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Preparation of Sea Anemone Peptide Toxin Ap-TxI and Its Insecticidal Activity

查看全文 作　　者：Yongyi [1]XU;Yanling [1]LIAO;Qiqi [1]GUO;Ming [1]LI;Jinxing [1]FU;Bingmiao [1]GAO 高影响力作者 机构地区：[1]Key Laboratory of Tropical Translational Medicine of Ministry of Education,College of Pharmacy,Hainan Medical University,Haikou 571199,China高影响力机构 出　　处：《Agricultural Biotechnology》索引2022年第11卷第5期,共4页高影响力期刊 基　　金：Supported by Natural Science Foundation of Hainan Province (820RC636);Undergraduate Innovation and Enterpreneurship Training Program of Hainan Province (X202011810003);Special Fund for Academician Innovation Platform in Hainan Province (YSPTZX202132)。 摘　　要：[Objectives]This study was conducted to synthesize sea anemone peptide toxin Ap-TxI and investigate its insecticidal activity. [Methods] The sea anemone linear peptide toxin Ap-TxI was synthesized by the solid-phase peptide synthesis(SPPS), and six cysteines were oxidized to form three disulfide bonds by a three-step directional oxidation method. Then, purification by high performance liquid chromatography(HPLC) and mass spectrometry identification were performed. Finally, the insect cytotoxicity and insecticidal activity of Ap-TxI were studied by the MTT method and insect injection method. [Results] The oxidized peptide Ap-TxI with three disulfide bonds in natural configuration was successfully synthesized by the SPPS method, and its purity was >90% by HPLC analysis. The results of the MTT method showed that Ap-TxI was active on the growth of insect cells sf9, with a half effective dose of 0.2 nM;and the results of the mealworm injection test showed that the polypeptide Ap-TxI had high insecticidal activity with a median lethal dose of 11.7 nM. [Conclusions] The sea anemone peptide toxin Ap-TxI with high insecticidal effect was obtained, laying a foundation for the development of new, efficient and safe biological insecticides. 关 键 词：Sea anemone peptide toxin Solid-phase synthesis Oxidative folding Insecticidal activity
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