中国毛虾流感病毒神经氨酸酶抑制活性肽的研究

以中国毛虾为原料, 以抑制流感病毒神经氨酸酶(NA)活性为初筛指标, 通过控制酶切位点制备了具有抑制NA活性的酶解液. 利用凝胶层析和高效液相色谱等技术分离纯化出高活性的抑制肽, 其IC₅₀ 值为96.1 μmol/L.经串联质谱测定该抑制肽序列为EISYIHAEAYRRGELK, 紫外光谱分析结果证明该抑制肽能与NA结合.基于反向对接, 应用SYBYL软件模拟抑制肽与NA活性区域结合, 确定了抑制肽与NA的结合位点. 细胞毒性实验测得该抑制肽对细胞的最大无毒浓度(TC₀)为1.26 mg/mL.在红细胞凝集实验中, 随着抑制肽浓度增大, 病毒的凝集价显著降低, 证明抑制肽的抗病毒作用具有多靶点.

Active Peptide from Acetes Chinensis with Inhibitory Activity on Neuraminidase of Influenza Virus

Low-valued Acetes Chinensis was chosen to prepare the enzymatic hydrolysate with neuraminidase(NA) inhibitory activity by controlled hydrolysis. A novel NA inhibitory peptide was purified by gel chromatography and reversed phase high performance liquid chromatography. The IC₅₀ value of the peptide was only 96.1 μmol/L and it could also significantly inhibit influenza A virus replication in MDCK cells. The amino acid sequence of this peptide was EISYIHAEAYRRGELK which identified by LC-MS/MS and its binding with NA protein was proved by UV spectrum analysis. The accurate sites of NA protein that the peptide bind to were determined using the computer assisted software SYBYL. The largest non-toxic concentration(TC₀) was 1.26 mg/mL which assayed by cell cytotoxicity experiments. The hemagglutination essay showed that this pepetide could obviously decrease the agglutination titer influenza virus in a dose-dependent manner which suggested that the antiviral activity of this peptide might be attributed to its action on multiple targets.