CN108743446A - A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application - Google Patents
A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application Download PDFInfo
- Publication number
- CN108743446A CN108743446A CN201810686464.0A CN201810686464A CN108743446A CN 108743446 A CN108743446 A CN 108743446A CN 201810686464 A CN201810686464 A CN 201810686464A CN 108743446 A CN108743446 A CN 108743446A
- Authority
- CN
- China
- Prior art keywords
- protein
- silk
- cocoon
- obtains
- ultrafiltration membrane
- 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
- 238000000108 ultra-filtration Methods 0.000 title claims abstract description 29
- 108010013296 Sericins Proteins 0.000 title claims abstract description 27
- 239000012528 membrane Substances 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 77
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 77
- 239000003292 glue Substances 0.000 claims abstract description 63
- 238000000034 method Methods 0.000 claims abstract description 45
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims abstract description 41
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000006228 supernatant Substances 0.000 claims abstract description 22
- 229910000029 sodium carbonate Inorganic materials 0.000 claims abstract description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000000284 extract Substances 0.000 claims abstract description 16
- 238000005238 degreasing Methods 0.000 claims abstract description 14
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 238000005119 centrifugation Methods 0.000 claims abstract description 13
- 239000002537 cosmetic Substances 0.000 claims abstract description 11
- 235000013305 food Nutrition 0.000 claims abstract description 11
- 239000003208 petroleum Substances 0.000 claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 9
- 238000004108 freeze drying Methods 0.000 claims abstract description 9
- 239000000287 crude extract Substances 0.000 claims description 28
- 239000004615 ingredient Substances 0.000 claims description 12
- 238000000751 protein extraction Methods 0.000 claims description 11
- 150000003384 small molecules Chemical class 0.000 claims description 11
- 239000012535 impurity Substances 0.000 claims description 10
- 239000000049 pigment Substances 0.000 claims description 10
- 238000002203 pretreatment Methods 0.000 claims description 10
- 238000010992 reflux Methods 0.000 claims description 10
- 238000007873 sieving Methods 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 238000004806 packaging method and process Methods 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 4
- 230000001857 anti-mycotic effect Effects 0.000 claims description 2
- 239000002543 antimycotic Substances 0.000 claims description 2
- 235000013373 food additive Nutrition 0.000 claims description 2
- 239000002778 food additive Substances 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims 1
- 108010022355 Fibroins Proteins 0.000 abstract description 72
- 239000000047 product Substances 0.000 abstract description 15
- -1 centrifugation Substances 0.000 abstract description 10
- 238000010612 desalination reaction Methods 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000009826 distribution Methods 0.000 abstract description 4
- 239000002699 waste material Substances 0.000 abstract description 4
- 239000012141 concentrate Substances 0.000 abstract description 2
- 239000000706 filtrate Substances 0.000 abstract description 2
- 238000001694 spray drying Methods 0.000 abstract description 2
- 238000007670 refining Methods 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 69
- 239000000243 solution Substances 0.000 description 64
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 37
- 229910001868 water Inorganic materials 0.000 description 35
- 239000000523 sample Substances 0.000 description 33
- 239000000843 powder Substances 0.000 description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 108090000765 processed proteins & peptides Proteins 0.000 description 17
- 235000001014 amino acid Nutrition 0.000 description 16
- 150000001413 amino acids Chemical class 0.000 description 15
- 230000007062 hydrolysis Effects 0.000 description 14
- 238000006460 hydrolysis reaction Methods 0.000 description 14
- 239000000463 material Substances 0.000 description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 239000007788 liquid Substances 0.000 description 11
- 229910052757 nitrogen Inorganic materials 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 239000000499 gel Substances 0.000 description 8
- 239000000017 hydrogel Substances 0.000 description 8
- 238000000926 separation method Methods 0.000 description 8
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 7
- 230000008901 benefit Effects 0.000 description 7
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 7
- 229910052739 hydrogen Inorganic materials 0.000 description 7
- 239000003595 mist Substances 0.000 description 7
- QKFJKGMPGYROCL-UHFFFAOYSA-N phenyl isothiocyanate Chemical compound S=C=NC1=CC=CC=C1 QKFJKGMPGYROCL-UHFFFAOYSA-N 0.000 description 7
- 239000007921 spray Substances 0.000 description 7
- LOTKRQAVGJMPNV-UHFFFAOYSA-N 1-fluoro-2,4-dinitrobenzene Chemical compound [O-][N+](=O)C1=CC=C(F)C([N+]([O-])=O)=C1 LOTKRQAVGJMPNV-UHFFFAOYSA-N 0.000 description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 6
- RAHZWNYVWXNFOC-UHFFFAOYSA-N Sulphur dioxide Chemical compound O=S=O RAHZWNYVWXNFOC-UHFFFAOYSA-N 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 210000003491 skin Anatomy 0.000 description 6
- 239000012224 working solution Substances 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 235000011167 hydrochloric acid Nutrition 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 230000003647 oxidation Effects 0.000 description 5
- 238000007254 oxidation reaction Methods 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 208000002109 Argyria Diseases 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 4
- 241000270708 Testudinidae Species 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 210000004209 hair Anatomy 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 150000003254 radicals Chemical class 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 241000270607 Chelonia mydas Species 0.000 description 3
- 102000008186 Collagen Human genes 0.000 description 3
- 108010035532 Collagen Proteins 0.000 description 3
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 3
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 3
- 239000005864 Sulphur Substances 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 229920001436 collagen Polymers 0.000 description 3
- 229960003067 cystine Drugs 0.000 description 3
- 238000001212 derivatisation Methods 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 238000001641 gel filtration chromatography Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 235000015110 jellies Nutrition 0.000 description 3
- 239000008274 jelly Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 229910052717 sulfur Inorganic materials 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 108010039627 Aprotinin Proteins 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 238000007445 Chromatographic isolation Methods 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 102000016943 Muramidase Human genes 0.000 description 2
- 108010014251 Muramidase Proteins 0.000 description 2
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 description 2
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 2
- 108010058846 Ovalbumin Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical class [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101710162629 Trypsin inhibitor Proteins 0.000 description 2
- 229940122618 Trypsin inhibitor Drugs 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002745 absorbent Effects 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 description 2
- 229960004405 aprotinin Drugs 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- AFYNADDZULBEJA-UHFFFAOYSA-N bicinchoninic acid Chemical compound C1=CC=CC2=NC(C=3C=C(C4=CC=CC=C4N=3)C(=O)O)=CC(C(O)=O)=C21 AFYNADDZULBEJA-UHFFFAOYSA-N 0.000 description 2
- 239000012159 carrier gas Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 238000005336 cracking Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 235000019621 digestibility Nutrition 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 239000010419 fine particle Substances 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 239000004325 lysozyme Substances 0.000 description 2
- 229960000274 lysozyme Drugs 0.000 description 2
- 235000010335 lysozyme Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 229940092253 ovalbumin Drugs 0.000 description 2
- 229940117953 phenylisothiocyanate Drugs 0.000 description 2
