CN112851764B - 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 - Google Patents
一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 Download PDFInfo
- Publication number
- CN112851764B CN112851764B CN202110260036.3A CN202110260036A CN112851764B CN 112851764 B CN112851764 B CN 112851764B CN 202110260036 A CN202110260036 A CN 202110260036A CN 112851764 B CN112851764 B CN 112851764B
- Authority
- CN
- China
- Prior art keywords
- leu
- lrp
- regium
- antioxidant peptide
- pleurotus tuber
- 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.)
- Active
Links
- 101800000068 Antioxidant peptide Proteins 0.000 title claims abstract description 31
- 241000149440 Lentinus tuber-regium Species 0.000 title abstract description 27
- 102000004169 proteins and genes Human genes 0.000 title abstract description 8
- 108090000623 proteins and genes Proteins 0.000 title abstract description 8
- 235000013399 edible fruits Nutrition 0.000 title description 3
- 239000003814 drug Substances 0.000 claims abstract description 13
- 230000006907 apoptotic process Effects 0.000 claims abstract description 11
- DIVDFFZHCJEHGG-UHFFFAOYSA-N oxidopamine Chemical compound NCCC1=CC(O)=C(O)C=C1O DIVDFFZHCJEHGG-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims description 6
- 210000002569 neuron Anatomy 0.000 claims description 4
- 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 claims description 2
- 230000002000 scavenging effect Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 32
- 229920001184 polypeptide Polymers 0.000 abstract description 31
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 31
- 150000001413 amino acids Chemical class 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 9
- 239000000203 mixture Substances 0.000 abstract description 8
- 229940079593 drug Drugs 0.000 abstract description 6
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 4
- 230000002441 reversible effect Effects 0.000 abstract description 3
- 238000002270 exclusion chromatography Methods 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 206010034010 Parkinsonism Diseases 0.000 abstract 1
- 238000002514 liquid chromatography mass spectrum Methods 0.000 abstract 1
- 210000005036 nerve Anatomy 0.000 abstract 1
- 210000000653 nervous system Anatomy 0.000 abstract 1
- 230000001590 oxidative effect Effects 0.000 abstract 1
- 239000003223 protective agent Substances 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 27
