软珊瑚Sarcophyton glaucum中新型细胞毒性寡肽的纯化与鉴定（英文）
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摘要
目的:纯化和鉴定软珊瑚Sarcophyton glaucum蛋白水解物中的细胞毒性肽。创新点:首次从软珊瑚S. glaucum中发现具有癌细胞毒性的寡肽,开发了一种有潜力的新型抗癌药物。方法:通过碱性蛋白酶、胰凝乳蛋白酶、木瓜蛋白酶和胰蛋白酶产生软珊瑚S. glaucum蛋白质水解物,并通过MTT法评估其对人宫颈癌HeL a细胞的毒性。通过膜超滤、凝胶过滤色谱、固相萃取和反相高效液相色谱等技术进一步提纯肽;采用从头测序法(de novo sequencing)进行肽鉴定;根据所鉴定的序列,进一步确定合成肽的细胞毒性。结论:从软珊瑚S. glaucum的木瓜蛋白酶水解物中鉴定出三种新型肽序列:AGAPGG、AERQ和RDTQ(分子量分别为428.45、502.53和518.53 Da)。此三种寡肽具有对HeLa的细胞毒性,其半最大效应浓度(EC50)值各为8.6、4.9和5.6 mmol/L,为抗癌药物5-氟尿嘧啶的3.3、5.8和5.1倍。当测试对非癌细胞Hek293的毒性时,这三种肽仅显示16%、25%和11%的细胞毒性。因此,AERQ、AGAPGG和RDTQ有很大的潜力成为肽类抗癌药物。
Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay on the human cervical cancer(HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction(SPE), and reversed-phase high-performance liquid chromatography(RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ(428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration(EC50) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC50, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs. G
Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay on the human cervical cancer(HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction(SPE), and reversed-phase high-performance liquid chromatography(RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ(428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration(EC50) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC50, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs. G
引文
Abdel-Lateff A,Alarif WM,Ayyad SEN,et al.,2015.New cytotoxic isoprenoid derivatives from the Red Sea soft coral Sarcophyton glaucum.Nat Prod Res,29(1):24-30.https://doi.org/10.1080/14786419.2014.952637
Al-Lihaibi SS,Alarif WM,Abdel-Lateff A,et al.,2014.Three new cembranoid-type diterpenes from Red Sea soft coral Sarcophyton glaucum:isolation and antiproliferative activity against HepG2 cells.Eur J Med Chem,81:314-322.https://doi.org/10.1016/j.ejmech.2014.05.016
Bradford MM,1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal Biochem,72(1-2):248-254.https://doi.org/10.1016/0003-2697(76)90527-3
Chai TT,Law YC,Wong FC,et al.,2017.Enzyme-assisted discovery of antioxidant peptides from edible marine invertebrates:a review.Mar Drugs,15(2):42.https://doi.org/10.3390/md15020042
Chao CH,Li WL,Huang CY,et al.,2017.Isoprenoids from the soft coral Sarcophyton glaucum.Mar Drugs,15(7):202.https://doi.org/10.3390/md15070202
Chen LL,Song LY,Li TF,et al.,2013.A new antiproliferative and antioxidant peptide isolated from Arca subcrenata.Mar Drugs,11(6):1800-1814.https://doi.org/10.3390/md11061800
Chen YC,Chang HS,Wang CT,et al.,2009.Antioxidative activities of hydrolysates from duck egg white using enzymatic hydrolysis.Asian Austral J Anim Sci,22(11):1587-1593.https://doi.org/10.5713/ajas.2009.90119
Cordeiro,R.,van Ofwegen,L.,Williams,G.,2010.World List of Octocorallia.Sarcophyton glaucum(Quoy&Gaimard,1833).http://www.marinespecies.org/aphia.php?p=taxdetails&id=209621[Accessed on Sept.25,2017].