- 239000003531 protein hydrolysate Substances 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000012723 sample buffer Substances 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002753 trypsin inhibitor Substances 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- HVBSAKJJOYLTQU-UHFFFAOYSA-N 4-aminobenzenesulfonic acid Chemical compound NC1=CC=C(S(O)(=O)=O)C=C1 HVBSAKJJOYLTQU-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 208000003556 Dry Eye Syndromes Diseases 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical class O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000004195 Isomerases Human genes 0.000 description 1
- 108090000769 Isomerases Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 239000011837 N,N-methylenebisacrylamide Substances 0.000 description 1
- FZERHIULMFGESH-UHFFFAOYSA-N N-phenylacetamide Chemical compound CC(=O)NC1=CC=CC=C1 FZERHIULMFGESH-UHFFFAOYSA-N 0.000 description 1
- 229910004835 Na2B4O7 Inorganic materials 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 102000005158 Subtilisins Human genes 0.000 description 1
- 108010056079 Subtilisins Proteins 0.000 description 1
- 229920004933 Terylene® Polymers 0.000 description 1
- 102000005924 Triose-Phosphate Isomerase Human genes 0.000 description 1
- 108700015934 Triose-phosphate isomerases Proteins 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- WJEIYVAPNMUNIU-UHFFFAOYSA-N [Na].OC(O)=O Chemical compound [Na].OC(O)=O WJEIYVAPNMUNIU-UHFFFAOYSA-N 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 229910001870 ammonium persulfate Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 229910052786 argon Inorganic materials 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 238000003339 best practice Methods 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- VQLYBLABXAHUDN-UHFFFAOYSA-N bis(4-fluorophenyl)-methyl-(1,2,4-triazol-1-ylmethyl)silane;methyl n-(1h-benzimidazol-2-yl)carbamate Chemical compound C1=CC=C2NC(NC(=O)OC)=NC2=C1.C=1C=C(F)C=CC=1[Si](C=1C=CC(F)=CC=1)(C)CN1C=NC=N1 VQLYBLABXAHUDN-UHFFFAOYSA-N 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 235000012970 cakes Nutrition 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015111 chews Nutrition 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000002316 cosmetic surgery Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- MOTZDAYCYVMXPC-UHFFFAOYSA-N dodecyl hydrogen sulfate Chemical compound CCCCCCCCCCCCOS(O)(=O)=O MOTZDAYCYVMXPC-UHFFFAOYSA-N 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 229910000743 fusible alloy Inorganic materials 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002923 metal particle Substances 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 1
- QLWXZRVOHCYKKK-UHFFFAOYSA-N n-(cyclohexylideneamino)-2,4-dinitroaniline Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC=C1NN=C1CCCCC1 QLWXZRVOHCYKKK-UHFFFAOYSA-N 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000002807 parathyroid gland hormone Substances 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 150000004968 peroxymonosulfuric acids Chemical class 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- CWHFDTWZHFRTAB-UHFFFAOYSA-N phenyl cyanate Chemical compound N#COC1=CC=CC=C1 CWHFDTWZHFRTAB-UHFFFAOYSA-N 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- HKOOXMFOFWEVGF-UHFFFAOYSA-N phenylhydrazine Chemical compound NNC1=CC=CC=C1 HKOOXMFOFWEVGF-UHFFFAOYSA-N 0.000 description 1
- 229940067157 phenylhydrazine Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000005020 polyethylene terephthalate Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical class [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000011091 sodium acetates Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 239000012209 synthetic fiber Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000010415 tropism Effects 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 210000004509 vascular smooth muscle cell Anatomy 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dermatology (AREA)
- Diabetes (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Food Science & Technology (AREA)
- Oncology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Communicable Diseases (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Zoology (AREA)
- Immunology (AREA)
- Nutrition Science (AREA)
- Birds (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of ultrafiltration membrane preparation method of cocoon sericin protein and its applications, this method will strip silk cocoon surface filiform, remove remaining magazine in silk cocoon, it is spare to be cut into strip, petroleum ether degreasing is added, soaked in absolute ethyl alcohol is decolourized, it recycles aqueous sodium carbonate to boil, extracts, ultrafiltration, merging filtrate, centrifugation, supernatant with use molecular cut off for the ultrafiltration membrane of 30KDa, progress desalination, it is detained solution and continues to concentrate, and obtains refining high-quality sericine using spray drying or freeze-drying;It is shown by SDS-PAAG results:The sericine of extraction contains protein of the molecular weight distribution within 14.4KDa-66KDa.Have the characteristics that glue protein matter yield is high, while ultrafiltration membrane reaches a step desalination and concentration glue protein matter purpose, the cocoon sericin albumen that this method obtains can be used in the raw material of cosmetics, food and medicine;Removal fibroin makes silk gloss in silk, and the silk gum after removal, which is waste, becomes a kind of high value addition product.
Description
Technical field
The present invention relates to a kind of ultrafiltration membrane preparation method of cocoon sericin protein and its applications.
Background technology
China's silk oilfields are abundant, are the big countries of silk production and processing, fibroin is that mankind's early start uses
One of native protein enjoys always the good reputation of " fiber queen " from ancient times to the present.The silk yield in China is in rank first,
Account for about the half of Gross World Product.It will produce a large amount of leftover bits and pieces, the profit again of silk offcut in the process of manufacture of silk
With being important research direction.Fibroin albumen is the important component of silk, is a kind of natural protein polymer macromolecular,
Submissive with strand, good biocompatibility is easy film forming, and machinability is good, and the features such as be easy to molecular modification, in biology
There is important application prospect in the fields such as material, medicine, environmental treatment, nano material, functional material.The extraction of fibroin albumen and silk
The research of plain biomembrane material has important theory significance and potential application value.Fibroin from silk is as a kind of
Novel protein resource, not only with edible value but also with medical value.
Protein content in silk is up to 97.6%-98.5%, and structure is mainly by two fibroins (Fibroin) and turnover
Silk gum (Sericin) two parts of covering form, and silk peptide belongs to the intermediate product of Fibroin Hydrolysis.Fibroin (Fibroin, fibroin
Albumen), the also known as silk heart is the chief component of silk, accounts about 70% one the 80% of silk content, is with crystalline texture
Protein, be made of 18 kinds of amino acid, wherein 8 kinds of essential amino acids necessary to human body account for 7% or so.Except C, H, O, N tetra-
Outside kind essential element, fibroin albumen also contains other elements such as Ca, Si, Sr, P etc., these elements determine the one of fibroin
A little performances.The amino acid composed structure of fibroin is relatively simple, and packing is easy between the peptide molecule formed, leads to have certain crystallization
Property.Crystallinity is in 50%-60% or so.Polarity side group accounts for about 29.5% in fibroin, and nonpolar side group accounts for about 70.5%, the two ratio
It is 0.42.And ratio between two is far above fibroin in silk gum, and about 2.91, this is also that silk gum is soluble easily in water and fibroin is not soluble in water simultaneously
Basic reason with some strength.Side group in fibroin is simple glycine, serine, alanine, by certain sequence
Structural arrangement is predominantly located at the crystal region of fibroin albumen at the segment of rule.And phenylalanine, color ammonia with larger side group
Acid, tyrosine etc. are then primarily present in amorphous regions.The conformation of fibroin albumen mainly based on a random ball of string, in addition also contains
There are a small amount of β-corner, alpha-helix etc., molecular weight to generally believe between the ranges of 3.6-3.7 × 10.5.
