套膜海葵总蛋白的提取和酶解条件优化及其杀虫活性  

Extraction and Enzymolysis Conditions Optimization of Aiptasia pallida Total Protein and Its Insecticidal Activity

作　　者：袁琳 符金星 刘雪菲丹 高炳淼[1] YUAN Lin;FU Jinxing;LIU Xuefeidan;GAO Bingmiao(School of Pharmacy/Key Laboratory of Tropical Translational Medicine of Ministry of Education,Hainan Medical University,Haikou,Hainan 571199,China)

机构地区：[1]海南医学院药学院/热带转化医学教育部重点实验室,海南海口571199

出　　处：《贵州农业科学》2021年第3期50-55,共6页Guizhou Agricultural Sciences

基　　金：国家自然科学基金项目“多组学整合技术发掘南海海葵多肽基因资源及其杀虫作用机制研究”(82060686);海南省重点研发计划项目“南海海葵毒素的抗虫活性筛选及其生物杀虫剂的应用研究”(ZDYF2018138);海南省自然科学基金项目“海葵多肽的化学合成及其杀虫作用机制研究”(820RC636)。

摘　　要：【目的】明确套膜海葵总蛋白的提取工艺、最优酶解条件及其杀虫活性,为拓展和开发海葵酶解多肽的应用提供理论依据。【方法】采用裂解液法提取海葵总蛋白,结合单因素试验对其酶解条件进行优化;同时采用昆虫注射法研究酶解海葵多肽的杀虫活性。【结果】从套膜海葵中提取海葵总蛋白的分子量主要为31.0~43.0kDa,20g套膜海葵可提取海葵总蛋白160mg,蛋白浓度为0.5mg/mL;海葵总蛋白的最优酶解条件为pH 8、碱性蛋白酶(最佳水解酶)4000U/g、温度60℃、酶解6h,在此条件下酶解得率为9.145%;昆虫注射海葵酶解多肽1.0μg/mg时死亡率达90%。【结论】获得海葵总蛋白酶解的最佳蛋白酶及其最优酶解条件,成功制备具有杀虫活性的海葵酶解多肽,为深入开发海葵酶解多肽的杀虫活性奠定了基础。【Objective】The extraction technology,optimum enzymolysis condition and insecticidal activity of Aiptasia pallida total protein is clarified to provide the theoretical basis for expanding and exploiting the application of sea anemone enzymolysis polypeptide.【Method】Sea anemone total protein is extracted by lysis solution method,its enzymolysis conditions are optimized combined with a single factor experiment.The insecticidal activity of enzymolysis sea anemone polypeptide is studied by using insect injection method at the same time.【Result】The molecular weight of sea anemone total protein extracted fromA.pallida is 31.0—43.0 kDa,and 160 mg sea anemone total protein with 0.5 mg/mL protein concentration is extracted from 20 g A.pallida.The optimum enzymolysis conditions of sea anemone total protein include pH 8,4000 U/g alkaline protease(optimum hydrolase),and enzymolysis 6 hat 60℃.The enzymolysis yield under the optimum conditions is 9.145%.The morality is up to 90%when insects inject1.0μg/mg sea anemone enzymolysis polypeptide.【Conclusion】The optimum protease and optimum enzymolysis polypeptide with insecticidal activity is prepared successfully,which lay the foundation for deeply exploitation of insecticidal activity in sea anemone enzymolysis polypeptide.

关 键 词：海葵 海葵毒素 多肽 酶解 杀虫活性 

分 类 号：S482.39[农业科学—农药学]