritis. However, in recent years, diseases such as mad cow disease, foot and mouth disease, avian influenza and the like, which are continuously outbreaked, cause great concern to the collagen of the terrestrial animals, and the collagen of the aquatic animals has attracted attention in recent years because of high quality, safety and wide application range.
Zinc is an essential trophic factor in bone formation and growth. In recent years, there is increasing evidence that zinc deficiency inhibits mineralization of bone matrix and delays bone formation. Although inorganic zinc can be taken to supplement the deficiency of zinc, inorganic zinc has the characteristics of poor stability and absorption effect, strong toxicity to human bodies, unsuitability for long-term taking and the like. The peptide-zinc chelate not only can further improve the efficacy of the collagen peptide and the absorptivity and stability of the zinc element, but also can achieve the purpose of exerting a synergistic effect. However, the chelating effect of the collagen peptide obtained by using the common type I collagen as the raw material is not ideal when chelating zinc, and the yield of the collagen peptide chelated zinc is low. Therefore, it is an urgent problem to grasp a preparation technology of a chelate product combining collagen peptide and zinc, to improve the production efficiency of collagen peptide chelated zinc, and to satisfy the consumer demand.
Disclosure of Invention
The invention aims to provide a preparation method of collagen peptide-zinc chelate, which is used for preparing V-type shellfish collagen peptide-zinc chelate to promote bone formation.
The technical scheme adopted by the invention is that a preparation method of collagen peptide-zinc chelate is characterized in that fresh shellfish is used as a raw material, a thermal extraction method is adopted to obtain water-soluble collagen of a shellfish mantle after denaturation, then protease is used to hydrolyze the water-soluble collagen to prepare collagen peptide, and the collagen peptide is chelated with metal zinc ions to prepare the collagen peptide-zinc chelate.
The invention is also characterized in that the method,
the method is implemented by the following steps:
step 1, cleaning a fresh shellfish mantle with distilled water, removing impurities, and processing into small raw materials;
step 2, removing foreign proteins in the raw materials processed in the step 1 to obtain collagen, and preparing denatured collagen by adopting a thermal extraction method;
step 3, carrying out enzymolysis on the denatured collagen obtained in the step 2 by adopting pepsin or trypsin to obtain collagen peptide;
and 4, carrying out chelation reaction on the collagen peptide obtained in the step 3 and a zinc salt solution, separating and drying to obtain the collagen peptide-zinc chelate.
In the step 1, the processing environment temperature of the fresh shellfish mantle is 2-4 ℃.
Removing the hybrid protein in the step 2 specifically comprises the following steps: and (2) adding 0.08-0.12 mol/L NaOH solution into the raw materials obtained in the step (1), stirring for 24-48 h in an environment with the temperature of 2-4 ℃, centrifuging, removing supernatant, and repeatedly adding distilled water to clean the raw materials to enable the raw materials to be neutral, thus obtaining the collagen.
The mass ratio of the raw materials to the NaOH solution is 1: 8-10.
Step 2, preparing the denatured collagen by a heat extraction method, which specifically comprises the following steps: soaking the obtained collagen with distilled water, extracting at the water bath temperature of 90-100 ℃, filtering to remove insoluble substances, and freeze-drying to obtain the modified collagen.
The mass ratio of the collagen to the distilled water is 1: 8-10.
The step 3 specifically comprises the following steps: dissolving the denatured collagen in distilled water at a concentration of 5-15 mg/mL, adjusting the pH to 2-3 or 7.5-8.5, adding pepsin or trypsin accounting for 1-2% of the mass of the collagen respectively, placing the mixture in a shaking table for enzymolysis for 4-6 hours, then performing heat treatment to inactivate the enzyme, and performing freeze drying to obtain the collagen peptide after enzymolysis.