Silk is used as textile material in the past, gradually increases the research of silk with people in recent years, both at home and abroad to silkworm
The exploitation and utilization of silk and its product extend to the multiple fields such as food, material, drug, biomaterial and cosmetics.
1. the application of silk:
1.1 application in food:
Natural fibroin is a kind of azelon of macromolecular backbone, and quality is close, not soluble in water, is not only difficult to nozzle
It chews, human body stomach also can not directly absorb, only after fibroin molecule is degraded to lower molecular weight level by certain way,
Edible and the absorbable optimum efficiency utilized can be reached.Fibroin albumen through enzyme degrade or sour water solution after, can become oligopeptide with
The mixing of amino acid.Someone carried out with big white mouse animal experiments show that, big white mouse to enzyme degrade after silk fibroin powder digestion
Rate reaches 70%, reaches 91% to the digestibility of the silk fibroin powder after sour water solution;But if with silk fibroin water solution and not
The silk fibroin powder of degradation feeds big white mouse, and digestibility is respectively 47.4% and 27.6%.It can be seen that fibroin albumen
The fibroin albumen peptide and amino acid obtained after hydrolysis is easy to be digested absorption in vivo, this makes fibroin in functional food
Application aspect have larger potentiality to be exploited.Japan begins to set about the research of silk edibleization early in nineteen eighty-three, earliest
Product is that silk jelly has had developed a series of products using fibroin albumen by development for many years to current Japan.?
In the end of the year 1996, Japanese Jia Yue development companies come into effect the large-scale production of silk fibroin powder, and are sold to food producer.
There are many food of the kind containing fibroin powder such as gruel, beverage, fibroin albumen cake, noodles, candy, jelly pectin, soy sauce, ice rivers in Henan Province for oneself
The classes products such as leaching emerge, these products containing fibroin powder have become the health food of Japanese a new generation, and due to its unique product
Matter and it is favored by consumers.There is abundant waste silk resource, the research and development in relation to silk food also to have been reported in China.Lee
Moral is remote etc. will to pass through Na2CO3The fibroin protein obtained after solution degumming process is in CaCl2Further dissolution process in solution, warp
Silk protein solution is made in dialysis desalting.And beverage and jelly, good mouthfeel are prepared as raw material, transparent color and luster is eaten
It is safe and healthy.
1.2 application in the material:
In the traditional idea of people, silk is due to excellent wearability and environment amenable performance, making
The concern of people is constantly subjected to for a kind of natural fiber garment fabric.The hydrophilic-silk on silk surface is removed by degumming tech
After glue protein, the fibroin albumen of purifying can be obtained.Fibroin albumen has extremely excellent mechanical property:Tensile strength is reachable
To 0.5GPa, fracture elongation 15%.Excellent mechanical property possessed by fibroin albumen is inseparable with its own structure
's.Before nineteen ninety, Japan every year need to from national about 7.5 tons of the import green turtle crust such as Cuba and Indonesia, but due to
Green turtle is endangered wild animal type, thus since《Washington treaty》After forbidding green turtle to merchandise at the international level,
Japanese tortoise plastron association entrusts with Ministry of Commerce and Industry suffered by four countries of the counties Xiang Chuan industrial research, proceeds by and is made using fibroin
The experimental study of tortoise plastron substitute.On October 30th, 1992 in Asahi Shimbun, announces that research succeeds.The way of the method is
By transparent fibroin film well prepared in advance, suction press is carried out in water vapour, then obtains plate similar with tortoise plastron
Later when material adsorbing metal particle or after coloured with dyestuff the distinctive color of tortoise plastron can be presented, made of this method in material
The suture that craftwork enters behind market used in surgical operation welcomed by consumers generally is Silk Threads, although nearest Buddhist nun
The use of the synthetic fibers such as dragon, terylene is being continuously increased, but due to some superperformances such as fineness that silk suture has is small,
Easily knotting, tieing are not easy to scatter, and tensile strength is greatly, to human body without anaphylaxis etc. so that silk suture is in terms of some than closing
Advantage is had more at fiber, so the use of Silk Threads still occupies sizable ratio, cannot especially there is the ophthalmology of scar in wound
And plastic surgery, silk suture small using fineness, that tropism is good, intensity is big are more suitable.
1.3 application in medicine:
Fibroin albumen is also paid close attention to by more and more researchers in the application of biomedicine field, can be used as artificial skin
Skin, enzyme immobilizatio carrier, slow releasing carrier of medication and tissue engineering material etc..The hardness of cell culture medium be one in response to
The important physical factor of cell type, although the hydrogel prepared based on protein has been widely used as the culture of cell
Base, but their rigidity should further increase the Absorption Growth that can be just more conducive to cell.Lv et al. is with 1- (3- dimethylaminos
Propyl) for -3- ethyl-carbodiimide hydrochlorides by fibroin albumen and collagen induction crosslinking, preparation has the fibroin of appropriate hardness
Albumen/collagen mixing hydrogel.The storage modulus of the cross-linked hydrogel is higher than 3kPa, and even if above 10KPa, it still has very
High mechanical strength;In addition, cross-linked hydrogel remains to keep the conformation of its script not change at 80 DEG C, thus proving should
The heat-resistant stability of hydrogel is improved.Growth of the vascular smooth muscle cells in fibroin albumen/collagen hydrogels
Show that cross-linking reaction does not generate negative influence to the bioactivity of the hydrogel.The hydrogel will be in tissue engineering material
On have very wide application prospect.Its hydrolysate of fibroin albumen also has unique physiological activity and medical value, such as drops blood
Sugared effect, antioxidation, improves intestinal physiology function, the effect for preventing brain aging and cryoprotection work at norcholesterol effect
With etc..Therefore to fibroin albumen, this natural resources develops and utilizes, and not only improves warp of the fibroin as industry byproduct
Ji value, and there is larger Social benefit and economic benefit.
1.4 cosmetic field:
Fibroin albumen has good heat and moisture preserving, contributes to the moisture for adjusting skin, thus fibroin albumen can be used as
The good base material fibroin albumens of cosmetics are there are two types of the main application forms of cosmetic field:Fibroin powder and silk peptide.Naturally
Fibroin itself is not soluble in water, and fibroin powder remains the original structure composition of fibroin albumen, and therefore, fibroin powder inherits fibroin egg
While white distinctive sub-dued lustre, and the effect that ultraviolet light resists solar radiation that absorbs is had both.It is main in cosmetic field
Application be hydrolysis after relative molecular mass in 2KDa or less silk peptides.Wherein, the lower silk peptide of relative molecular mass
(300-800) can provide nutrient for the metabolism of skin and hair, and skin is contributed to resist the damage of chemistry and machinery with hair;Phase
It can then assign skin and hair natural gloss the silk peptide that molecular mass is 1000-2000Da, and then skin and hair is played
The effect of moisturizing.
