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خالص‌سازی و تعیین ویژگیهای بیوشیمیایی فنلاکسیداز همولمف شبپره برگخوار توت Glyphodes pyloalis (Lep.: Pyralidae) | ||
| آفات و بیماری‌های گیاهی | ||
| مقاله 12، دوره 83، شماره 2 - شماره پیاپی 101، اسفند 1394، صفحه 209-222 اصل مقاله (489.42 K) | ||
| نوع مقاله: حشره شناسی کشاورزی | ||
| شناسه دیجیتال (DOI): 10.22092/jaep.2016.106118 | ||
| نویسندگان | ||
| محبوبه شریفی1؛ محمد قدمیاری* 2؛ رضا حسن ساجدی3 | ||
| 1دانشجوی دکتری گروه گیاه‌پزشکی دانشگاه گیلان | ||
| 2دانشیار گروه گیاه‌پزشکی دانشگاه گیلان | ||
| 3گروه بیوشیمی دانشگاه تربیت مدرس | ||
| چکیده | ||
| آنزیم فنل اکسیداز (EC 1.14.18.1)، یکی از آنزیمهای کلیدی در رشد و ایمنی حشرات میباشد و مهار این آنزیم میتواند بهعنوان روشی جدید برای کنترل آفات مورد استفاده قرار گیرد. شبپره برگخوار توت (Glyphodes pyloalis Walker)، یکی از آفات مهم درختان توت در شمال ایران است که با تغذیه از برگ توت در پرورش کرم ابریشم اختلال ایجاد میکند. در این تحقیق، آنزیم فنلاکسیداز از همولمف شبپره برگخوار توت با استفاده از رسوب با سولفات آمونیوم، کروماتوگرافی ژل و کروماتوگرافی تبادل یونی خالص شد. وزن ملکولی دو ایزوفرم آنزیم با استفاده از SDS-PAGE بهترتیب 69/66 و 70/53 کیلودالتون بهدست آمد. اثرات بازدارندهها نشان داد که روش مهارکنندگی 4-هگزیل رسورسینول از نوع رقابتی و اما برای کوئرسین هیدرات و کوجیک اسید از نوع مختلط میباشد. هم‌چنین، بررسی ویژگیهای آنزیمی نشان داد که اسیدیته مطلوب و دمای بهینه آنزیم فنلاکسیداز بهترتیب برابر با 7 و 35 درجه سلسیوس بود و آنزیم در دماهای 40، 45 و 50 درجه سلسیوس تا 30 دقیقه پایدار بود. اثر یونهای فلزی روی آنزیم خالص نشان داد که یون Zn2+ کاهشی معنی‌داری بر فعالیت آنزیم داشت. | ||
| کلیدواژه‌ها | ||
| Glyphodes pyloalis؛ فنلاکسیداز؛ خالص‌سازی؛ مکانیسم مهارکنندگی | ||
| عنوان مقاله [English] | ||
| Purification and biochemical characterization of phenoloxidase from hemolymph of Glyphodes pyloalis (Lep: Pyralidae) | ||
| نویسندگان [English] | ||
| M. SHARIFI1؛ M. GHADAMYARI2؛ R. H. SAJEDI3 | ||
| چکیده [English] | ||
| Insect phenoloxidase (PO) (EC 1.14.18.1) is the key enzyme in development and immunity of insects. Inhibition of this enzyme could be a new target for pest control. The lesser mulberry snout moth, Glyphodes pyloalis Walker is an important pest of mulberry trees in north of Iran. This pest feeds on mulberry leaves and causes serious problems for the silk industries. In this study, PO from hemolymph of G. pyloalis was purified by ammonium sulfate precipitation, ion exchange chromatography and gel filtration. The apparent molecular weights of two isoforms were determined by SDS-PAGE as 69.66 and 70.53 KDa. The inhibitory kinetics of PO showed that the mechanisms were competitive and mixed inhibition for 4-hexylresorcinol, kojic acid and quercetin, respectively. Optimal pH and maximum temperature for PO activity purified from G. pyloalis were 7 and 35 °C, respectively. The purified PO was stable at 40, 45 and 50 ºC for 30 min. The effects of different ions on enzyme activity showed that Zn2+ had a significant inhibitory effect. | ||
| کلیدواژه‌ها [English] | ||
| Glyphodes pyloalis, Phenoloxidase, PURIFICATION, Mechanism of inhibitory | ||
| مراجع | ||
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