Daliri EBM,Oh DH,Lee BH,2017.Bioactive peptides.Foods,6(5):32.https://doi.org/10.3390/foods6050032
de Lumen BO,2005.Lunasin:a cancer-preventive soy peptide.Nutr Rev,63(1):16-21.https://doi.org/10.1111/j.1753-4887.2005.tb00106.x
Dennison SR,Whittaker M,Harris F,et al.,2006.Anticancerα-helical peptides and structure/function relationships underpinning their interactions with tumour cell membranes.Curr Protein Pept Sci,7(6):487-499.https://doi.org/10.2174/138920306779025611
Fabricius KE,Alderslade P,2001.Soft Corals and Sea Fans:AComprehensive Guide to the Tropical Shallow Water Genera of the Central-West Pacific,the Indian Ocean and the Red Sea.Australian Institute of Marine Science,Townsville,Australia,p.264.
Fahmy H,Zjawiony JK,Konoshima T,et al.,2006.Potent skin cancer chemopreventing activity of some novel semisynthetic cembranoids from marine sources.Mar Drugs,4(2):28-36.https://doi.org/10.3390/md402028
Fan XD,Bai L,Mao XL,et al.,2017.Novel peptides with anti-proliferation activity from the Porphyra haitanesis hydrolysate.Process Biochem,60:97-107.https://doi.org/10.1016/j.procbio.2017.05.018
Ferlay J,Soerjomataram I,Ervik M,et al.,2018.GLOBOCAN2018,Cancer tomorrow:estimated number of incident cases from 2018 to 2040,all cancers,both sexes,all ages.International Agency for Research on Cancer,Lyon,France.http://gco.iarc.fr/tomorrow/graphic-line?type=0&population=900&mode=population&sex=0&cancer=39&age_group=value&apc_male=0&apc_female=0[Accessed on Dec.20,2018].
Gabernet G,Müller AT,Hiss JA,et al.,2016.Membranolytic anticancer peptides.Med Chem Commun,7(12):2232-2245.https://doi.org/10.1039/C6MD00376A
He GQ,Xuan GD,Ruan H,et al.,2005.Optimization of angiotensin I-converting enzyme(ACE)inhibition by rice dregs hydrolysates using response surface methodology.JZhejiang Univ-Sci B(Biomed&Biotechnol),6(6):508-513.https://doi.org/10.1631/jzus.2005.B0508
Hegazy ME,El-Beih AA,Moustafa AY,et al.,2011.Cytotoxic cembranoids from the Red Sea soft coral Sarcophyton glaucum.Nat Prod Commun,6(12):1809-1812.
Hsu KC,Li-Chan ECY,Jao CL,2011.Antiproliferative activity of peptides prepared from enzymatic hydrolysates of tuna dark muscle on human breast cancer cell line MCF-7.Food Chem,126(2):617-622.https://doi.org/10.1016/j.foodchem.2010.11.066
Huang CY,Sung PJ,Uvarani C,et al.,2015.Glaucumolides Aand B,biscembranoids with new structural type from a cultured soft coral Sarcophyton glaucum.Sci Rep,5:15624.https://doi.org/10.1038/srep15624
Huang YB,Wang XF,Wang HY,et al.,2011.Studies on mechanism of action of anticancer peptides by modulation of hydrophobicity within a defined structural framework.Mol Cancer Ther,10(3):416-426.https://doi.org/10.1158/1535-7163.mct-10-0811
Hung CC,Yang YH,Kuo PF,et al.,2014.Protein hydrolysates from tuna cooking juice inhibit cell growth and induce apoptosis of human breast cancer cell line MCF-7.JFunct Foods,11:563-570.https://doi.org/10.1016/j.jff.2014.08.015
Jumeri,Kim SM,2011.Antioxidant and anticancer activities of enzymatic hydrolysates of solitary tunicate(Styela clava).Food Sci Biotechnol,20(4):1075-1085.https://doi.org/10.1007/s10068-011-0146-y
Kim EK,Kim YS,Hwang JW,et al.,2013.Purification and characterization of a novel anticancer peptide derived from Ruditapes philippinarum.Process Biochem,48(7):1086-1090.https://doi.org/10.1016/j.procbio.2013.05.004
Li Y,Yu JM,2015.Research progress in structure-activity relationship of bioactive peptides.J Med Food,18(2):147-156.https://doi.org/10.1089/jmf.2014.0028
Minkiewicz P,Dziuba J,Iwaniak A,et al.,2008.BIOPEPdatabase and other programs for processing bioactive peptide sequences.J AOAC Int,91(4):965-980.