The step 4 specifically comprises the following steps: dissolving collagen peptide in distilled water, adjusting the pH value to 5.5-6.5, mixing the collagen peptide solution with a zinc salt solution, stirring for dissolving, chelating for 35-45 min under the condition of water bath oscillation at 45-60 ℃, then adding absolute ethyl alcohol into the reaction solution, standing for 45-60 min in an environment at 25-30 ℃, centrifugally separating precipitates, repeatedly cleaning the precipitates with the absolute ethyl alcohol, collecting the precipitates, and drying at 40-50 ℃ to obtain the collagen peptide-zinc chelate.
The concentration of the collagen peptide solution is 100-200 mg/mL, the concentration of the zinc salt solution is 80-120 mg/mL, the volume ratio of the zinc salt solution to the collagen peptide solution is 1: 5-6, and the mass concentration of ethanol in the solution is 75-85% after absolute ethanol is added.
The method has the advantages that the method takes the processing waste of marine organisms as the raw material, has simple and convenient acquisition way, low cost and high chelating rate, and avoids the possible risks of mad cow disease, foot and mouth disease and the like in the collagen of the terrestrial animals. Meanwhile, the peptide-zinc chelate chelated by the collagen peptide and the zinc ions has the advantages of small toxic and side effects, easy absorption and difficult residue, can exert the synergistic effect of the collagen peptide and the zinc ions, and can provide safe and effective functional food for consumers.
Drawings
FIG. 1 is a Fourier infrared spectrum of a peptide-zinc chelate complex and a collagen peptide prepared in example 1 of the present invention.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention provides a preparation method of a collagen peptide-zinc chelate, which comprises the steps of obtaining denatured water-soluble collagen by a heat extraction method, hydrolyzing the water-soluble collagen by using protease to prepare collagen peptide, and chelating with metal zinc ions to prepare the collagen peptide-zinc chelate. The method is implemented by the following steps:
step 1: treatment of raw materials
Selecting a fresh pearl shell mantle, cleaning the mantle with distilled water to remove impurities, and cutting the mantle into blocks of 2cm multiplied by 2cm at the temperature of 2-4 ℃ to obtain the raw material.
Step 2: removal of hetero-proteins (proteins other than collagen)
Adding 0.08-0.12 mol/L NaOH solution into the raw materials obtained in the step 1, wherein the mass ratio of the raw materials to the NaOH solution is 1: 8-10, stirring and processing for 24-48 h in an environment with the temperature of 2-4 ℃, centrifuging at the rotating speed of 8000-12000 rpm, removing supernatant, and then repeatedly adding distilled water to clean the raw materials for 5 times or more to enable the raw materials to be close to neutrality, so as to obtain the pearl oyster mantle collagen.
And step 3: preparation of collagen after thermal denaturation
Soaking the pearl oyster mantle collagen obtained in the step 2 in distilled water, wherein the mass ratio of the feed liquid is 1: 8-10. Extracting at 90-100 ℃ water bath temperature. And finally, filtering to remove insoluble substances, and freeze-drying to obtain the hot water-extracted collagen.
And 4, step 4: obtaining collagen peptide by enzymolysis of collagen with pepsin or trypsin
Dissolving the hot-water-extracted collagen obtained in the step 3 in distilled water at a concentration of 5-15 mg/mL, adjusting the pH to 2-3 or 7.5-8.5 with hydrochloric acid, and adding pepsin or trypsin which accounts for 1-2% of the mass of the collagen. And (4) placing the mixture in a shaking table for enzymolysis for 4-6 hours. And the heat treatment enzyme deactivation parameter is that enzyme deactivation is carried out for 10min at the temperature of 95 ℃, then the pearl oyster is naturally cooled to the room temperature and then frozen, and the enzymolysis pearl oyster mantle collagen peptide is obtained after freeze drying.
And 5: collagen peptide-zinc chelation reaction
And (4) dissolving the collagen peptide prepared in the step (4) in distilled water to prepare a collagen peptide solution with the collagen peptide concentration of 100-200 mg/mL, and adjusting the pH value to 5.5-6.5 by using hydrochloric acid. Dissolving zinc salt in distilled water to prepare a solution with the concentration of 80-120 mg/mL. Mixing the zinc salt solution and the collagen peptide solution according to the volume ratio of 1: 5-6, stirring and dissolving, and chelating for 35-45 min under the condition of water bath oscillation at 45-60 ℃ for 90-120 r/min.
