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Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2019年第1期20卷 59-70页

作者： yixian quah Nor Ismaliza MOHD ISMAIL Jillian Lean Sim OOI Yang Amri AFFENDI Fazilah ABD MANAN Lai-Kuan TEH Fai-Chu WONG Tsun-Thai CHAI Department of Chemical Science Faculty of Science Universiti Tunku Abdul Rahman Department of Biological Science Faculty of Science Universiti Tunku Abdul Rahman Centre for Biodiversity Research Universiti Tunku Abdul Rahman Department of Geography Faculty of Arts and Social Sciences University of Malaya Institute of Biological Sciences Faculty of Science University of Malaya Institute of Ocean and Earth Sciences University of Malaya Department of Biosciences Faculty of Science Universiti Teknologi Malaysia Department of Biomedical Science Faculty of Science Universiti Tunku Abdul Rahman

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.

关键词： Anticancer therapy Bioactive peptide Cytotoxicity HeLa cells Sarcophyton glaucum Soft coral

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