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棉铃虫幼虫中肠主要蛋白酶活性的鉴定
引用本文:王琛柱,钦俊德.棉铃虫幼虫中肠主要蛋白酶活性的鉴定[J].昆虫学报,1996,39(1):7-14.
作者姓名:王琛柱  钦俊德
作者单位:中国科学院动物研究所
摘    要:根据棉铃虫Helicoverpa armigera(Hubner)中肠酶液对蛋白酶专性底物在不同pH下的水解作用,棉铃虫中肠的3种丝氨酸蛋白酶得到鉴定。它们是:强碱性类胰蛋白酶,水 解a-N-苯甲酰-DL-精氨酸-p-硝基苯胺的最适pH在10.50以上;弱碱性类胰蛋白酶,水解p-甲苯磺酰-L-精氨酸甲酯的最适pH为8.50~9.00;类胰凝乳蛋白酶, 水解N一苯甲酰-L-酪氨酸乙酯的最适pH亦为8.50-9.00。中肠总蛋白酶活性用偶 氮酪蛋白测定,最适pH亦在10.50以上。Ca2+对昆虫蛋白酶无影响,Mg2+仅对弱碱性类胰蛋白酶有激活作用。对苯甲基磺酰氟和甲基磺酰-L-赖氨酸氯甲基酮对弱碱性类胰蛋白酶的抑制作用较强,而对强碱性类胰蛋白酶的抑制作用较弱。甲基磺酰-L苯丙氨酸氯甲基酮除能抑制类胰凝乳蛋白酶外,还能激活弱碱性类胰蛋白酶。对牛胰蛋白酶有强抑制作用的卵粘蛋白抑制剂对昆虫蛋白酶却无抑制作用。大豆胰蛋白酶抑制剂对该虫的3种丝氨酸蛋白酶均有强的抑制作用。

关 键 词:棉铃虫  蛋白酶  激活剂  抑制剂  

PARTIAL CHARACTERIZATION OF PROTEASE ACTIVITY IN THE MIDGUT OF HELICOVERPA ARMIGERA LARVAE
Wang Chenzhu,Qin Junde.PARTIAL CHARACTERIZATION OF PROTEASE ACTIVITY IN THE MIDGUT OF HELICOVERPA ARMIGERA LARVAE[J].Acta Entomologica Sinica,1996,39(1):7-14.
Authors:Wang Chenzhu  Qin Junde
Abstract:The activities of three serine proteases in the midgut of Helicoperpaarmigera larvae were partially characterized.The enzymes were an active alkalinetrypsin-like enzyme with maximal hydrolysis of benzoyl arginine p-nitroanilideat pH 10.50 or higher,a weak alkaline trypsin-like enzyme with maximalhydrolysis of tosyl-L-arginine methyl ester at pH 8.50 -9. 00,and achymotrypsin-like enzyme with maximal hydrolysis of benzoyl-L-tyrosine ethylester at pH 8. 50-9. 00.Total proteolysis,measured by using azocasein, hadamaximal activity at pH 10. 50 or higher.Ca2+ had no activation effect on larvalproteases,but Mg2+ activated the weak alkaline trypsin-like enzyme.The inhibition with phenyl methyl sulfonyl fluoride and tosyl-L-lysine chloromethyl ketoneto the weak alkaline trypsin-like enzyme activity was greater than that to the active alkaline trypsin-like enzyme activity.The tosyl-L-phenylalinine chloromethyl ketone was not only the inhibitor of the chymotrypsin-like enzyme,but also the activator of the weak alkaline trypsin-like enzyme.Comparison between the insect protease and bovine counterpart protease revealed difference ininhibitor sensitivity.Ovomucoid trypsin inhibitor that inhibited bovine trypsinhad no effect on insect trypsin-like enzymes.Soybean trypsin inhibitor had greatinhibition on both insect and bovine proteases.
Keywords:Helicoverpa armigera  protease  activator  inhibitor  
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