酶解条浒苔蛋白制备抗肺癌活性多肽
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Marine Algae，Quanzhou 362000）
Abstract: In order to exploit abundant proteins in the green algae Enteromorpha clathrata，protein from E. clathrata was hydrolyzed by the papain enzyme，the optimal hydrolysis process conditions were solid/liquid ratio of 1∶25，enzyme dose of 1 250 U/g protein，hydrolysis temperature of 45.7℃，hydrolysis pH of 7.25 and hydrolysis duration of 120 min. Under these conditions，polypeptides of different molecular weight range were obtained by ultrafiltration，and their anti-lung cancer effects were evaluated preliminarily at the cellular level of HUVEC （human umbilical vein endothelial cell），A549，H446 and H460. According to the results，the oxidation resistance，proliferation and tubule formation of HUVEC treated with 2-6 kD polypeptide from E. clathrata were inhibited，and the proliferation，mobility，cell cycle of all 3 tested lung cancer cells H446，H460 and A549 were inhibited at varied level，and the cells apoptosis were induced. The 2-6 kD proteins demonstrated significantly better inhibitory effect on H446 and H460 than that on A549. In conclusion，this study proves the feasibility of gaining natural anti-tumor small-molecule peptides by enzymolysis protein from E. clathrata，and provides a novel way to explore the high value utilization of protein resources from E. clathrata.
Key words: Enteromorpha clathrata ；enzymatic hydrolysis ；peptide ；lung cancer
DUAN Xun-wei1，4 XIAO Gui-qing2 WANG Li-xing3 WU Wen-lin1，4 DAI Cong-jie1，4 DONG Le1，4
（1. Quanzhou Normal University，Quanzhou 362000 ；2.Yinfeng Stem Cell Engineering Co.，Quanzhou 362000 ；3.Second Affiliated Hospital，Fujian Medical University，Quanzhou 362000 ；4.Fujian Province Key Laboratory for the Development of Bioactive Material from
摘 要 ： 为了开发利用绿潮藻类条浒苔中含量丰富的蛋白质，采用木瓜蛋白酶酶解条浒苔蛋白，最佳酶解条件为 ：料液比 1∶25、加酶量 1 250 U/g pro、温度 45.7℃、pH 7.2、震荡酶解时间 120 min。在该条件下的酶解多肽经超滤分离获得不同分子量区 间的多肽，并通过 HUVEC、A549、H446、H460 等细胞水平初步评价酶解条浒苔制备多肽抗肺癌的细胞生物学效果。结果表明， 经上述条件获得的 2-6 kD 条浒苔多肽处理的 HUVEC 在抗氧化和小管形成等方面均受到抑制，其处理的 NCI-H460、NCI-H446 和 A549 的增殖、细胞周期、迁移也受到不同程度抑制，且能够促进细胞凋亡 ；2-6 kD 浒苔多肽对 H446、H460 作用效果显著优于对 A549 的作用。这一细胞水平研究，不仅证实了酶解条浒苔获取新型天然抗肿瘤小肽的可行性，也为条浒苔蛋白质资源高值化利用 探索提供了新的途径。
·技术与方法·
生物技术报
BIOTECHNOLOGY BULLETIN
2019, 35(8):246-252
酶解条浒苔蛋白制备抗肺癌活性多肽
段训威1，4 肖桂清2 王礼兴3 吴文林1，4 戴聪杰1，4 董乐1，4
（1. 泉州师范学院，泉州 362000 ；2. 福建省银丰干细胞工程有限公司，泉州 362000 ；3. 福建医科大学附属第二医院，泉州 362000 ；4. 福 建省海洋藻类活性物质制备与功能开发重点实验室，泉州 362000）
关键词 ： 条浒苔 ；酶解 ；多肽 ；肺癌 DOI ：10.13560/ki.biotech.bull.1985.2019-0338
Preparation of Anti-lung Cancer Peptides by Enzymatic Hydrolysis of