Step 6: preparation of peptide-zinc chelate
And (3) adding absolute ethyl alcohol into the mixed solution obtained in the step (5) to enable the mass concentration of the ethyl alcohol in the solution to reach 75-85%, standing for 45-60 min in an environment with the temperature of 25-30 ℃, centrifugally separating precipitates at 8000-12000 rpm, repeatedly cleaning the precipitates with the absolute ethyl alcohol, and centrifuging for 5 times or more. Collecting the precipitate, and drying at 40-50 ℃ to obtain the peptide-zinc chelate.
The method takes the processing waste of marine organisms as the raw material, and has the advantages of simple and convenient acquisition way, low cost, high chelating rate and capability of avoiding the risks of mad cow disease, foot and mouth disease and the like possibly occurring in the collagen of the terrestrial animals. Meanwhile, the peptide-zinc chelate chelated by the collagen peptide and the zinc ions has the advantages of small toxic and side effects, easy absorption and difficult residue, and can exert the synergistic effect of the collagen peptide and the zinc ions, and the prepared V-shaped shellfish collagen peptide-zinc chelate has good effect on promoting the formation of bones. Can provide safe and effective functional food for consumers.
Example 1:
selecting 100g of fresh pearl shell mantle, cleaning the mantle with distilled water to remove impurities, and cutting the mantle into blocks of 2cm multiplied by 2cm at the temperature of 2-4 ℃ to obtain the raw material. Adding 0.1mol/L NaOH solution into the prepared pearl shell mantle, stirring and processing the raw material and the NaOH solution for 24 hours in an environment with the temperature of 4 ℃, centrifuging at the rotating speed of 10000rpm, removing supernatant, repeatedly adding distilled water to wash the raw material for 5 times to enable the raw material to be neutral, soaking the pretreated pearl shell mantle with the distilled water, performing water bath extraction in the environment with the temperature of 90 ℃, finally performing suction filtration to remove insoluble substances, and freeze-drying the filtrate to obtain the hot water-extracted collagen. Dissolving the prepared collagen in distilled water at a concentration of 10mg/mL, adjusting pH to 7.5, adding trypsin with a mass of 1.5% of that of the collagen, placing the mixture in a shaking table for enzymolysis for 5 hours, then inactivating the enzyme at 95 ℃ for 10min, and freeze-drying to obtain 1.82g of collagen peptide after enzymolysis.
Dissolving 500mg of prepared collagen peptide in 5mL of distilled water to prepare a collagen peptide solution, adjusting the pH value to 6, dissolving zinc sulfate in distilled water to prepare a solution with the concentration of 100mg/mL, stirring and mixing a salt solution and the collagen peptide solution according to the proportion of 1:5.5, carrying out chelation reaction for 45min under the condition of water bath oscillation at the temperature of 45 ℃, then adding absolute ethyl alcohol into the mixed solution to ensure that the final concentration of the ethyl alcohol in the solution reaches 80%, standing for 45min at the temperature of 30 ℃, centrifuging for 10min at 10000rpm to obtain a precipitate, then repeatedly cleaning the precipitate with the absolute ethyl alcohol, and centrifuging for 5 times. The precipitate was collected and dried at 50 ℃ to obtain a peptide-zinc chelate complex, and 0.122g of the chelate complex was finally obtained.
Example 2 (comparative example):
dissolving commercially available tilapia collagen peptide 500mg in 5mL of distilled water to prepare a collagen peptide solution, adjusting pH to 6, dissolving zinc sulfate in distilled water to prepare a solution with the concentration of 100mg/mL, mixing the zinc sulfate solution and the collagen peptide solution according to the volume ratio of 1:5.5, stirring for dissolving, carrying out chelation reaction for 45min at the temperature of 45 ℃, then adding absolute ethyl alcohol into the mixed solution to enable the final concentration of the ethyl alcohol in the solution to reach 80%, standing for 45min at the temperature of 30 ℃, centrifuging for 10min at 10000rpm to obtain a precipitate, then repeatedly cleaning the precipitate with the absolute ethyl alcohol, and centrifuging for 5 times. The precipitate was collected and dried at 50 ℃ to obtain a peptide-zinc chelate complex, and 0.093g of the chelate complex was finally obtained.