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 27
- 210000004027 cell Anatomy 0.000 description 19
- 208000018737 Parkinson disease Diseases 0.000 description 17
- 239000011347 resin Substances 0.000 description 11
- 229920005989 resin Polymers 0.000 description 11
- 101000579904 Homo sapiens Leucine-rich repeat-containing protein 3B Proteins 0.000 description 10
- 102100027492 Leucine-rich repeat-containing protein 3B Human genes 0.000 description 10
- 239000000843 powder Substances 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 8
- 239000000284 extract Substances 0.000 description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 6
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 6
- 230000036542 oxidative stress Effects 0.000 description 6
- 235000018102 proteins Nutrition 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- 230000007760 free radical scavenging Effects 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000004090 neuroprotective agent Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000001640 apoptogenic effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 229960003638 dopamine Drugs 0.000 description 3
- 210000005064 dopaminergic neuron Anatomy 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 238000004007 reversed phase HPLC Methods 0.000 description 3
- 238000010532 solid phase synthesis reaction Methods 0.000 description 3
- 239000008223 sterile water Substances 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- ZGYICYBLPGRURT-UHFFFAOYSA-N tri(propan-2-yl)silicon Chemical compound CC(C)[Si](C(C)C)C(C)C ZGYICYBLPGRURT-UHFFFAOYSA-N 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 241001573498 Compacta Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 2
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 2
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 2
- 239000012505 Superdex™ Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 238000009098 adjuvant therapy Methods 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000002983 circular dichroism Methods 0.000 description 2
- 238000001142 circular dichroism spectrum Methods 0.000 description 2
- 239000002173 cutting fluid Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000002523 gelfiltration Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 230000004770 neurodegeneration Effects 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- 230000004112 neuroprotection Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000001542 size-exclusion chromatography Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 210000003523 substantia nigra Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 229910001868 water Inorganic materials 0.000 description 2