2. hydrolysis and the separating-purifying of fibroin albumen:
The most crystal region of fibroin albumen macromolecular can not be utilized when not hydrolyzing, when it is hydrolyzed to molecular weight
When small molecule silk peptide less than 10KDa or so, the value of fibroin albumen could fully show.It, should be with fibroin in practical application
Based on the space structure of protein macromolecule main chain, using certain method for hydrolysis, fibroin albumen is hydrolyzed to average molecular matter
The smaller fibroin peptide product of amount, the silk peptide after hydrolysis is before biomedical, food and cosmetic field have wide application
Scape.The main method for hydrolysis of fibroin albumen has:Sour water solution, basic hydrolysis, enzyme hydrolysis etc..Each method has its advantage and disadvantage.Sour water
Solution can adequately be hydrolyzed fibroin albumen, but can lead to the destruction of amino acid;Basic hydrolysis is then easy to cause amino acid
Racemization, and then reduce fibroin albumen use value;Enzyme hydrolysis is due to having mild reaction condition and to Amino Acids Induced
Small advantage is expected to the best practice as hydrolytic degradation fibroin albumen.After fibroin albumen hydrolysis, should further it use corresponding
Separating-purifying means, the product after hydrolysis is purified.Currently, the method for protein hydrolysate separating-purifying is mainly wrapped
Include Flavonoids by Macroporous Adsorption Resin, gel filtration chromatography, high performance liquid chromatography and ultrafiltration etc..Macroporous absorbent resin itself has
Have a performance of absorption and screening, regenerating easily, have in purifies and separates bioactive substance simple equipment, mild condition and
Easy to operate advantage.Wu Jinhong, et al. using DA201-C macroporous absorbent resins to the sericin peptide taken of alcohol grading carry out desalination and
Separating-purifying, finally obtained polarity is different and has the sericin peptide taken of antioxidant activity, is the work(for being worth further investigation and exploitation
Energy product gel filtration chromatography has the advantages that simple, mild condition, and it is pure to carry out to be mainly based upon the molecular size range of sample
Change separation.Ni Li etc. isolates and purifies out silk peptide using gel filtration chromatography Sephadex-G15, finds alkali protease
When the degree of hydrolysis that Alcalase hydrolyzes fibroin is 20%, the ACE inhibitory activity of purifying gained silk peptide is most strong.Zhang Linghua is waited
Soya-bean polypeptides are isolated and purified using G-15 sephadexes.The application of high performance liquid chromatography (HPLC) not only can be with
It is completed in a short time separation purpose, it is often more important that HPLC can produce biologically active polypeptide on a preparative scale.It is super
Filter method is a kind of membrane separating method, is mainly used in genetic engineering field, is usually applied to desalination concentration, protein divides step by step
From and the processes such as buffer exchange.
Invention content
Present invention aims at provide ultrafiltration membrane preparation method and its application of a kind of cocoon sericin protein, this method
It will strip silk cocoon surface filiform, and remove remaining magazine in silk cocoon, it is spare to be cut into tiny strip, and oil is added at normal temperatures
Ether degreasing is decolourized after degreasing using soaked in absolute ethyl alcohol, and the sericine of the silk cocoon of degreasing, decoloration after dry is utilized carbonic acid
Sodium water solution boils, extraction, ultrafiltration, merging filtrate, centrifugation, supernatant with use molecular cut off for the ultrafiltration again of 30KDa
Film, rotating speed 3000rmp/min collect top, obtain glue protein extraction solution;It is shown through SDS-PAAG results:This method is extracted
Sericine contain protein of the molecular weight distribution within 14.4KDa-66KDa.The method of the invention has glue egg
The high feature of white matter yield, while ultrafiltration membrane reaches a step desalination and concentration glue protein matter purpose;Be a green, high yield,
Efficiently and it is easy to amplify a kind of high efficiency preparation method for the cocoon sericin protein implemented;The glue protein that this method obtains can be with
Used in cosmetics, the raw material of food and medicine;Removal fibroin makes silk gloss in silk, and the silk gum after removal, which is waste, becomes a kind of
High value addition product.
A kind of ultrafiltration membrane preparation method of cocoon sericin protein of the present invention, follows these steps to carry out:
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, the silk cocoon 100g that weighs that treated, with being dried after petroleum ether degreasing 48h, then again with soaked in absolute ethyl alcohol for 24 hours,
Small molecule ingredient and pigment are removed, is dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, the sodium carbonate liquor refluxing extraction 2-4 of a concentration of 0.05-0.1%
It is secondary, it 20 minutes every time, must slightly be guided and supported;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3-5 times using Rotary Evaporators, centrifuge successively, supernatant
Spray drying or freeze-drying, sieving, ultraviolet sterilization, packaging is to get to 30KDa above section cocoon sericin protein.
Purposes of the cocoon sericin protein that the method obtains in the drug for preparing antibacterium bioactivity.
Purposes of the cocoon sericin protein that the method obtains in the food for preparing antimycotic bioactivity.
Purposes of the cocoon sericin protein that the method obtains in cosmetics.
Purposes of the cocoon sericin protein that the method obtains in preparing food additives.
A kind of ultrafiltration membrane preparation method of cocoon sericin protein of the present invention, this method by ultrafiltration membrane desalination and
Glue protein matter is concentrated, is a green, high yield, height that is efficient and being easy to a kind of protein of silkworm cocoon (glue protein) that amplification is implemented
Imitate preparation method;What this method obtained silk gum has multiple biological activities and pharmacological action, as antibacterial and its digestive function, suppression
Ultraviolet radiation processed, hypoglycemic effect inhibit tyrosinase activity the natural material of isoreactivity ingredient, and protein content reaches
2204.1 μ g/mL, 88.1%.Pass through the SDS-PAAG electrophoresis showeds of sericine:Sericine contains molecular weight distribution
Within 14.4KDa-66KDa, mainly in 20KDa, 27KDa, 35KDa, 45KDa or so, cosmetics, food and doctor can be used in
The raw material of medicine, removal fibroin makes silk gloss in silk, and the silk gum after removal, which is waste, becomes a kind of high value addition product.
Description of the drawings
Fig. 1 be sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the present invention-silver staining chromatic graph, 13% separation gel, 4%
Glue is concentrated, wherein A is protein molecular weight standard, 66KDa, bovine serum albumin(BSA) .45KDa, ovalbumin .35KDa, pig stomach egg
White enzyme .27KDa, phosphotriose isomerase .20KDa, trypsin inhibitor .14.4KDa, lysozyme .9.5KDa, parathyroid gland
Hormone .6.5KD, Aprotinin, water body glue protein (upper ocean buffer solution), B is sample protein band (silver staining after example 1 is lyophilized
Color);
Fig. 2 is sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the present invention-coomassie brilliant blue staining figure, and wherein A is
Protein molecular weight standard, 66KDa, bovine serum albumin(BSA) 45KDa, ovalbumin 35KDa, porcine pepsin 27KDa, phosphoric acid third
Sugared isomerase 20KDa, trypsin inhibitor 14.4KDa, lysozyme 9.5KDa, parathyroid hormone 6.5KD, Aprotinin, water
Body glue protein (upper ocean buffer solution), B are 1 sample of example, and C is that 3KD concentrates position, and D is that 3KD penetrates position.