Example 3:
selecting 100g of fresh pearl shell mantle, cleaning the mantle with distilled water to remove impurities, and cutting the mantle into blocks of 2cm multiplied by 2cm at the temperature of 2-4 ℃ to obtain the raw material. Adding 0.08mol/L NaOH solution into the prepared pearl shell mantle, stirring and processing the pearl shell mantle and the NaOH solution at a mass ratio of 1:8 in an environment with the temperature of 2 ℃ for 48h, then centrifuging at a rotating speed of 12000rpm, removing supernatant, then repeatedly adding distilled water to wash the raw material for 7 times to make the raw material neutral, soaking the pretreated pearl shell mantle in the distilled water at a mass ratio of 1:8, carrying out water bath extraction in an environment with the temperature of 100 ℃, finally carrying out suction filtration to remove insoluble substances, and freeze-drying to obtain the collagen. Dissolving the prepared collagen in distilled water at a concentration of 15mg/mL, adjusting pH to 7.0, adding pepsin with a mass of 2% of the collagen, placing in a shaking table for enzymolysis for 6 hours, then inactivating enzyme at 90 ℃ for 10min, and freeze-drying to obtain 1.79g of collagen peptide after enzymolysis.
Dissolving 750mg of prepared pearl oyster mantle collagen peptide in 5mL of distilled water to prepare a collagen peptide solution, adjusting the pH to 6.5, dissolving zinc sulfate in the distilled water to prepare a solution with the concentration of 200mg/mL, mixing a salt solution and the collagen peptide solution according to the volume ratio of 1:6, stirring for dissolving, carrying out chelation reaction for 35min under the condition of water bath oscillation at the temperature of 60 ℃ for 120r/min, then adding absolute ethyl alcohol into the mixed solution to enable the final concentration of the ethyl alcohol in the solution to reach 85%, standing for 60min in the environment of 25 ℃, centrifuging for 10min at 12000rpm to obtain a precipitate, then repeatedly cleaning and precipitating by using the absolute ethyl alcohol, and centrifuging for 6 times. The precipitate was collected and dried at 40 ℃ to obtain a peptide-zinc chelate complex, and 0.191g of the chelate complex was finally obtained.
Example 4
Selecting 100g of fresh pearl shell mantle, cleaning the mantle with distilled water to remove impurities, and cutting the mantle into blocks of 2cm multiplied by 2cm at the temperature of 2-4 ℃ to obtain the raw material. Adding 0.12mol/L NaOH solution into the prepared pearl shell mantle, stirring and processing the raw material and the NaOH solution for 36 hours in an environment with the temperature of 3 ℃, centrifuging at the rotating speed of 8000rpm, removing supernatant, repeatedly adding distilled water to wash the raw material for 6 times to enable the raw material to be neutral, soaking the pretreated pearl shell mantle with the distilled water, performing water bath extraction in the environment with the temperature of 95 ℃, finally performing suction filtration to remove insoluble substances, and freeze-drying the filtrate to obtain the hot water-extracted collagen. Dissolving the prepared collagen in distilled water at a concentration of 5mg/mL, adjusting pH to 5, adding trypsin with a mass of 1% of that of the collagen, placing the mixture in a shaking table for enzymolysis for 4 hours, then inactivating the enzyme at 90 ℃ for 10min, and freeze-drying to obtain 1.32g of collagen peptide after enzymolysis.
