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1、bAntifungalandAnticancerActivitiesofaProteinfromtheMushroomCordycepsmilitarisThemushroomCordycepsmilitarishasbeenusedforalongtimeineasternAsiaasanutraceuticalandintraditionalChinesemedicineasatreatmentforcancerpatients.Inthepresentstudy,acytotoxicantifungalproteasewaspurifiedfromth
2、edriedfruitingbodiesofC.militarisusinganion-exchangechromatographyonaDEAE-Sepharosecolumn.Electrophoreticanalysesindicatedthatthisprotein,designatedC.militarisprotein(CMP),hasamolecularmassof12kDaandapIof5.1.Theoptimumconditionsforproteaseactivitywereatemperatureof37oCandpHof7.0∼9.
3、0.Theenzymeactivitywasspecificallyinhibitedbytheserineproteaseinhibitorphenylmethylsulfonylfluoride.AminoacidcompositionofintactCMPandaminoacidsequencesofthreemajorpeptidesfromatrypticdigestofCMPweredetermined.CMPexertedstrongantifungaleffectagainstthegrowthofthefungusFusariumoxysp
4、orum,andexhibitedcytotoxicityagainsthumanbreastandbladdercancercells.TheseresultsindicatethatC.militarisrepresentsasourceofanovelproteinthatmightbeappliedindiversebiologicalandmedicinalapplications.菌类蛹虫草在东亚作为保健品和中国传统医学中作为癌症患者的治疗药物已经有很长一段时间的历史了。在本研究中，通过在DEAE-琼脂糖凝胶柱上使用阴离子交换层析法，研究者从干的
5、蛹虫草子中分离纯化出了一种具有细胞毒性的抗真菌蛋白酶。电泳分析表明，这种被称为蛹虫草蛋白（CMP）的蛋白质有12kDa的分子量，其等电点为5.1。它的蛋白酶活性的最佳条件为温度37℃，pH值7.0〜9.0。这种酶的活性尤其受到丝氨酸蛋白酶抑制剂--苯甲基磺酰氟的抑制。完整的蛹虫草蛋白（CMP）的氨基酸组成和三个主要的蛹虫草蛋白（CMP）的胰蛋白酶消化肽的氨基酸序列已经被测定。CMP对菌枯萎病菌的生长产生很强的抑菌作用，并表现出对人类乳腺癌和膀胱癌细胞的胞毒作用。这些结果表明，蛹虫草代表了一种新的蛋白质可能被应用在不同的生物和药物领域.KeyWords:Mus
6、hroom,Cordycepsmilitaris,Protease,Antifungalactivity,bbCytotoxicity关键词：菌类，蛹虫草，蛋白酶，抗真菌活性，细胞毒性INTRODUCTIONMushroomscontainnumerouscompoundsthatexhibitinterestingbiologicalactivities,includingantibacterial,antifungal,antiviral,antitumor,antiproliferative,immunomodulatory,lectin-like,p
7、rotease,andnucleaseactivities(Ng2004;Lindequistetal,2005).ThemushroomCordycepsmilitarisisaspeciesofentomogenousfungusthatisparasiticoninsects,attackingtheundergroundpupaeandlarvaeoflepidopteraninsects.Thesefungiproduceinhibitorsofthephenoloxidase-activatingsystemsofinsects,andthent
8、heysecretevariousenzymesth