Ngo DH,Vo TS,Ngo DN,et al.,2012.Biological activities and potential health benefits of bioactive peptides derived from marine organisms.Int J Biol Macromol,51(4):378-383.https://doi.org/10.1016/j.ijbiomac.2012.06.001
Nielsen PM,Petersen D,Dambmann C,2001.Improved method for determining food protein degree of hydrolysis J Food Sci,66(5):642-646.https://doi.org/10.1111/j.1365-2621.2001.tb04614.x
Nurdiani R,Vasiljevic T,Yeager T,et al.,2017.Bioactive peptides with radical scavenging and cancer cell cytotoxic activities derived from Flathead(Platycephalus fuscus)by-products.Eur Food Res Technol,243(4):627-637.https://doi.org/10.1007/s00217-016-2776-z
Pan X,Zhao YQ,Hu FY,et al.,2016.Anticancer activity of a hexapeptide from skate(Raja porosa)cartilage protein hydrolysate in HeLa cells.Mar Drugs,14(8):153.https://doi.org/10.3390/md14080153
Pangestuti R,Kim SK,2017.Bioactive peptide of marine origin for the prevention and treatment of non-communicable diseases.Mar Drugs,15(3):67.https://doi.org/10.3390/md15030067
Picot L,Bordenave S,Didelot S,et al.,2006.Antiproliferative activity of fish protein hydrolysates on human breast cancer cell lines.Process Biochem,41(5):1217-1222.https://doi.org/10.1016/j.procbio.2005.11.024
Qian ZJ,Je JY,Kim SK.,2007.Antihypertensive effect of angiotensin I converting enzyme-inhibitory peptide from hydrolysates of bigeye tuna dark muscle,Thunnus obesus.J Agric Food Chem,55(21):8398-8403.https://doi.org/10.1021/jf0710635
Quah Y,Ismail NIM,Ooi JLS,et al.,2017.Identification of novel cytotoxic peptide KENPVLSLVNGMF from marine sponge Xestospongia testudinaria,with characterization of stability in human serum.Int J Pept Res Ther,24(1):189-199.https://doi.org/10.1007/s10989-017-9604-6
Schmidt N,Mishra A,Lai GH,et al.,2010.Arginine-rich cell-penetrating peptides.FEBS Lett,584(9):1806-1813.https://doi.org/10.1016/j.febslet.2009.11.046
Shan YP,Huang JF,Tan JJ,et al.,2012.The study of single anticancer peptides interacting with HeLa cell membranes by single molecule force spectroscopy.Nanoscale,4(4):1283-1286.https://doi.org/10.1039/c2nr11541g
Simmons TL,Andrianasolo E,McPhail K,et al.,2005.Marine natural products as anticancer drugs.Mol Cancer Ther,4(2):333-342.
