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棉铃虫酯酶两个位点突变对三种杀虫剂代谢活性的影响

, PP. 271-276

Keywords: 酯酶,氨基酸突变,有机磷,顺式氰戊菊酯,代谢活性

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Abstract:

为研究棉铃虫酯酶对有机磷和拟除虫菊酯类杀虫剂的代谢活性,在前期对酯酶001F和001G酶活性研究的基础上,进一步采用荧光法和高效液相色谱法测定了酯酶及其突变体(A127位和F238位突变)对二甲基-4甲基伞形酮磷酸酯(dMUP)、二乙基-7羟基香豆素磷酸酯(dECP)及顺式氰戊菊酯(2S,αS)的酶促代谢活性。酯酶001F和001G对有机磷的酶促反应常数(kcat值)在2.0×10-3/min~5.2×10-3/min之间,后者对顺式氰戊菊酯的代谢活性为10.19μmol?min-1?μmol-1;001F的突变提高了对有机磷的代谢活性,其中001FA127D和001FF238L对dMUP的kcat值至少是突变前的2倍;001G的突变却降低了对dECP的代谢活性,但001GF238L对dMUP的活性有所提高。此外,突变体中001FA127D对顺式氰戊菊酯的活性与突变前接近,而001GF238L则降低到突变前的1/2,其它突变体未检测到活性。结果表明,酯酶001F和001G对杀虫剂有代谢活性,A127位和F238位的突变对酶活性有显著影响。

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