Dissolving 1000mg of prepared collagen peptide in 5mL of distilled water to prepare a collagen peptide solution, adjusting the pH to 5.5, dissolving zinc sulfate in the distilled water to prepare a solution with the concentration of 80mg/mL, mixing a salt solution and the collagen peptide solution according to the volume ratio of 1:5, stirring and dissolving, carrying out chelation reaction for 40min under the condition of water bath oscillation at the temperature of 55 ℃ for 100r/min, then adding absolute ethyl alcohol into the mixed solution to ensure that the final concentration of the ethyl alcohol in the solution reaches 75%, standing for 50min at the temperature of 27 ℃, centrifuging for 10min at 8000rpm to obtain a precipitate, then repeatedly cleaning the precipitate with the absolute ethyl alcohol, and centrifuging for 7 times. The precipitate was collected and dried at 45 ℃ to obtain a peptide-zinc chelate complex, and 0.201g of the chelate complex was finally obtained.
It can be seen from the above examples that the collagen peptide prepared by the present invention has a higher zinc chelation rate than other examples.
The pearl oyster mantle collagen peptide prepared by the method of the invention has the following amino acid composition:
10mg of the collagen peptide prepared in example 1 was mixed with 2mL of a hydrochloric acid solution having a concentration of 6moL/L, and the mixture was hydrolyzed at 110 ℃ for 24 hours. The results of the amino acid analysis are shown in Table 1.
TABLE 1 amino acid composition of collagen peptides
As can be seen from the table, the most abundant amino acid among all amino acids is glycine, which accounts for about 30% of the total amino acid amount, and proline and hydroxyproline, which account for about 20% of the total amino acid amount, and have collagen-specific properties. In addition, compared with type I collagen, the collagen has higher contents of acidic amino acid, tryptophan and leucine, and is beneficial to chelating with zinc ions.
The peptide-zinc chelate complex and collagen peptide prepared in example 1 were analyzed for their patterns by Fourier Infrared Spectroscopy (FTIR), as shown in FIG. 1. Collagen peptide at 1655cm-1Has characteristic absorption peaks caused by carbonyl stretching vibration and amino bending vibration. After chelating zinc ions, the peak value is shifted to the lower 1639cm-1At frequency, these results indicate that the carbonyl groups in the peptide sequester zinc ions. In addition, two more significant vibration frequencies were also changed, with the peak of carboxyl groups from 1404cm in collagen peptide-1Moved to 1384cm-1And the peak value of the hydroxyl group is 3408cm-1(collagen peptide) shifted to a higher frequency of 3446cm-1(peptide-zinc chelate). These results indicate that zinc chelated collagen peptides are primarily chelated to zinc ions via the carbonyl, carboxyl, and hydroxyl oxygen in peptide bonds.
In order to further verify the bone formation promoting effect of the peptide-zinc chelate complex of the present invention, in vitro cell tests were performed, and the test methods and results were as follows:
1. MC3T3-E1 cell culture
The mouse skull-derived preosteoblasts MC3T3-E1 were cultured in α -MEM medium containing 10% FBS and 1% penicillin and streptomycin at 37 ℃ in a 5% carbon dioxide saturated humidity incubator. When the cell confluence reached 90%, the cells were digested with 0.25% pancreatin and then passaged at a ratio of 1: 3.
2. Test methods and results
(1) Dose setting and grouping: the cells were divided into 4 groups of 3-5 parallel groups, collagen peptide group (50. mu.g/mL), peptide-zinc chelate group (50. mu.g/mL), zinc ion group (1. mu.M/L), and blank control group.
(2) Osteoblast proliferation assay
Cell concentration was adjusted to 1.5X 10 with alpha-MEM medium4And (3) inoculating 100 mu L of the suspension into a 96-well plate, culturing for 24h, then discarding the stock culture solution, adding different complete growth culture media in groups according to the scheme, continuing to culture for 2d, then adding 20 mu L (5mg/mL) of MTT into each well, culturing for 4h, discarding the culture media, adding 150 mu L of DMSO, fully dissolving blue formazan crystals, and measuring the absorbance value at 490 nm.
Cell proliferation rate (%). sample group OD490nmValue/blank control OD490nmValue x 100
And (3) test results: the results of the effect of each group of drugs on cell proliferation are shown in Table 2.