Specific implementation mode
Embodiment 1
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, the silk cocoon 100g that weighs that treated, with being dried after petroleum ether degreasing 48h, then again with soaked in absolute ethyl alcohol for 24 hours,
Small molecule ingredient and pigment are removed, is dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, a concentration of 0.05% sodium carbonate liquor refluxing extraction 4 times, every time
20 minutes, obtain crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dried, sieving, ultraviolet sterilization, packaging, you can obtain 3KD above section cocoon sericin protein powders 19.6g.
Embodiment 2
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape.
B, it is dried after the precision silk cocoon 100g that weighs that treated, petroleum ether degreasing 48h, then uses soaked in absolute ethyl alcohol again
Small molecule ingredient and pigment are removed for 24 hours, are dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, a concentration of 0.1% sodium carbonate liquor refluxing extraction 3 times, every time
20 minutes, obtain crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 4 times using Rotary Evaporators, centrifuge successively, supernatant liquid cooling
It is lyophilized dry, sieving, ultraviolet sterilization, packaging is to get to 30KDa above section cocoon sericin albumen powders 20.1g.
Embodiment 3
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, precision weighs that treated then silk cocoon 100g uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
For 24 hours, small molecule ingredient and pigment are removed, is dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, a concentration of 0.08% sodium carbonate liquor refluxing extraction 2 times, every time
20 minutes, obtain crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 5 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dried, and sieving, ultraviolet sterilization, packaging is to get to 30KDa above section cocoon sericin albumen powders 17.9g.
Embodiment 4
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, precision weighs that treated then silk cocoon 100g uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
For 24 hours, small molecule ingredient and pigment are removed, is dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, a concentration of 0.07% sodium carbonate liquor refluxing extraction 4 times, every time
20 minutes, obtain crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dry or freeze-drying, sieving, ultraviolet sterilization are packed, you can obtain 30KDa above section glue protein powder 18.1g.
Embodiment 5
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape.
B, then the precision silk cocoon 100g that weighs that treated uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
Small molecule ingredient and pigment are removed for 24 hours, are dried;
C, by step b gained silk cocoons at 100 DEG C, a concentration of 0.1% sodium carbonate liquor refluxing extraction 3 times, 20 points every time
Clock obtains crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 5 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dry or freeze-drying, sieving, ultraviolet sterilization are packed, you can obtain 30KDa above section glue protein powder 21.5g.
Embodiment 6
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, precision weighs that treated then silk cocoon 100g uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
Small molecule ingredient and pigment are removed for 24 hours, are dried;
C, by step b gained silk cocoons at 100 DEG C, a concentration of 0.1% sodium carbonate liquor refluxing extraction 2 times, 20 points every time
Clock obtains crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dry or freeze-drying, sieving, ultraviolet sterilization are packed, you can obtain 30KDa above section glue protein powder 21.9g.
Embodiment 7
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape.
B, then the precision silk cocoon 100g that weighs that treated uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
Small molecule ingredient and pigment are removed for 24 hours, are dried;
C, by step b gained silk cocoons at 100 DEG C, a concentration of 0.07% sodium carbonate liquor refluxing extraction 3 times, 20 points every time
Clock obtains crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dry or freeze-drying, sieving, ultraviolet sterilization are packed, you can obtain 30KDa above section glue protein powder 20g.
Embodiment 8
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, then the precision silk cocoon 100g that weighs that treated uses soaked in absolute ethyl alcohol again with being dried after petroleum ether degreasing 48h
Small molecule ingredient and pigment are removed for 24 hours, are dried;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, a concentration of 0.1% sodium carbonate liquor refluxing extraction 4 times, every time
20 minutes, obtain crude extract;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant,
Obtain glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 4 times using Rotary Evaporators, centrifuge successively, supernatant spray
Mist is dry or freeze-drying, sieving, ultraviolet sterilization are packed, you can obtain 30KDa above section glue protein powder 20.5g.
Embodiment 9
The measurement of protein concentration:
It is measured using bicinchoninic acid (bicinchoninic acid, BCA) method and implements 3KD above section glue obtained by 1-8
Protein concentration in albumen powder;
The drafting of standard curve operates according to BCA protein measurement kit specifications and prepares working solution, every 50 times of volumes
BCA reagent A solution adds the BCA reagent B solutions of 1 times of volume, mix well it is spare, according to table 1 draw kit in protein standard
Product bovine serum albumin, is diluted with water, and shakes up, and each pipe takes 25 μ L in 96 orifice plates respectively, each that 175 μ L BCA working solutions are added, gently
Micro oscillation, mixing incubate 30min in 37 DEG C of insulating boxs of temperature, take out 96 orifice plates, measure absorbance with microplate reader, measure wave
A length of 562nm.It is each do three groups it is parallel, using absorbance as ordinate, protein concentration is that abscissa carries out regressing calculation;
Table 1 is serially diluted bovine serum albumin(BSA) (BSA) standard items
The measurement of sample:
It is accurate to weigh 2.5mg3KD above section glue protein powder samples, 1mL is added and distills water dissolution, 10000 revs/min of centrifugations
2min takes 25 μ L samples solution in 96 orifice plates according to standard curve operating method, each that 175 μ L BCA working solutions are added, slightly
Oscillation, mixing incubate 30min in 37 DEG C of insulating boxs of temperature, take out 96 orifice plates, measure absorbance with microplate reader, measure wavelength
For 562nm, each sample do three groups it is parallel, the content of protein is calculated according to standard curve;
Experimental result:
By Fig. 2 and table 2 it is found that being extracted regardless of solution, using liquid nitrogen grinding to the extract protein of feedstock processing
Content is extract obtained higher than stirring means processing raw material;Using the extract of same material processing, phosphate is utilized
The protein content of buffer solution extraction is higher than to be extracted using sodium chloride;Protein content is reduced after removing salting-in-protein, therefore, profit