Song R,Wei RB,Luo HY,et al.,2014.Isolation and identification of an antiproliferative peptide derived from heated products of peptic hydrolysates of half-fin anchovy(Setipinna taty).J Funct Foods,10:104-111.https://doi.org/10.1016/j.jff.2014.06.010
Sutradhar KB,Amin ML,2014.Nanotechnology in cancer drug delivery and selective targeting.Int Sch Res Notices,2014:939378.https://doi.org/10.1155/2014/939378
Tada N,Horibe T,Haramoto M,et al.,2011.A single replacement of histidine to arginine in EGFR-lytic hybrid peptide demonstrates the improved anticancer activity.Biochem Biophys Res Commun,407(2):383-388.https://doi.org/10.1016/j.bbrc.2011.03.030
Wang B,Li L,Chi CF,et al.,2013.Purification and characterisation of a novel antioxidant peptide derived from blue mussel(Mytilus edulis)protein hydrolysate.Food Chem,138(2-3):1713-1719.https://doi.org/10.1016/j.foodchem.2012.12.002
Wang YK,He HL,Wang GF,et al.,2010.Oyster(Crassostrea gigas)hydrolysates produced on a plant scale have antitumor activity and immunostimulating effects in BALB/c mice.Mar Drugs,8(2):255-268.https://doi.org/10.3390/md8020255
Wang ZJ,Zhang XW,2016.Inhibitory effects of small molecular peptides from Spirulina(Arthrospira)platensis on cancer cell growth.Food Funct,7(2):781-788.https://doi.org/10.1039/C5FO01186H
You LJ,Zhao MM,Liu RH,et al.,2011.Antioxidant and antiproliferative activities of loach(Misgurnus anguillicaudatus)peptides prepared by papain digestion.J Agric Food Chem,59(14):7948-7953.https://doi.org/10.1021/jf2016368
Zheng LH,Lin XK,Wu N,et al.,2013.Targeting cellular apoptotic pathway with peptides from marine organisms.Biochim Biophys Acta,1836(1):42-48.https://doi.org/10.1016/j.bbcan.2013.02.006
Al-Lihaibi SS,Alarif WM,Abdel-Lateff A,et al.,2014.Three new cembranoid-type diterpenes from Red Sea soft coral Sarcophyton glaucum:isolation and antiproliferative activity against HepG2 cells.Eur J Med Chem,81:314-322.https://doi.org/10.1016/j.ejmech.2014.05.016
Bradford MM,1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal Biochem,72(1-2):248-254.https://doi.org/10.1016/0003-2697(76)90527-3
Chai TT,Law YC,Wong FC,et al.,2017.Enzyme-assisted discovery of antioxidant peptides from edible marine invertebrates:a review.Mar Drugs,15(2):42.https://doi.org/10.3390/md15020042
Chao CH,Li WL,Huang CY,et al.,2017.Isoprenoids from the soft coral Sarcophyton glaucum.Mar Drugs,15(7):202.https://doi.org/10.3390/md15070202
Chen LL,Song LY,Li TF,et al.,2013.A new antiproliferative and antioxidant peptide isolated from Arca subcrenata.Mar Drugs,11(6):1800-1814.https://doi.org/10.3390/md11061800
Chen YC,Chang HS,Wang CT,et al.,2009.Antioxidative activities of hydrolysates from duck egg white using enzymatic hydrolysis.Asian Austral J Anim Sci,22(11):1587-1593.https://doi.org/10.5713/ajas.2009.90119
Cordeiro,R.,van Ofwegen,L.,Williams,G.,2010.World List of Octocorallia.Sarcophyton glaucum(Quoy&Gaimard,1833).http://www.marinespecies.org/aphia.php?p=taxdetails&id=209621[Accessed on Sept.25,2017].
Daliri EBM,Oh DH,Lee BH,2017.Bioactive peptides.Foods,6(5):32.https://doi.org/10.3390/foods6050032
de Lumen BO,2005.Lunasin:a cancer-preventive soy peptide.Nutr Rev,63(1):16-21.https://doi.org/10.1111/j.1753-4887.2005.tb00106.x
Dennison SR,Whittaker M,Harris F,et al.,2006.Anticancerα-helical peptides and structure/function relationships underpinning their interactions with tumour cell membranes.Curr Protein Pept Sci,7(6):487-499.https://doi.org/10.2174/138920306779025611
Fabricius KE,Alderslade P,2001.Soft Corals and Sea Fans:AComprehensive Guide to the Tropical Shallow Water Genera of the Central-West Pacific,the Indian Ocean and the Red Sea.Australian Institute of Marine Science,Townsville,Australia,p.264.
