食品科学2017，Vol.38Issue(12)：27-35,9.DOI:10.7506/spkx1002-6630-201712005
双齿围沙蚕多肽的制备及其抗肺癌A549细胞活性
Anticancer Activity of a Novel Peptide Derived from Hydrolysates of Perinereies aibuhitensis against Lung Cancer A549 Cells
摘要
Abstract
The key steps for preparing antitumor peptide from enzymatic hydrolysates of Perinereies aibuhitensis were investigated in the present work.The most suitable enzyme preparation was selected and the optimal hydrolysis conditions allowing high inhibition of cancer cell proliferation were determined using one-factor-at-a-time method and orthogonal array design.A fraction containing peptides with molecular weights of 1-3 ku was obtained by ultrafilation of hydrolysates,and it was further purified by sequential ion exchange chromatography,gel filtration chromatography and preparative chromatography.Then,lung cancer A549 cells were used to test the anticaner effect of the purified peptide PAP by methylthiazolyldiphenyl-tetrazolium bromide (MTr) assay.The morphological changes were observed through an inverted microscope,dual acridine orange/ethidium bromide (AO/EB) fluorescent staining and Hoechst fluorescent staining.The results showed that alcalase was found to be the optimal enzyme for the production of anticancer peptide and that the optimal hydrolysis conditions were determined as follows:temperature,50 ℃;pH value,11;solid-to-liquid ratio,1∶1;hydrolysis time,6 h;and enzyme dosage,300 U/g.The peptide PAP was identified as Ile-Glu-Pro-Gly-Thr-Val-Gly-Met-Met-Phe.Moreover,our results demonstrated that PAP suppressed the proliferation of A549 cells in a time-and dose-dependent manner with morphological features of apoptosis being observed.Therefore,our findings suggest that PAP could inhibit the proliferation of lung cancer A549 cells and induced apoptosis.关键词
双齿围沙蚕/多肽/抗肿瘤/A549细胞/细胞凋亡Key words
Perinereies aibuhitensis/polypeptide/anti-cancer/A549 cell lines/cell apoptosis分类
医药卫生引用本文复制引用
贾盈露,丁国芳,杨最素,余方苗,郑媛媛,吴宗泽,陈锐..双齿围沙蚕多肽的制备及其抗肺癌A549细胞活性[J].食品科学,2017,38(12):27-35,9.基金项目
国家自然科学基金青年科学基金项目(81001393) （81001393）
2015年度国家星火计划项目(2015GA700044) （2015GA700044）
国家海洋重大计划项目(2015862) （2015862）
浙江省科技厅重大专项(2013C03036) （2013C03036）
浙江省自然科学基金项目(LS15H30001) （LS15H30001）
浙江省自然科学基金青年资金项目(LQ16H300001) （LQ16H300001）
舟山市科技计划项目(2015C31012) （2015C31012）