Raw material is crushed with liquid nitrogen grinding method, extract obtained protein content highest is extracted with phosphate buffer, up to 2204.1 μ
G/mL accounts for the 88.1% of extract, is shown in Table 2
2 protein extracting ratio of table is the glue protein percentage amounts extracted per 100g silk cocoons
Experiment condition | Protein extracting ratio | Protein content | |
Example 1 | 0.05% sodium carbonate, 3KD, 48h | 19.6% | 0.88mg/ml |
Example 2 | 0.05% sodium carbonate, 3KD, 72h | 20.1% | 0.78mg/ml |
Example 3 | 0.05% sodium carbonate, 8KD, 48h | 17.9% | 0.81mg/ml |
Example 4 | 0.05% sodium carbonate, 8KD, 72h | 18.1% | 0.59mg/ml |
Example 5 | 0.1% sodium carbonate, 3KD, 48h | 21.5% | 0.63mg/ml |
Example 6 | 0.1% sodium carbonate, 3KD, 72h | 21.9% | 0.61mg/ml |
Example 7 | 0.1% sodium carbonate, 8KD, 48h | 20.0% | 0.59mg/ml |
Example 8 | 0.1% sodium carbonate, 8KD, 72h | 20.5% | 0.68mg/ml |
Embodiment 10
SDS-PAAG collection of illustrative plates freezes 1 protein of embodiment using sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Dry-eye disease (Fig. 1), ultrafiltration membrane ultrafiltration upper bit and non-ultrafiltration solution (Fig. 2) carry out protein classes determination:
Preparation of reagents and sample treatment:
Sample buffer:0.5mL trishydroxymethylaminomethanes and mixed in hydrochloric acid buffer solution (0.5moL/L pH=6.8),
10% lauryl sodium sulfate, 1mL glycerine, the beta -mercaptoethanol of 0.5mL, the water of 1mL, the micro bromophenol blue of 2mL;
The processing of sample:Precision weighs 3 milligrams of sample and is dissolved in 1mL distilled water, shakes up, and is mixed in equal volume with sample buffer
It closes, water proof boils 10 minutes, 10000 revs/min, centrifuges 5 minutes;
Electrode buffer:By 15.1 grams of trishydroxymethylaminomethanes, 10% dodecyl sulphate of 94 grams of glycine, 50mL
Sodium dissolves and is settled to 4L;
Sodium dodecyl sulfate-polypropylene acrylamide gel storing liquid:30% (W/V) extra-pure grade acrylamide, 0.8% (W/
V) N, N- methylene bisacrylamide;
The separation gel of 12% sodium dodecyl sulfate-polypropylene acrylamide gel is prepared:30% (the V/ of H2O, 4mL of 3.3mL
V) sodium dodecyl sulfate-polypropylene acrylamide gel storing liquid, 2.5mL 1.5moL/L trishydroxymethylaminomethane and hydrochloric acid
Cocktail buffer (pH=8.8), 10% (W/V) lauryl sodium sulfate of 0.1mL, 0.1mL 10% (W/V) ammonium persulfate,
The tetramethylethylenediamine of 0.006mL;
The spacer gel of 5% sodium dodecyl sulfate-polypropylene acrylamide gel is prepared:The 30% of H2O, 0.17mL of 0.68mL
(V/V) sodium dodecyl sulfate-polypropylene acrylamide gel storing liquid, 0.13mL trishydroxymethylaminomethanes and mixed in hydrochloric acid buffer
10% (W/V) persulfuric acid of liquid (0.5moL/L pH=6.8), 10% (W/V) lauryl sodium sulfate of 0.01mL, 0.01mL
The tetramethylethylenediamine of ammonium, 0.001mL;
Deposition condition:75 volts of constant pressure, 40 minutes, then 150 volts of constant pressure, 90 minutes;Fixer:25% isopropanol, 10%
Glacial acetic acid, 65% water;Silver staining (uses this method) to preferably observe protein band after freeze-drying;It impregnates:75ml ethyl alcohol,
17g sodium acetates, 1.25ml25% glutaraldehydes, 0.5g sodium thiosulfate 5H2O dH2250ml 30min are added to after O dissolvings;Drift
It washes:Use dH2O is rinsed 3 times 5min/ times;Silver staining:0.25g silver nitrates, 50ul formaldehyde add to 250ml 20min with dH2O;Colour developing:
6.25g sodium carbonate, 25ul formaldehyde, uses dH2O adds to 250ml 2-10min opsin bands;Show dark-brown;It terminates:
3.65gEDTA-Na2·2H2O uses dH2O adds to 250ml 10min;Rinsing:Use dH2O is rinsed 3 times 5min/ times;It preserves:25ml
Glycerine dH2O adds to 250ml 30min taking-ups and dries;
Experimental result:There are many containing in 1 Silver stain of embodiment in sodium dodecyl sulfate-polyacrylamide gel electrophoresis figure
Activated protein, wherein sericine contain molecular weight distribution within 14.4KDa-66KDa.
Implementation column 11
Silk-fibroin amino acid analysis
This method is enlightening equine skill application experiment room respectively using phenyl isothiocyanate, 2,4- dinitrofluorobenzene as derivating agent
Protein hydrolyzate and free amine group acid injection are derived, then detached using Diamonsil AAA columns, it can
Meet the analysis of 19 kinds of natural amino acids, each component separating degree is higher, quantitative result is accurate and stablizes;
Reagent:0.1% phenol+6mol/l HCl/water solution:It weighs 0.1g phenol and is placed in 100ml volumetric flasks, 50ml is added
Concentrated hydrochloric acid (36%-38%, molar concentration are about 12mol/l), then plus water is settled to 100ml, 0.1mol/l HCl/water solution:
8.3ml concentrated hydrochloric acids are measured, are then settled to 1000ml amino acid stocks with pure water:A certain amount of amino acid standard is weighed, is used
0.1mol/l HCl/water solution dissolves, cystine 0.01mol/l, tyrosine 0.02mol/l, other amino acid are
0.05mol/l;
Amino acid uses liquid:Storing solution 0.1mol/l HCl/water solution is diluted, obtains a concentration of 0.002mol/l's
Amino acid list mark and mixed mark (cystine concentration 0.001mol/l) are used for PITC derivatization methods;By storing solution 0.1mol/l HCl
Aqueous solution dilutes, and obtains the amino acid list mark of a concentration of 0.0005mol/l and mixed mark (cystine concentration 0.00025mol/l)
For DNFB derivatization methods;
Nor-leucine internal standard solution:Using nor-leucine as internal standard compound.A certain amount of nor-leucine is weighed, 0.1mol/L is dissolved in
HCl/water solution obtains the nor-leucine internal standard solution of 0.02mol/L;
Phenyl isothiocyanate solution:250 μ l phenyl isothiocyanates are settled to 10ml with acetonitrile, obtain the different sulphur of 0.2mol/L
Phenyl-cyanate solution;
Triethylamine solution:1.4ml triethylamines are settled to 10ml with acetonitrile, obtain 1.0mol/L triethylamine solutions 1%
DNFB acetonitrile solutions:0.5ml 2,4-dinitrofluorobenzene (DNFB) is dissolved in 50ml acetonitriles, and volume fraction 1% is approximately equal to
0.08mol/l;0.1mol/L Na2B4O7 aqueous solutions:1.91g Na2B4O710H2O are weighed, are dissolved with 50ml pure water, solution
PH is about 9.50;
Sample treatment:Protein hydrolyzes (acid-hydrolysis method):Protein example (feed, milk powder, animal tissue etc.):For dynamic
Object sample, crushed after being dried is at fine particle;For feed, it is ground into fine particle;Milk powder is not required to pre-process, and sample is placed in
20m spiral covers (liner PTFE dottle pins) narrow-mouthed bottle is added 0.1% phenol+6mol/l HCl/water solution of 10ml, screws lid, vibrates
Mixing is reacted for 24 hours at 110 DEG C of temperature, and reaction finishes, and reaction solution is transferred completely into 100ml cucurbits, subtracts at 75 DEG C of temperature
Pressure distillation is done to close, to remove HCl remaining in reaction solution, is then dissolved in three times with 12ml 0.1mol/L HCl/waters solution
Residue, and 20ml glass tool plug scale test tubes are transferred to, then it is settled to 20ml with pure water, it waits deriving;
Injection processing:When using PITC derivatization methods, sample is diluted to 4g/l with 0.1mol/l HCl/water solution;When
When deriving using DNFB, sample is diluted to 2g/L with 0.1mol/L HCl/water solution;
The analysis condition of amino acid-PITC derivatives:
Chromatographic column:Diamonsil AAA amino acid analysis columns, 250 × 4.6mm, 5 μm of (CAT#:99751);