TABLE 2 results of the Effect of the groups of drugs on cell proliferation
P <0.05, compared to placebo
The early experimental results show that different doses of zinc ions have different promoting effects on the proliferation of osteoblasts, wherein the effect is optimal under the condition of 1 mu M/L, and the promoting effect is reduced after the concentration is continuously increased, so that the zinc ions with the concentration of 1uM/L are selected for comparison. The collagen peptide treated cells did not significantly promote the proliferation of the cells compared to the blank control group, while both the peptide-zinc chelate group and the zinc ion group significantly promoted the proliferation of the cells (P < 0.05). The proliferation of the cells is obviously better than that of the zinc ion group by the peptide-zinc chelating group.
(3) Effect of osteoblast ALP Activity
Adjusting the cell density to 2X 104Each well was inoculated into a 24-well plate at 1mL per well, and after 24 hours, different differentiation media (containing 10mM sodium β -glycerophosphate and 50. mu.g/mL ascorbic acid) were added in groups according to the protocol described above, during which time the differentiation media containing the samples were exchanged at 2d intervals, and the media was discarded after 7d of incubation. After washing twice with a cooled PBS solution, 200. mu.L of a cell lysate (1% TritonX-100) was added to each well, lysed at 4 ℃ for 30min, and then centrifuged at 12000g/min for 5min (4 ℃), and the supernatant was taken, and the ALP content and the protein content were measured by the BCA kit. The activity of alkaline phosphatase is expressed as the activity of the enzyme released per gram of protein.
And (3) test results: the results of the effect of each group of drugs on cell proliferation are shown in Table 3.
TABLE 3 Effect of various groups of samples on promotion of ALP Activity of cells
P <0.05, compared to placebo
ALP is a marker product of osteoblast differentiation. Analyzing the results of table 3 reveals: compared with the blank control group, the collagen peptide group and the zinc ion group can not remarkably promote the differentiation of the cell ALP, but the peptide-zinc chelate group can remarkably promote the activity of the cell ALP.
Claims (10)
1. A preparation method of a collagen peptide-zinc chelate is characterized in that fresh shellfish is used as a raw material, a heat extraction method is adopted to obtain water-soluble collagen of a shellfish mantle after denaturation, then protease is used for hydrolyzing the water-soluble collagen to prepare collagen peptide, and the collagen peptide is chelated with metal zinc ions to prepare the collagen peptide-zinc chelate.
2. The method for preparing a collagen peptide-zinc chelate according to claim 1, which is carried out by the following steps:
step 1, cleaning a fresh shellfish mantle with distilled water, removing impurities, and processing into small raw materials;
step 2, removing foreign proteins in the raw materials processed in the step 1 to obtain collagen, and preparing denatured collagen by adopting a thermal extraction method;
step 3, carrying out enzymolysis on the denatured collagen obtained in the step 2 by adopting pepsin or trypsin to obtain collagen peptide;
and 4, carrying out chelation reaction on the collagen peptide obtained in the step 3 and a zinc salt solution, separating and drying to obtain the collagen peptide-zinc chelate.
3. The method for preparing a collagen peptide-zinc chelate according to claim 2, wherein the processing environment temperature of the fresh shellfish mantle in the step 1 is 2-4 ℃.
4. The method for preparing a collagen peptide-zinc chelate according to claim 2, wherein the removal of the hetero-proteins in the step 2 specifically comprises: and (2) adding 0.08-0.12 mol/L NaOH solution into the raw materials obtained in the step (1), stirring for 24-48 h in an environment with the temperature of 2-4 ℃, centrifuging, removing supernatant, and repeatedly adding distilled water to clean the raw materials to enable the raw materials to be neutral, thus obtaining the collagen.
5. The method for preparing the collagen peptide-zinc chelate complex according to claim 4, wherein the mass ratio of the raw material to the NaOH solution is 1: 8-10.
6. The method for preparing a collagen peptide-zinc chelate according to claim 2, wherein the heat extraction method in step 2 is used for preparing denatured collagen, and specifically comprises the following steps: soaking the obtained collagen with distilled water, extracting at the water bath temperature of 90-100 ℃, filtering to remove insoluble substances, and freeze-drying to obtain the modified collagen.