Fahmy H,Zjawiony JK,Konoshima T,et al.,2006.Potent skin cancer chemopreventing activity of some novel semisynthetic cembranoids from marine sources.Mar Drugs,4(2):28-36.https://doi.org/10.3390/md402028
Fan XD,Bai L,Mao XL,et al.,2017.Novel peptides with anti-proliferation activity from the Porphyra haitanesis hydrolysate.Process Biochem,60:97-107.https://doi.org/10.1016/j.procbio.2017.05.018
Ferlay J,Soerjomataram I,Ervik M,et al.,2018.GLOBOCAN2018,Cancer tomorrow:estimated number of incident cases from 2018 to 2040,all cancers,both sexes,all ages.International Agency for Research on Cancer,Lyon,France.http://gco.iarc.fr/tomorrow/graphic-line?type=0&population=900&mode=population&sex=0&cancer=39&age_group=value&apc_male=0&apc_female=0[Accessed on Dec.20,2018].
Gabernet G,Müller AT,Hiss JA,et al.,2016.Membranolytic anticancer peptides.Med Chem Commun,7(12):2232-2245.https://doi.org/10.1039/C6MD00376A
He GQ,Xuan GD,Ruan H,et al.,2005.Optimization of angiotensin I-converting enzyme(ACE)inhibition by rice dregs hydrolysates using response surface methodology.JZhejiang Univ-Sci B(Biomed&Biotechnol),6(6):508-513.https://doi.org/10.1631/jzus.2005.B0508
Hegazy ME,El-Beih AA,Moustafa AY,et al.,2011.Cytotoxic cembranoids from the Red Sea soft coral Sarcophyton glaucum.Nat Prod Commun,6(12):1809-1812.
Hsu KC,Li-Chan ECY,Jao CL,2011.Antiproliferative activity of peptides prepared from enzymatic hydrolysates of tuna dark muscle on human breast cancer cell line MCF-7.Food Chem,126(2):617-622.https://doi.org/10.1016/j.foodchem.2010.11.066
Huang CY,Sung PJ,Uvarani C,et al.,2015.Glaucumolides Aand B,biscembranoids with new structural type from a cultured soft coral Sarcophyton glaucum.Sci Rep,5:15624.https://doi.org/10.1038/srep15624
Huang YB,Wang XF,Wang HY,et al.,2011.Studies on mechanism of action of anticancer peptides by modulation of hydrophobicity within a defined structural framework.Mol Cancer Ther,10(3):416-426.https://doi.org/10.1158/1535-7163.mct-10-0811
Hung CC,Yang YH,Kuo PF,et al.,2014.Protein hydrolysates from tuna cooking juice inhibit cell growth and induce apoptosis of human breast cancer cell line MCF-7.JFunct Foods,11:563-570.https://doi.org/10.1016/j.jff.2014.08.015
Jumeri,Kim SM,2011.Antioxidant and anticancer activities of enzymatic hydrolysates of solitary tunicate(Styela clava).Food Sci Biotechnol,20(4):1075-1085.https://doi.org/10.1007/s10068-011-0146-y
Kim EK,Kim YS,Hwang JW,et al.,2013.Purification and characterization of a novel anticancer peptide derived from Ruditapes philippinarum.Process Biochem,48(7):1086-1090.https://doi.org/10.1016/j.procbio.2013.05.004
Li Y,Yu JM,2015.Research progress in structure-activity relationship of bioactive peptides.J Med Food,18(2):147-156.https://doi.org/10.1089/jmf.2014.0028
Minkiewicz P,Dziuba J,Iwaniak A,et al.,2008.BIOPEPdatabase and other programs for processing bioactive peptide sequences.J AOAC Int,91(4):965-980.