Guard column:EasyGuard C18,10 × 4.0mm, 5 μm (CAT#:6201);
Mobile phase A:0.05mol/l aqueous sodium acetate solutions (it is 6.50 ± 0.05 that glacial acetic acid, which adjusts pH value);
Mobile phase B:Methanol:Acetonitrile:Water=20:60:20 (volume ratios);
Flow velocity:1.0ml/min, column temperature:35℃;Sampling volume:10 μ l, Gradient program;Time/min 0,39,40,45,
46,60;Mobile phase A/%95,52,0,0,95,95 Mobile phase Bs/%5,48,100,100,5,5;
3 Gradient program of table
Time/min | 0 | 39 | 40 | 45 | 46 | 60 |
Mobile phase A/% | 95 | 52 | 0 | 0 | 95 | 95 |
Mobile phase B/% | 5 | 48 | 100 | 100 | 5 | 5 |
Experimental result is shown in Table 4:
4 glue protein aminoacid ingredient of table and assay result
Embodiment 13
Determination of elemental analysis:
The method of carbon, hydrogen, nitrogen and sulphur or oxygen (C, H, N, S/O) constituent content in organic matter is measured with elemental analyser, is fitted
| || micro type elemental analyser for adsorbing separation and chromatographic isolation formula;
Method And Principle
Carbon, hydrogen, nitrogen and sulphur in organic matter or oxygen (C, H, N, S/O) element, after catalysis oxidation (or cracking)-reduction
It is transformed into carbon dioxide, vapor, nitrogen and sulfur dioxide or carbon monoxide (CO2, H2O, N2, SO2/CO) respectively.Then exist
Under the promotion of carrier gas, each component of method sequentially determining is examined with adsorbing separation-thermal conductivity difference;Or mixed gas is detached with chromatography
Afterwards, the response signal value of component is measured respectively with thermal conductivity detector (TCD) or infrared absorption detector.According to the signal value of component (or color
Spectrum peak) and corresponding element sensitivity (or correction) factor K value, calculate separately the content of various elements in sample;
Reagent and material:
Standard substance:Benzoic acid, antifebrin, cyclohexanone -2,4- dinitrophenylhydrazone, chlorination S- benzyl thiocarbamides, p-aminophenyl
Sulfonic acid;
ω × 102 of carrier gas (helium or argon gas) purity >=99.99, moisture≤10-6g/L;
ω × 102 of oxygen purity >=99.99, moisture≤10-6g/L;
Instrument forms:Instrument is mainly by filling the oxidation tube of oxidation catalyst and filling with reduction tube and the heating of reducing agent
At stove, separation and the separation of detection oxidation (or cracking)-reduzate CO2, H2O, N2 and SO2 or CO and detecting system, data
Reason machine or microcomputer workstation, micro autoelectrinic balance (see appendix A) and groups of printers at;
Sample:Sample should be free from the homogeneous solid particle or liquid of adsorption moisture, volatile samples low-melting alloy
Container sealing weighs, and corrosive liquids is weighed with low-melting glass capillary seal, and when oxidation should have explosion precaution;
K values measure:Standard sample 1mg-3mg is weighed, 0.001mg is accurate to, by sample evaluation procedures METHOD FOR CONTINUOUS DETERMINATION two
Above standard sample, until result reaches the specified value of instrumentation regulation;
K values calculate:The K values of i elements press (1-a) formula (adsorption separation method) or (1-b) formula (chromatographic isolation in standard sample
Method) it calculates:
The signal value of μ V-standard sample i element unit quality in formula;M-standard sample quality;W-standard sample i members
Plain true value percentage;
M-standard sample quality in formula;W-standard sample i element true value percentages;A-standard sample i elements peak face
Product;The K values of Ki=i (C, H, N and S or O) element in formula (1) and formula (2);
Sample measures:Sample 1mg-3mg is weighed, 0.001mg is accurate to, by sample evaluation procedures METHOD FOR CONTINUOUS DETERMINATION 2 or more
Sample, until result reaches the requirement of analytical error;
Experimental result is shown in Table 5:
5 glue protein constituent content of table
Weight | C% | H% | S% | N% |
1mg | 41.01 | 5.922 | 0.280 | 14.73 |
Embodiment 14
Antioxidant activity is tested:
Experiment reagent:(2,2 '-hexichol are for bitter taste by DPPH (2,2-diphenyl-1-picrylhydrazyl hydrate)
Acyl group phenylhydrazine), it is a kind of lipid free radical centered on nitrogen of stabilization, contains 3 phenyl ring, methanol or ethyl alcohol in structure
Solution is in purple, has strong absorption near 517nm, in the presence of having the antioxidant of hydrogen supply capacity, color can be by dark purple discoloration
It is light yellow[4], the absorbance at 517nm becomes smaller.The degree that its absorbance becomes smaller is proportionate with the ability for removing free radical
Property, thus can be used for detecting antioxidant activity;
Experimental method:
Prepare 6 × 10-5M DPPH (Sigma) methanol solution:1.2mg DPPH are weighed, 500 μ L methanol solutions is dissolved in, is made into
100 × liquid is preserved, it is placed in 4 DEG C of refrigerators of temperature and preserves, answered at working solution, DPPH working solutions with after 100 times of methanol dilution using preceding
It is kept in dark place as possible, it is now with the current;
Be dissolved in ultra-pure water polypeptide sample be added DPPH working solutions, system be 100 μ L, room temperature avoid light place 30min, in
Light absorption is detected at 517nm, free radical is reacted in the reduction relative to blank control (with the ultra-pure water of sample same volume) light absorption
Removing situation, each concentration samples do 3 it is parallel, ultraviolet specrophotometer return to zero when use methanol;
DPPH free radical scavenging activities are calculated by the following formula:
Clearance rate=[(A0-A1)/A0] × 100
Wherein, A0 is blank control absorbance value, and A1 is that sample sets absorbance value is added
Experimental result is shown in Table 6
6 glue protein of table removes the capability result table of free radical
As can be seen from the table:The cocoon sericin protein that the present invention obtains has the activity for removing free radical.
Claims (5)
1. a kind of ultrafiltration membrane preparation method of cocoon sericin protein, it is characterised in that follow these steps to carry out:
A, the pre-treatment of silk cocoon:The filiform on silk cocoon surface is peelled off, remaining impurity in silk cocoon is removed, is cut into tiny item
Shape;
B, the silk cocoon 100g that weighs that treated, with being dried after petroleum ether degreasing 48h, then again with soaked in absolute ethyl alcohol for 24 hours, remove
Small molecule ingredient and pigment, dry;
C, by silk cocoon obtained by step b in 100 DEG C of temperature, the sodium carbonate liquor refluxing extraction of a concentration of 0.05-0.1% 2-4 times, often
It secondary 20 minutes, must slightly be guided and supported;
D, after the crude extract concentration obtained step c, under 12000 revs/min, 4 DEG C of centrifugation 10min of temperature take supernatant, obtain
Glue protein extracting solution;
E, the glue protein crude extract for obtaining step d uses molecular cut off for the ultrafiltration membrane again of 30KDa, rotating speed
3000rmp/min collects top, obtains glue protein extraction solution;
F, step e is obtained into glue protein and extracts solution, be concentrated into 3-5 times using Rotary Evaporators, centrifuge successively, supernatant spraying
Dry or freeze-drying, sieving, ultraviolet sterilization, packaging is to get to 30KDa above section cocoon sericin protein.