7. The method for preparing the collagen peptide-zinc chelate according to claim 6, wherein the mass ratio of the collagen to the distilled water is 1: 8-10.
8. The method for preparing a collagen peptide-zinc chelate according to claim 2, wherein the step 3 specifically comprises: dissolving the denatured collagen in distilled water at a concentration of 5-15 mg/mL, adjusting the pH to 2-3 or 7.5-8.5, adding pepsin or trypsin accounting for 1-2% of the mass of the collagen respectively, placing the mixture in a shaking table for enzymolysis for 4-6 hours, then performing heat treatment to inactivate the enzyme, and performing freeze drying to obtain the collagen peptide after enzymolysis.
9. The method for preparing a collagen peptide-zinc chelate according to claim 2, wherein the step 4 specifically comprises: dissolving collagen peptide in distilled water, adjusting the pH value to 5.5-6.5, mixing the collagen peptide solution with a zinc salt solution, stirring for dissolving, chelating for 35-45 min under the condition of water bath oscillation at 45-60 ℃, then adding absolute ethyl alcohol into the reaction solution, standing for 45-60 min in an environment at 25-30 ℃, centrifugally separating precipitates, repeatedly cleaning the precipitates with the absolute ethyl alcohol, collecting the precipitates, and drying at 40-50 ℃ to obtain the collagen peptide-zinc chelate.
10. The method for preparing a collagen peptide-zinc chelate according to claim 9, wherein the concentration of the collagen peptide solution is 100 to 200mg/mL, the concentration of the zinc salt solution is 80 to 120mg/mL, the volume ratio of the zinc salt solution to the collagen peptide solution is 1:5 to 6, and the mass concentration of ethanol in the solution after adding absolute ethanol is 75 to 85%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910783971.0A CN110590938A (en) | 2019-08-23 | 2019-08-23 | Preparation method of collagen peptide-zinc chelate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910783971.0A CN110590938A (en) | 2019-08-23 | 2019-08-23 | Preparation method of collagen peptide-zinc chelate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110590938A true CN110590938A (en) | 2019-12-20 |
Family
ID=68855328
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910783971.0A Pending CN110590938A (en) | 2019-08-23 | 2019-08-23 | Preparation method of collagen peptide-zinc chelate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110590938A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111374307A (en) * | 2020-03-30 | 2020-07-07 | 江西师范大学 | Anti-digestion fish skin gelatin composite gel zinc-supplementing nutrition bag and preparation method and application thereof |
| CN111387508A (en) * | 2020-03-12 | 2020-07-10 | 大连工业大学 | Zinc chelating peptide gel and preparation method thereof |
| CN114703245A (en) * | 2022-04-14 | 2022-07-05 | 甘肃中肽生物科技有限公司 | Preparation process of bovine bone protein peptide chelated zinc |
| CN116076738A (en) * | 2022-11-30 | 2023-05-09 | 大连工业大学 | Water-in-oil-in-water double-layer emulsion embedded with peptide zinc chelate and preparation method thereof |
| CN116970072A (en) * | 2023-09-22 | 2023-10-31 | 烟台源力德海洋生物有限公司 | Preparation method of collagen peptide |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004049046A (en) * | 2002-07-17 | 2004-02-19 | Nippon Zenyaku Kogyo Kk | Solid formulation for livestock |
| CN101457249A (en) * | 2009-01-05 | 2009-06-17 | 海南大学 | Method for preparing indissoluble aquatic collagen protein source ACE inhibitor peptides |
| CN103626867A (en) * | 2013-06-19 | 2014-03-12 | 中国海洋大学 | Preparation process for zinc fish-skin collagen polypeptide chelate |
-
2019
- 2019-08-23 CN CN201910783971.0A patent/CN110590938A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004049046A (en) * | 2002-07-17 | 2004-02-19 | Nippon Zenyaku Kogyo Kk | Solid formulation for livestock |