- CBPJQFCAFFNICX-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-methylpentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(C)C)C(O)=O)C3=CC=CC=C3C2=C1 CBPJQFCAFFNICX-IBGZPJMESA-N 0.000 description 1
- VIYKYVYAKVNDPS-HKGPVOKGSA-N (2s)-2-azanyl-3-[3,4-bis(oxidanyl)phenyl]propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1.OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 VIYKYVYAKVNDPS-HKGPVOKGSA-N 0.000 description 1
- LZOLWEQBVPVDPR-VLIAUNLRSA-N (2s,3r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]butanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H]([C@H](OC(C)(C)C)C)C(O)=O)C3=CC=CC=C3C2=C1 LZOLWEQBVPVDPR-VLIAUNLRSA-N 0.000 description 1
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 description 1
- 102000000412 Annexin Human genes 0.000 description 1
- 108050008874 Annexin Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- WVLZTXGTNGHPBO-SRVKXCTJSA-N Cys-Leu-Leu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O WVLZTXGTNGHPBO-SRVKXCTJSA-N 0.000 description 1
- 208000012661 Dyskinesia Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- YFBBUHJJUXXZOF-UWVGGRQHSA-N Leu-Gly-Pro Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O YFBBUHJJUXXZOF-UWVGGRQHSA-N 0.000 description 1
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 1
- 241000208566 Lignosus rhinocerotis Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000222350 Pleurotus Species 0.000 description 1
- JDJMFMVVJHLWDP-UNQGMJICSA-N Pro-Thr-Phe Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O JDJMFMVVJHLWDP-UNQGMJICSA-N 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 241000566137 Sagittarius Species 0.000 description 1
- BVOVIGCHYNFJBZ-JXUBOQSCSA-N Thr-Leu-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(O)=O BVOVIGCHYNFJBZ-JXUBOQSCSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 102000003802 alpha-Synuclein Human genes 0.000 description 1
- 108090000185 alpha-Synuclein Proteins 0.000 description 1
- 150000001371 alpha-amino acids Chemical class 0.000 description 1