Ngo DH,Vo TS,Ngo DN,et al.,2012.Biological activities and potential health benefits of bioactive peptides derived from marine organisms.Int J Biol Macromol,51(4):378-383.https://doi.org/10.1016/j.ijbiomac.2012.06.001
Nielsen PM,Petersen D,Dambmann C,2001.Improved method for determining food protein degree of hydrolysis J Food Sci,66(5):642-646.https://doi.org/10.1111/j.1365-2621.2001.tb04614.x
Nurdiani R,Vasiljevic T,Yeager T,et al.,2017.Bioactive peptides with radical scavenging and cancer cell cytotoxic activities derived from Flathead(Platycephalus fuscus)by-products.Eur Food Res Technol,243(4):627-637.https://doi.org/10.1007/s00217-016-2776-z
Pan X,Zhao YQ,Hu FY,et al.,2016.Anticancer activity of a hexapeptide from skate(Raja porosa)cartilage protein hydrolysate in HeLa cells.Mar Drugs,14(8):153.https://doi.org/10.3390/md14080153
Pangestuti R,Kim SK,2017.Bioactive peptide of marine origin for the prevention and treatment of non-communicable diseases.Mar Drugs,15(3):67.https://doi.org/10.3390/md15030067
Picot L,Bordenave S,Didelot S,et al.,2006.Antiproliferative activity of fish protein hydrolysates on human breast cancer cell lines.Process Biochem,41(5):1217-1222.https://doi.org/10.1016/j.procbio.2005.11.024
Qian ZJ,Je JY,Kim SK.,2007.Antihypertensive effect of angiotensin I converting enzyme-inhibitory peptide from hydrolysates of bigeye tuna dark muscle,Thunnus obesus.J Agric Food Chem,55(21):8398-8403.https://doi.org/10.1021/jf0710635
Quah Y,Ismail NIM,Ooi JLS,et al.,2017.Identification of novel cytotoxic peptide KENPVLSLVNGMF from marine sponge Xestospongia testudinaria,with characterization of stability in human serum.Int J Pept Res Ther,24(1):189-199.https://doi.org/10.1007/s10989-017-9604-6
Schmidt N,Mishra A,Lai GH,et al.,2010.Arginine-rich cell-penetrating peptides.FEBS Lett,584(9):1806-1813.https://doi.org/10.1016/j.febslet.2009.11.046
Shan YP,Huang JF,Tan JJ,et al.,2012.The study of single anticancer peptides interacting with HeLa cell membranes by single molecule force spectroscopy.Nanoscale,4(4):1283-1286.https://doi.org/10.1039/c2nr11541g
Simmons TL,Andrianasolo E,McPhail K,et al.,2005.Marine natural products as anticancer drugs.Mol Cancer Ther,4(2):333-342.
Song R,Wei RB,Luo HY,et al.,2014.Isolation and identification of an antiproliferative peptide derived from heated products of peptic hydrolysates of half-fin anchovy(Setipinna taty).J Funct Foods,10:104-111.https://doi.org/10.1016/j.jff.2014.06.010
Sutradhar KB,Amin ML,2014.Nanotechnology in cancer drug delivery and selective targeting.Int Sch Res Notices,2014:939378.https://doi.org/10.1155/2014/939378
Tada N,Horibe T,Haramoto M,et al.,2011.A single replacement of histidine to arginine in EGFR-lytic hybrid peptide demonstrates the improved anticancer activity.Biochem Biophys Res Commun,407(2):383-388.https://doi.org/10.1016/j.bbrc.2011.03.030
Wang B,Li L,Chi CF,et al.,2013.Purification and characterisation of a novel antioxidant peptide derived from blue mussel(Mytilus edulis)protein hydrolysate.Food Chem,138(2-3):1713-1719.https://doi.org/10.1016/j.foodchem.2012.12.002
Wang YK,He HL,Wang GF,et al.,2010.Oyster(Crassostrea gigas)hydrolysates produced on a plant scale have antitumor activity and immunostimulating effects in BALB/c mice.Mar Drugs,8(2):255-268.https://doi.org/10.3390/md8020255
Wang ZJ,Zhang XW,2016.Inhibitory effects of small molecular peptides from Spirulina(Arthrospira)platensis on cancer cell growth.Food Funct,7(2):781-788.https://doi.org/10.1039/C5FO01186H
You LJ,Zhao MM,Liu RH,et al.,2011.Antioxidant and antiproliferative activities of loach(Misgurnus anguillicaudatus)peptides prepared by papain digestion.J Agric Food Chem,59(14):7948-7953.https://doi.org/10.1021/jf2016368
Zheng LH,Lin XK,Wu N,et al.,2013.Targeting cellular apoptotic pathway with peptides from marine organisms.Biochim Biophys Acta,1836(1):42-48.https://doi.org/10.1016/j.bbcan.2013.02.006