| CN101457249A (en) * | 2009-01-05 | 2009-06-17 | 海南大学 | Method for preparing indissoluble aquatic collagen protein source ACE inhibitor peptides |
| CN103626867A (en) * | 2013-06-19 | 2014-03-12 | 中国海洋大学 | Preparation process for zinc fish-skin collagen polypeptide chelate |
Non-Patent Citations (1)
| Title |
|---|
| 马婷，等: "珍珠贝外套膜胶原蛋白肽及其锌螯合物的体外抑制骨质疏松作用", 《肉类研究》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111387508A (en) * | 2020-03-12 | 2020-07-10 | 大连工业大学 | Zinc chelating peptide gel and preparation method thereof |
| CN111374307A (en) * | 2020-03-30 | 2020-07-07 | 江西师范大学 | Anti-digestion fish skin gelatin composite gel zinc-supplementing nutrition bag and preparation method and application thereof |
| CN114703245A (en) * | 2022-04-14 | 2022-07-05 | 甘肃中肽生物科技有限公司 | Preparation process of bovine bone protein peptide chelated zinc |
| CN116076738A (en) * | 2022-11-30 | 2023-05-09 | 大连工业大学 | Water-in-oil-in-water double-layer emulsion embedded with peptide zinc chelate and preparation method thereof |
| CN116970072A (en) * | 2023-09-22 | 2023-10-31 | 烟台源力德海洋生物有限公司 | Preparation method of collagen peptide |
| CN116970072B (en) * | 2023-09-22 | 2023-12-15 | 烟台源力德海洋生物有限公司 | Preparation method of collagen peptide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110590938A (en) | Preparation method of collagen peptide-zinc chelate | |
| WO2019119998A1 (en) | Protein-derived, heavy metal removal agent suitable for use in mussels and preparation method therefor | |
| CN103386144B (en) | Preparation method of fish collagen combined chitosan biological dressing | |
| CN101904491B (en) | Process for producing cattle or sheep chondroitin sulfate | |
| CN101851300A (en) | Process for extracting chondroitin sulfate | |
| CN112795612B (en) | Preparation method for extracting peptide chelated calcium by combined conversion of enzymolysis and chelation | |
| CN1771930A (en) | Externally applied compound amino acid prepn for promoting wound healing and its prepn and application | |
| CN102134288B (en) | Process for extracting chondroitin sulfate from pigs | |
| WO2023040083A1 (en) | Method for preparing sea bream collagen peptide | |
| CN109680027B (en) | Grifola frondosa small peptide iron chelate as well as preparation method and application thereof | |
| CN107674905A (en) | Spirulina bioactive peptide, composition and preparation method | |
| CN107619411B (en) | Heme extraction method | |
| CN109258916A (en) | A kind of fish scale collagen peptide calcium and preparation method thereof, purposes | |
| CN113151385A (en) | Method for efficiently preparing livestock and poultry cartilage collagen polypeptide | |
| CN112956702A (en) | Compound preparation for increasing bone mineral density | |
| CN115669839B (en) | Bovine bone collagen peptide-calcium chelate solid beverage and preparation method thereof | |
| CN111635919A (en) | Method for preparing collagen oligopeptide by hydrolyzing animal skin with bacillus subtilis | |
| CN1814782A (en) | Method for extracting small molecule collagen protein from fish scale utilizing enzyme engineering technique | |
| CN113350381B (en) | Method for increasing protein ratio of pearl hydrolysate and treatment solution | |
| CN115449535A (en) | Bone-derived collagen tripeptide and preparation method thereof | |
| CN110527705B (en) | Method for preparing antioxidant oligopeptide by enzymolysis of rabbit blood fermentation broth | |
| CN113444143B (en) | Camel bone antioxidant polypeptide and preparation method and application thereof | |
| WO2023082523A1 (en) | Method for improving extraction rate of chondroitin sulfate prepared from tilapia skull | |
| CN112941134A (en) | Whole sheep small molecule peptide extraction process | |
| CN112245452A (en) | Preparation method of bone extract and application of bone extract to osteoporosis and osteoarthritis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191220 |