- 235000008206 alpha-amino acids Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000013528 artificial neural network Methods 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000001120 cytoprotective effect Effects 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- WJJMNDUMQPNECX-UHFFFAOYSA-N dipicolinic acid Chemical compound OC(=O)C1=CC=CC(C(O)=O)=N1 WJJMNDUMQPNECX-UHFFFAOYSA-N 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000011866 long-term treatment Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000009125 negative feedback regulation Effects 0.000 description 1
- 230000000324 neuroprotective effect Effects 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 238000005025 nuclear technology Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- 230000007065 protein hydrolysis Effects 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 238000009256 replacement therapy Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- 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
- 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
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Psychiatry (AREA)
- Hospice & Palliative Care (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用,通过分子排阻色谱和反向高效液相色谱法分离纯化LRP‑15,通过液相色谱‑质谱连用测定LRP‑15的氨基酸组成。该抗氧化肽包含以下序列:Thr‑Leu‑Ala‑Pro‑Thr‑Phe‑Leu‑Ser‑Ser‑Leu‑Gly‑Pro‑Cys‑Leu‑Leu。验证并解释LRP‑15对6‑OHDA诱导的PC12细胞的凋亡的缓解作用。获得的活性多肽是一种优良的神经保护剂,具有减缓氧化压力对神经系统损伤的功能,可用于制备预防和辅助治疗帕金森综合症的药物或保健食品中。
Description
技术领域
本发明属于生物小分子活性肽领域,涉及一种来源于虎奶菇子实体蛋白的抗氧化肽。
背景技术
帕金森病(Parkinson's disease,PD)是人类第二大神经退行性疾病,65岁以上人群中患病率为2-3%。PD的病理特征可概括为黑质致密区大量多巴胺能神经元丢失,进而导致黑质和纹状体多巴胺生物合成减少。PD的致病机理还不明确,研究证实,环境因素、遗传因素、氧化应激等均可能参与PD多巴胺能神经元的变性死亡过程。其中,氧化应激是导致神经元退化变性的一个重要原因,一方面,大脑是人类正常的生理活动中耗氧量最大的器官,但脑内的抗氧化酶相对缺乏,使大脑非常容易受到氧化应激损伤;另一方面,早期研究表明,黑质致密区多巴胺能神经元线粒体氧化应激的增加可触发多巴胺依赖性毒性级联反应,导致溶酶体功能障碍和α-突触核蛋白的积累,这是PD的两个主要病理特征。目前,医学上大多采用经典的多巴胺替代疗法来治疗PD,如L-3,4-二羟基苯丙氨酸(L-DOPA)的补充。但越来越多的研究发现,这一经典的疗法存在较大的副作用,临床观察发现,长周期的L-DOPA治疗,有一定概率导致PD患者的运动障碍。此外,有学者已经从长周期的L-DOPA治疗的PD病人的尿液中分离到6-OHDA,而6-OHDA的存在会使得神经细胞中活性氧含量升高,进而会引发细胞凋亡,可能对PD的治疗起到负反馈调节作用。因此,寻找新的神经保护剂,从缓解氧化压力入手,是预防治疗PD的潜在途径之一。
多肽(polypeptide)是α-氨基酸以肽键连接在一起而形成的化合物,它也是蛋白质水解的中间产物,是源于蛋白质的多功能化合物,且通常情况下营养和生理功能优于蛋白质(大分子蛋白质)和氨基酸。目前,多肽类药物的研究进展迅速,获得的多肽药物主要包括多肽疫苗、抗肿瘤多肽、抗病毒多肽等,多肽药物与一般的有机小分子药物相比,具有生物活性强、用药剂量小、毒副作用低和疗效显著等突出特点。
虎奶菇(Lignosus rhinocerotis)是原产于马来西亚的一种珍稀食药用菌,在马来西亚传统医学中已有上百年的应用历史。虎奶菇的子实体和菌核均可入药,用以治疗食欲不振,身体疲劳,咳嗽,哮喘等疾病,有助于;现代医学确认,虎奶菇子实体及菌核提取物能够促进小鼠大脑神经网络的形成,具有改善认知的功效。但是,具体是提取物中的哪种化合物起作用尚不明了。
发明内容
本发明所要解决的技术问题为:如何提供一种神经保护剂,从而缓解传统PD疗法导致的氧化压力。
本发明的技术方案为:一种虎奶菇源抗氧化肽LRP15,该抗氧化肽LRP15的氨基酸组成为Thr-Leu-Ala-Pro-Thr-Phe-Leu-Ser-Ser-Leu-Gly-Pro-Cys-Leu-Leu,如SEQ IDNo.1所示,相对分子质量为1548Da。
本发明的抗氧化肽LRP15在制备治疗帕金森病的药物上的应用。