2. use of the cocoon sericin protein that method as described in claim 1 obtains in the drug for preparing antibacterium bioactivity
On the way.
3. use of the cocoon sericin protein that method as described in claim 1 obtains in the food for preparing antimycotic bioactivity
On the way.
4. purposes of the cocoon sericin protein that method as described in claim 1 obtains in cosmetics.
5. purposes of the cocoon sericin protein that method as described in claim 1 obtains in preparing food additives.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810686464.0A CN108743446A (en) | 2018-06-28 | 2018-06-28 | A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810686464.0A CN108743446A (en) | 2018-06-28 | 2018-06-28 | A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application |
Publications (1)
Publication Number | Publication Date |
---|---|
CN108743446A true CN108743446A (en) | 2018-11-06 |
Family
ID=63974373
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810686464.0A Pending CN108743446A (en) | 2018-06-28 | 2018-06-28 | A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108743446A (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109984857A (en) * | 2019-04-10 | 2019-07-09 | 华中科技大学同济医学院附属协和医院 | A kind of method for building up of precision peritoneal adhesion animal model and application |
CN114404568A (en) * | 2022-01-16 | 2022-04-29 | 重庆理工大学 | Sericin polypeptide injection and application thereof |
CN114989285A (en) * | 2022-06-16 | 2022-09-02 | 华南农业大学 | Multifunctional mulberry silk collagen and application thereof in biomedical materials |
CN115531284A (en) * | 2022-09-15 | 2022-12-30 | 江苏华佳丝绸股份有限公司 | Preparation method of silk mask containing sericin |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103290085A (en) * | 2013-05-13 | 2013-09-11 | 湖州新天丝生物技术有限公司 | Silk protein powder and preparation method thereof |
CN103830119A (en) * | 2014-03-11 | 2014-06-04 | 安徽省农业科学院蚕桑研究所 | Preparation method of toning lotion taking natural sericin as matrix |
CN107937460A (en) * | 2016-10-13 | 2018-04-20 | 宜宾屏山辉瑞油脂有限公司 | A kind of preparation method for hydrolyzing sericin peptide taken |
-
2018
- 2018-06-28 CN CN201810686464.0A patent/CN108743446A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103290085A (en) * | 2013-05-13 | 2013-09-11 | 湖州新天丝生物技术有限公司 | Silk protein powder and preparation method thereof |
CN103830119A (en) * | 2014-03-11 | 2014-06-04 | 安徽省农业科学院蚕桑研究所 | Preparation method of toning lotion taking natural sericin as matrix |
CN107937460A (en) * | 2016-10-13 | 2018-04-20 | 宜宾屏山辉瑞油脂有限公司 | A kind of preparation method for hydrolyzing sericin peptide taken |
Non-Patent Citations (2)
Title |
---|
王菊生,等: "《染整工艺原理》", 30 June 1990, 纺织工业出版 * |
蒋猷龙: "《家蚕的生活和习性》", 30 November 1980, 农业出版社 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109984857A (en) * | 2019-04-10 | 2019-07-09 | 华中科技大学同济医学院附属协和医院 | A kind of method for building up of precision peritoneal adhesion animal model and application |
CN114404568A (en) * | 2022-01-16 | 2022-04-29 | 重庆理工大学 | Sericin polypeptide injection and application thereof |
CN114404568B (en) * | 2022-01-16 | 2023-12-26 | 重庆理工大学 | Sericin polypeptide injection preparation and application thereof |
CN114989285A (en) * | 2022-06-16 | 2022-09-02 | 华南农业大学 | Multifunctional mulberry silk collagen and application thereof in biomedical materials |
CN114989285B (en) * | 2022-06-16 | 2024-01-30 | 华南农业大学 | Multifunctional mulberry silk sericin and application thereof in biomedical materials |
CN115531284A (en) * | 2022-09-15 | 2022-12-30 | 江苏华佳丝绸股份有限公司 | Preparation method of silk mask containing sericin |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108743446A (en) | A kind of ultrafiltration membrane preparation method of cocoon sericin protein and its application | |
Silva et al. | Extraction of collagen/gelatin from the marine demosponge Chondrosia reniformis (Nardo, 1847) using water acidified with carbon dioxide–process optimization | |
CN104450839B (en) | The preparation method of the rice bran protein peptide with ACE inhibitory activity | |
CN109400678A (en) | A kind of anti-oxidant and DPP-IV inhibitory activity peptide in stichopus japonicus source | |
CN104672316B (en) | A kind of silk fibroin protein solution prepares and identification method | |
CN108456244A (en) | Corn antioxidant active peptide and preparation method thereof | |
JP6253047B2 (en) | Proteoglycan and cosmetics | |
CN110317848A (en) | A kind of preparation method of collagen peptide | |
CN109680031A (en) | A kind of extracting method of Elastin peptide | |
CN108611391A (en) | A kind of method of modifying of collagen from black sea cucumbers from East China Sea oligopeptide and its application | |
CN115960165B (en) | Selenium-enriched ACE (angiotensin converting enzyme) inhibitory peptide derived from moringa leaves and application thereof | |
CN104177477A (en) | Fish anti-oxidation active peptide and preparation method thereof | |
CN112625088A (en) | Preparation method and application of mussel ACE inhibitory peptide | |
CN104558115A (en) | Antioxidant polypeptide with Raja porosa meat protein as well as preparation method and application of antioxidant polypeptide | |
CN108456707A (en) | A kind of preparation method of pupa albumen anti-oxidation peptide and the application of pupa albumen anti-oxidation peptide | |
CN112501229B (en) | Production process of bovine bone collagen peptide | |
CN105131099B (en) | A method of sericin is prepared from tussah degumming of silk industrial wastewater | |
CN103882085A (en) | Combined extraction preparation method of complex polypeptide | |
CN117281885A (en) | Application of selenium-enriched yam glycoprotein as immunomodulating drug | |
CN107082796B (en) | Method for purifying small molecular polypeptide in protein zymolyte | |
CN109354600A (en) | A kind of preparation method of the novel Alaska pollock multifunctional polypeptide of taurine modification | |
CN110655553B (en) | ACE inhibitory peptide derived from sesame, preparation method and application thereof in preparation of antihypertensive drugs | |
CN108191958A (en) | A kind of anti-oxidant hexapeptide of flaxseed meal and preparation method and application | |
CN108059652B (en) | A kind of grifola frondosus selenium chelating peptide prepared using proteolytic cleavage | |
CN111388505A (en) | Preparation method of animal placenta transfer factor |
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 | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20181106 |
|
WD01 | Invention patent application deemed withdrawn after publication |