Protein from Enteromorpha clathrata
Abstract: In order to exploit abundant proteins in the green algae Enteromorpha clathrata，protein from E. clathrata was hydrolyzed by the papain enzyme，the optimal hydrolysis process conditions were solid/liquid ratio of 1∶25，enzyme dose of 1 250 U/g protein，hydrolysis temperature of 45.7℃，hydrolysis pH of 7.25 and hydrolysis duration of 120 min. Under these conditions，polypeptides of different molecular weight range were obtained by ultrafiltration，and their anti-lung cancer effects were evaluated preliminarily at the cellular level of HUVEC （human umbilical vein endothelial cell），A549，H446 and H460. According to the results，the oxidation resistance，proliferation and tubule formation of HUVEC treated with 2-6 kD polypeptide from E. clathrata were inhibited，and the proliferation，mobility，cell cycle of all 3 tested lung cancer cells H446，H460 and A549 were inhibited at varied level，and the cells apoptosis were induced. The 2-6 kD proteins demonstrated significantly better inhibitory effect on H446 and H460 than that on A549. In conclusion，this study proves the feasibility of gaining natural anti-tumor small-molecule peptides by enzymolysis protein from E. clathrata，and provides a novel way to explore the high value utilization of protein resources from E. clathrata.
Key words: Enteromorpha clathrata ；enzymatic hydrolysis ；peptide ；lung cancer
DUAN Xun-wei1，4 XIAO Gui-qing2 WANG Li-xing3 WU Wen-lin1，4 DAI Cong-jie1，4 DONG Le1，4
（1. Quanzhou Normal University，Quanzhou 362000 ；2.Yinfeng Stem Cell Engineering Co.，Quanzhou 362000 ；3.Second Affiliated Hospital，Fujian Medical University，Quanzhou 362000 ；4.Fujian Province Key Laboratory for the Development of Bioactive Material from
摘 要 ： 为了开发利用绿潮藻类条浒苔中含量丰富的蛋白质，采用木瓜蛋白酶酶解条浒苔蛋白，最佳酶解条件为 ：料液比 1∶25、加酶量 1 250 U/g pro、温度 45.7℃、pH 7.2、震荡酶解时间 120 min。在该条件下的酶解多肽经超滤分离获得不同分子量区 间的多肽，并通过 HUVEC、A549、H446、H460 等细胞水平初步评价酶解条浒苔制备多肽抗肺癌的细胞生物学效果。结果表明， 经上述条件获得的 2-6 kD 条浒苔多肽处理的 HUVEC 在抗氧化和小管形成等方面均受到抑制，其处理的 NCI-H460、NCI-H446 和 A549 的增殖、细胞周期、迁移也受到不同程度抑制，且能够促进细胞凋亡 ；2-6 kD 浒苔多肽对 H446、H460 作用效果显著优于对 A549 的作用。这一细胞水平研究，不仅证实了酶解条浒苔获取新型天然抗肿瘤小肽的可行性，也为条浒苔蛋白质资源高值化利用 探索提供了新的途径。
·技术与方法·
生物技术报
BIOTECHNOLOGY BULLETIN
2019, 35(8):246-252
酶解条浒苔蛋白制备抗肺癌活性多肽
段训威1，4 肖桂清2 王礼兴3 吴文林1，4 戴聪杰1，4 董乐1，4
（1. 泉州师范学院，泉州 362000 ；2. 福建省银丰干细胞工程有限公司，泉州 362000 ；3. 福建医科大学附属第二医院，泉州 362000 ；4. 福 建省海洋藻类活性物质制备与功能开发重点实验室，泉州 362000）
关键词 ： 条浒苔 ；酶解 ；多肽 ；肺癌 DOI ：10.13560/ki.biotech.bull.1985.2019-0338
Preparation of Anti-lung Cancer Peptides by Enzymatic Hydrolysis of
Protein from Enteromorpha clathrata