优选地,所述药物是以有效量的抗氧化肽LRP-15为活性成分,加上医学上可接受的辅料或者辅助性成分制备而成的制剂。以体重计,抗氧化肽LRP15有效量为0.1-0.2mg/kg。
本发明的抗氧化肽LRP15在制备预防帕金森病的保健食品上的应用。
优选地,所述保健食品是以有效量的抗氧化肽LRP-15为活性成分,加上医学上可接受的辅料或者辅助性成分制备而成的制剂。抗氧化肽LRP15的有效量为2-4mg/天。
本发明的的抗氧化肽LRP15在制备降低6-OHDA导致的神经细胞凋亡的药物或保健食品上的应用。
与现有技术相比,本发明具有以下有益效果:
本发明从虎奶菇子实体中提取并纯化多肽,通过细胞模型验证其神经保护,最终,初筛发现1条多肽LRP-15具有较好的神经保护活性,进一步研究完善了提取方法并鉴定了该多肽的氨基酸序列。LRP-15可以作为神经保护剂用于PD的预防和治疗,有望作为保健品或药品开发利用。
附图说明
图1为分子排阻色谱分离图,A1-A4为分子量由大到小的四个洗脱峰;
图2为组分A2通过RP-HPLC进行分离纯化后色谱图;
图3为LRP-15的圆二色谱;
图4为LRP-15不同浓度下对DPPH自由基的清除活性;
图5为不同浓度的LRP-15对6-OHDA导致的PC12细胞损伤的恢复活力;
图6为LRP-15对PC12细胞凋亡的影响。
具体实施方式
下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为从商业渠道购买得到的。
本申请中,如无特殊说明,%均表示其质量百分含量,即wt%。
实施例1虎奶菇源抗氧化肽的分离纯化及序列鉴定:
称取虎奶菇子实体干品50g,置于中药材粉碎机粉碎后过100目筛网,获得虎奶菇子实体干品粉末。
配置磷酸缓冲盐溶液缓冲液(PBS缓冲液):磷酸二氢钾(KH2PO4)0.27g,磷酸氢二钠(Na2HPO4)1.42g,氯化钠(NaCl)8g,氯化钾(KCl)0.2g,加去离子水约800mL充分搅拌溶解,然后加入浓盐酸调pH至7.4,最后定容到1L。
按照粉末/PBS缓冲液的料液比1:30加样,充分搅拌后置于磁力搅拌器120转搅拌过夜,静置后取上清。上清液通过截留分子量为3500Da的透析袋透析24h,收集透析液,旋转蒸发仪浓缩,获得的浓缩液采用真空冷冻干燥机冷冻干燥,可获得虎奶菇子实体多肽提取物干粉。
取虎奶菇子实体多肽提取物干粉10g,无菌水溶解,配置成浓度为100mg/mL的溶液,上样于Superdex 30Increase凝胶过滤柱,无菌水洗脱,通过自动收集器收集洗脱液,后于215nm下测定洗脱液的吸光值,收集并冻干单一峰,得到虎奶菇子实体多肽复合物。
分子排阻色谱分离结果如图1所示,多肽提取物依据分子量的大小,可以获得4个洗脱峰,分子量由大到小以此标记为A1-4,通过分别收集洗脱峰,测定其自由基清除活性,选择A2组分进行下一步纯化。组分A2通过RP-HPLC进行分离纯化后(图2),获得3个组分,即B1-3。测定自由基清除活性后,选择B2作为虎奶菇多肽,即LRP-15。
取虎奶菇子实体多肽复合物干粉1g并溶解于无菌水,配置成浓度为10mg/mL的溶液,通过反向高效液相色谱纯化,收集单一峰,获得虎奶菇子实体多肽(LRP-15)。色谱条件设置如下:色谱柱选用PE C18柱(150mm×4.6mm),A相:纯净水,B相:含0.1%三氟乙酸的乙腈;进样量设定为30μL;柱温设置为25±5℃;流速选择1.0mL/min;检测波长设定为215nm。洗脱条件:A相:0-8min,99%-97%;8-12min,97%-96%;12-16min,96%-80%;16-20min,80%-99%。
通过液相色谱-质谱(LC-MS-MS)解析LRP-15的氨基酸组成。液相A液为含0.1%甲酸水溶液,B液为0.1%甲酸乙腈溶液。色谱柱以A液平衡。样品由自动进样器上样,再经色谱柱梯度分离,流速为0.3mL/min,柱温70℃。质谱条件:样品用TripleTOF 5600+质谱仪进行质谱分析。分析时长:18min,检测方式:正离子,一级质谱扫描范围:600-3500m/z。该抗氧化肽由15个氨基酸组成,相对分子质量为1532Da。Thr-Leu-Ala-Pro-Thr-Phe-Leu-Ser-Ser-Leu-Gly-Pro-Cys-Leu-Leu(SEQ ID No.1)。
50g虎奶菇子实体粉碎后通过磷酸盐缓冲液提取,3500Da透析袋透析并将所得透析液冻干后共得到多肽提取物干粉9.85g,得率为19.7%。10g多肽提取物经过分子排阻色谱Superdex 30Increase凝胶过滤后,获得多肽复合物冻干粉0.51g,得率为5.1%。1g多肽复合物冻干粉最终通过反向高效液相色谱(RP-HPLC)纯化,获得LRP-15冻干粉0.12g,得率为12%。
实施例2虎奶菇源抗氧化肽LRP-15的二级结构测试
LRP-15的氨基酸组成简单,通过圆二色谱分析确定其二级结构。将LRP-15溶解于水中并制备成浓度为0.25mg/mL的溶液。使用2mm步径的石英比色皿在25℃下进行圆二色谱分析。在190-260nm范围内,以1nm间隔和1nm带宽采集数据。如图3所示,LRP-15的圆二色谱光谱在198nm附近观察到强负带,在220nm附近观察到弱正带。这两个吸收峰显示LRP-15具备不规则二级结构的特征。进一步的统计分析证实,在水溶液中53.9%的LRP-15呈无规卷曲,34.6%的LRP-15呈β-翻转。
实施例3虎奶菇源抗氧化肽的的自由基清除活性
将虎奶菇源抗氧化肽冻干粉(10mg)溶解在10mL去离子水中,稀释成合适浓度(0-1mg/mL)。分别取1mL虎奶菇源抗氧化肽的样品检测液,加入2.0mL 80%乙醇配制的浓度为0.2mM DPPH-溶液,摇匀,静置30min。80%乙醇调零,在517nm处测定样品的吸光值。1.0mL检测液与2.0mL 80%乙醇混匀,波长517nm处的吸光度作对照,以Vc作为阳性对照。DPPH-清除能力(%)=[1-(As-As-blank)/Ac]×100%。式中:Ac代表空白对照(未加样品)吸光度;As代表加入样品溶液后反应体系的吸光度;As-blank代表样品吸光度。
