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研究生:江史雯
研究生(外文):Shih-Wen Chiang
論文名稱:黑尾葉蟬羧酸酯�A的純化與特性
論文名稱(外文):Purification and characterization of carboxylesterases of a rice green leafhopper Nephotettix cincticeps
指導教授:孫志寧孫志寧引用關係
指導教授(外文):Chih-Ning Sun
學位類別:碩士
校院名稱:國立中興大學
系所名稱:昆蟲學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:1995
畢業學年度:83
語文別:中文
論文頁數:57
中文關鍵詞:黑尾葉蟬羧酸酯純化殺蟲劑
外文關鍵詞:green leafhoppercarboxylesterase isozymespurificationsecticides
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本工作利用膠體過濾以及焦集層析法,黑尾葉蟬純化出活性較高的四個
carboxylesterase isozymes, E1, E2, E3 和 E4,pI值依序為 5.1,
5.0, 4.9 和 48.5,其中 E1 和 E3 分子量大小分別為 83.7 和
58.6kDa, E2 和 E4 則包含了83.7, 58.6和42kDa 3 個蛋白質。利用
Sephadex G-75 和 Sephacryl S-2000 膠體過濾層析, 仍未能進一步將
E2 和E4中的蛋白質分離。這些isozymes對一些標準受質具有相當高的活
性表現,其中水解a-NB 的能力比 a-NA, b-NA 和 p-NPA 強, 且以E3 表現
的活性最強;而對malathion, trans-permethrin,cis-permethrin和
cypermethrin 等殺蟲劑的性較標準受質低很多,約只有水解標準受質
的1/1000000。所有isozymes 代謝除蟲菊的速率皆較 malathion 快, 且
水解 cis-permethrin 的速率較 trans-permethrin高出 1.7 至 5.0
倍; 在這些 isozymes 中, 以 E3代謝除蟲菊的能力最差, 以 E4 最高,
而且E4可以與褐飛蝨esterase抗血清有免疫交互作用。田間品系黑尾葉蟬
羧酸酯 對標準受質的活性,比感性品系高出25-37倍,但這二個品系對上述
四種殺蟲劑的代謝能力則沒有明顥差別。利用[1,3-3H]diisopropyl
fluorophosphate 標定 carboxylesterase 的結果,顥示田間品系黑尾葉
蟬分子量為 58.6kDa 的羧酸酯的量比感性品系至少高出8倍。

A combination of ammonium sulfate fractionation, gel
filtrationnd chromatofocusing was used to isolate and purify
carboxylesterases(EC3.1.1.1)of a rice green leafhopper,
Nephotettix cincticeps Uhler (Homoptera: Cicadellidae). Four
fractions, E1, E2, E3 and E4, were obtained with pIs ranging
from 5.1 to 4.85. While E1 and E3 with molecular weight of 83.7
and 58.6kDa, respectively, appeared electrophoretically
homogeneous on SDS/PAGE gel, three proteins of molecular weight
of 83.7, 58.6 and 42kDa were observed in both E2 and E4.
Further attempts using Sephadex G-75 and Sephacryl S-200
chromatography failed to resolve them. All four fractions or
isozymes exhibited significant activity toward a number of
model substrates, and a-naphthyl butyrate was hydrolyzed to
somewhat greater extent than a-naphthyl acetate, b-naphthyl
acetate and p-nitrophenyl acetate; and E3 was the most active.
Their activity toward malathion, trans-permethrin, cis-
permthrin and cypermethrin was considerably lower than their
activity toward the model substrates. Pyrethroids were
hydrolyzed faster than malathion by all carboxylesterase
isozymes, and cis-permethrin was more preferred than trans-
permethrin. E3 was in general less effective in hydrolyzing
pyrethroids than the other isozymes. E4 showed considerably
higher activity toward both permethrin and cypermethrin; and it
was the only isozyme cross-reacted with the antiserum agains E1
of the rice brown planthopper, Nilaparvata lugens. The latter
was the most active carboxylesterase isozyme showing
considerable permethrin-hydrolyzing activity in this
planthopper.

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