Full Text:  <1780>

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CLC number: Q516

On-line Access: 2019-01-07

Received: 2017-12-01

Revision Accepted: 2018-04-19

Crosschecked: 2018-12-05

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Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Tsun-Thai Chai

https://orcid.org/0000-0003-3716-1599

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Journal of Zhejiang University SCIENCE B

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


Author(s):  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

Affiliation(s):  Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman, 31900 Kampar, Malaysia; more

Corresponding email(s):  chaitt@utar.edu.my

Key Words:  Anticancer therapy; Bioactive peptide; Cytotoxicity; HeLa cells; Sarcophyton glaucum; Soft coral


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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. Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum[J]. Journal of Zhejiang University Science B, 2019, 20(7): 59-70.

@article{title="Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum",
author="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",
journal="Journal of Zhejiang University Science B",
volume="20",
number="1",
pages="59-70",
year="2019",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1700586"
}

%0 Journal Article
%T Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum
%A Yixian Quah
%A Nor Ismaliza Mohd Ismail
%A Jillian Lean Sim Ooi
%A Yang Amri Affendi
%A Fazilah Abd Manan
%A Lai-Kuan Teh
%A Fai-Chu Wong
%A Tsun-Thai Chai
%J Journal of Zhejiang University SCIENCE B
%V 20
%N 1
%P 59-70
%@ 1673-1581
%D 2019
%I Zhejiang University Press & Springer

TY - JOUR
T1 - Purification and identification of novel cytotoxic oligopeptides from soft coral Sarcophyton glaucum
A1 - Yixian Quah
A1 - Nor Ismaliza Mohd Ismail
A1 - Jillian Lean Sim Ooi
A1 - Yang Amri Affendi
A1 - Fazilah Abd Manan
A1 - Lai-Kuan Teh
A1 - Fai-Chu Wong
A1 - Tsun-Thai Chai
J0 - Journal of Zhejiang University Science B
VL - 20
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PB - Zhejiang University Press & Springer
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Abstract: 
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.

软珊瑚Sarcophyton glaucum中新型细胞毒性寡肽的纯化与鉴定

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HeLa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.

关键词组:抗癌治疗;活性肽;细胞毒性;HeLa细胞;Sarcophyton glaucum;软珊瑚

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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[47]Fig. S1 A representative SPE elution prolife of GF3

[48]Fig. S2 Peptide content of SPE fractions

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