图4显示,虎奶菇源抗氧化肽的具有极好的自由基清除活性,对DPPH自由基清除活性的IC50值约为0.18mg/mL,且具有剂量依赖关系,清除活性与同浓度的阳性对照Vc接近,某些浓度下甚至优于Vc。
实施例4虎奶菇源抗氧化肽LRP-15的细胞保护作用
PC12细胞用含5%胎牛血清和10%马血清的1640培养基培养。取对数期细胞按照3×104个/mL浓度接种于96孔板,每孔100μL。随后,PC12细胞与不同浓度的LRP-15共同孵育6h,移除培养基,加入100μM的6-OHDA处理24h,之后通过MTT法测定PC12细胞的活力。
通过LRP-15的预先孵育,6-OHDA对PC12细胞的损伤作用显著减少(图5)。20μM的LRP-15预处理,使得细胞活力恢复为61.54±2.77%,与未经LRP-15预处理的PC12细胞呈现显著性差异。40μM的LRP-15则可使PC12细胞活力保持在70.16±2.13%。
实施例5虎奶菇源抗氧化肽LRP-15对PC12细胞凋亡的影响
取对数生长期的PC12细胞,经最适浓度的LRP-15预处理后,加入100μM的6-OHDA处理24h,收集细胞,用结合缓冲液调节细胞密度为1×106个/mL,取100μL细胞悬液,加5μLAnnexin/FITC和10μL碘化丙锭溶液,常温避光孵育15min,加入400μL PBS混匀,流式细胞仪分析。
如图6所示,6-OHDA会引起PC12细胞的凋亡,图6A中,早期凋亡的PC12细胞为8.86%,晚期凋亡的PC12细胞则为71.23%,通过LRP-15的预处理,早期凋亡细胞为20.55%,晚期为41.05%,说明LRP-15能在凋亡初期阻断细胞的凋亡,对6-OHDA引起的凋亡具有抑制作用。
实施例6抗氧化肽LRP-15的合成
本发明分离纯化并鉴定出虎奶菇源抗氧化肽LRP-15,受制于虎奶菇子实体的稀少,本发明提供一种固相合成方法合成和生产LRP-15,包括但不限于Fmoc-SPPS方式,BOC-SPPS方式,片段缩合连接方式,从而更高效的获得虎奶菇源抗氧化肽,为该条多肽的进一步利用提供材料。
固相合成所需试剂原料:2-CL树脂,氨基酸(以下用Fmoc-aa-oh表示),二甲基甲酰胺(DMF),二氯甲烷(DCM),甲醇,N,N’-二异丙基碳二亚胺(DIC),N,N-二异丙基乙胺(DIEA),1-羟基苯并三唑(HOBT),三氟乙酸(TFA),三异丙基硅烷(TIS),纯水,EDT,乙醚。
一种固相合成抗氧化肽LRP15的方法,抗氧化肽LRP15的氨基酸组成为Thr-Leu-Ala-Pro-Thr-Phe-Leu-Ser-Ser-Leu-Gly-Pro-Cys-Leu-Leu,包括以下步骤:
a.称量2-CL树脂3g,加入反应器中用20mL的二氯甲烷(DCM)浸泡10min后用N,N-二甲基甲酰胺(DMF)洗涤2次,再用DCM洗涤一次,备用;加入1.2mmol的Fmoc-Thr(tBu)-OH,加入15mL DCM,2mL N,N-二异丙基乙胺(DIEA),反应90min,DCM易挥发,反应过程中需补加DCM。
b.90min后补加4mL甲醇,10mL DCM,封闭反应20min。
c.用DMF洗涤4次,加入哌啶(20%哌啶+80%DMF),脱除Fmoc,反应时间为20min。用DMF洗涤树脂5次,取少量树脂(10-20粒)加入检测管中,加入茚三酮(5g/100mL分析乙醇)2滴,吡啶2滴,100℃加热2min,显色。
d.称取下一个氨基酸Fmoc-Leu-OH(0.9mmol)+HOBT(0.9mmol),加入反应器中,加入15mL DMF,在加入2mL DIC反应1h之后用DMF洗涤4次,取少量树脂检测,无色即可。
e.根据LRP15的氨基酸序列依次更换氨基酸原料,重复c,d步骤直至多肽的偶联结束。
f.把树脂用甲醇抽干后放置于50mL离心管,加切割液(95%TFA+1%H2O+2%EDT+2%TIS)40mL,摇晃切割2h,最后把切割液(过滤走树脂)加入到4个新的50mL离心管中,再加入40mL冰乙醚,摇晃均匀,3000rpm/min离心2min,弃去上层乙醚,收集底部的多肽,得到粗品多肽。
g.通过HPLC法纯化多肽,得到纯度大于95%的多肽,通过真空冷冻干燥机冻干,获得多肽干粉。
以上实验证明,虎奶菇源抗氧化多肽LRP-15,能够缓解6-OHDA引起的细胞凋亡,具有神经保护作用,对神经退行性疾病,特别是帕金森综合症具有一定的预防和辅助治疗作用,是一种优良的神经保护剂,可用于制备预防和辅助治疗帕金森综合症的药物或保健品中。
序列表
<110> 四川省农业科学院生物技术核技术研究所
<120> 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 15
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Thr Leu Ala Pro Thr Phe Leu Ser Ser Leu Gly Pro Cys Leu Leu
1 5 10 15
Claims (4)
1.一种抗氧化肽,其氨基酸序列如SEQ ID No.1所示。
2.权利要求1所述的抗氧化肽在制备降低6-OHDA导致的神经细胞凋亡的药物上的应用。
3.权利要求1所述的抗氧化肽在制备清除自由基的药物上的应用。
