TAILIEUCHUNG - Báo cáo khoa học: Digestive a-amylases of the flour moth Ephestia kuehniella – adaptation to alkaline environment and plant inhibitors

The digestive tract of lepidopteran insects is extremely alkaline. In the pres-ent work, molecular adaptation of amylolytic enzymes to this environment was investigated in the flour moth Ephestia kuehniella, an important stored-product pest. Three digestivea-amylases [Ephestia kuehniellaa-amy-lase isoenzymes 1–3 (EkAmy1–3)] with an alkaline pH optimum were puri-fied from larvae and biochemically characterized. | ỊFEBS Journal Digestive a-amylases of the flour moth Ephestia kuehniella - adaptation to alkaline environment and plant inhibitors 12 1 1 3 2 Jana Pytelkova Jan Hubert Martin Lepsik Jan Sobotnik Radek Sindelka Iva KriZkova Martin Horn1 and Michael Mares1 1 Institute of Organic Chemistry and Biochemistry AS CR . Praha Czech Republic 2 Research Institute of Crop Production . Praha Czech Republic 3 Institute of Biotechnology AS CR . Praha Czech Republic Keywords alkaline adaptation a-amylase a-amylase inhibitor Ephestia kuehniella plant-insect interaction Correspondence M. Mares Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic Flemingovo nam. 2 166 10 Praha 6 Czech Republic Fax 420 220183578 Tel 420 220183358 E-mail mares@ Received 12 January 2009 revised 18 March 2009 accepted 24 April2009 doi The digestive tract of lepidopteran insects is extremely alkaline. In the present work molecular adaptation of amylolytic enzymes to this environment was investigated in the flour moth Ephestia kuehniella an important stored-product pest. Three digestive a-amylases Ephestia kuehniella a-amy-lase isoenzymes 1-3 EkAmy1-3 with an alkaline pH optimum were purified from larvae and biochemically characterized. These isoenzymes differ significantly in their sensitivity to a-amylase inhibitors of plant origin that are directed against herbivores as antifeedants. Such functional variability renders the amylolytic system less vulnerable to suppression by plant defensive molecules. Moreover we found that expression of a-amylases is upreg-ulated in larvae feeding on a diet enriched with an a-amylase inhibitor. The a-amylases are secreted into the larval midgut by an exocytotic mechanism as revealed by immunogold microscopy. The cDNA sequence of EkAmy3 was determined and a homology model of EkAmy3 was built in order to analyze the structural features responsible for adaptation to alkaline .

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