4.根据权利要求3所述的应用,其特征在于,所述自由基为DPPH。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110260036.3A CN112851764B (zh) | 2021-03-10 | 2021-03-10 | 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110260036.3A CN112851764B (zh) | 2021-03-10 | 2021-03-10 | 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112851764A CN112851764A (zh) | 2021-05-28 |
CN112851764B true CN112851764B (zh) | 2022-04-22 |
Family
ID=75993851
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110260036.3A Active CN112851764B (zh) | 2021-03-10 | 2021-03-10 | 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112851764B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116621934B (zh) * | 2023-05-27 | 2024-03-29 | 上海市农业科学院 | 来源于草菇的抗皮肤氧化和抑制氧化应激的多肽及其应用 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104277135A (zh) * | 2014-09-23 | 2015-01-14 | 华南农业大学 | 一种虎奶菇多糖及其提取方法 |
CN109762745A (zh) * | 2019-03-11 | 2019-05-17 | 河北医科大学第四医院 | 一种虎奶菇的发酵培养方法以及从培养物中提取活性成分的方法 |
CN109771452B (zh) * | 2019-03-11 | 2021-09-21 | 河北医科大学第四医院 | 虎奶菇提取物在制备治疗肿瘤药物中的应用 |
CN111560052B (zh) * | 2020-05-26 | 2021-04-20 | 四川省农业科学院生物技术核技术研究所 | 一种羊肚菌源抗氧化肽及其制备方法和应用 |
-
2021
- 2021-03-10 CN CN202110260036.3A patent/CN112851764B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN112851764A (zh) | 2021-05-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109400678A (zh) | 一种刺参来源的抗氧化和dpp-iv抑制活性肽 | |
CN109293740B (zh) | 一种牡蛎来源的ace抑制及抗肿瘤活性肽 | |
CN111560052B (zh) | 一种羊肚菌源抗氧化肽及其制备方法和应用 | |
CN110655556A (zh) | 一种免疫调节肽的制备及方法 | |
CN112851764B (zh) | 一种来源于虎奶菇子实体蛋白的抗氧化肽及其应用 | |
CN111057144B (zh) | 一种基因重组胶原寡肽mys-1及其制备方法与应用 | |
CN113845565B (zh) | 一种地龙生物活性小肽及其制备方法和应用 | |
CN105648010B (zh) | 一种激活Nrf2-ARE通路的双髻鲨鱼肉抗氧化肽的制备方法 | |
CN114989248B (zh) | 一种具有抗炎抗氧化活性的忧遁草多肽及其制备方法和应用 | |
CN111269292A (zh) | 具有促进组织修复的家蝇多肽及其制备方法与应用 | |
CN114213506B (zh) | 一种人参来源的抗血管生成及抗肿瘤活性肽及其制备方法和应用 | |
CN107325154B (zh) | 一种具有改善记忆功效的多肽及其分离制备方法和用途 | |
CN115124591A (zh) | 钝顶螺旋藻藻蓝蛋白血管紧张素转化酶抑制肽及其制备方法和应用 | |
CN111187343B (zh) | 一种牡丹类2s白蛋白及其提取方法和应用 | |
CN110655553B (zh) | 一种芝麻来源的ace抑制肽、制备方法及其在制备降血压药物方面的应用 | |
WO2020259109A1 (zh) | 植物生产的epo与转铁蛋白的融合蛋白及其应用 | |
CN109467590B (zh) | 活性肽、重组载体、重组细胞、药物组合物及其制备方法和应用 | |
CN109517034B (zh) | 活性肽、重组载体、重组细胞、药物组合物及其制备方法和应用 | |
CN106699842A (zh) | 一种新的抗炎小分子多肽及其应用 | |
CN114195860B (zh) | 一种轮叶党参来源的抗血管生成及抗肿瘤活性肽及其制备方法和应用 | |
CN109517035B (zh) | 一种西兰花蛋白来源的ace抑制肽、ace抑制肽酶切代谢产物及其制备方法和应用 | |
CN105566446B (zh) | 一种蒲公英籽抗氧化肽及其制备方法与应用 | |
CN109021072A (zh) | 一种红毛藻抗氧化肽及其制备方法 | |
CN108976290A (zh) | 一种红毛藻抗氧化肽的制备方法 | |
CN114656522B (zh) | 一种抗氧化肽及其制备方法和应用 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |