02157naa a2200265 a 450000100080000000500110000800800410001910000230006024501740008326000090025752012990026665000180156565000190158365000230160265300280162565300330165370000210168670000180170770000180172570000200174370000220176370000260178570000240181177300560183519798262024-04-29       2013    bl uuuu     u00u1 u    #d1 aSILVA, M. C. M. da  aEmploying in vitro directed molecular evolution for the selection of α-amylase variant inhibitors with activity toward cotton boll weevil enzyme.h[electronic resource]  c2013  aNumerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of -amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, -amylase inhibitors (-AIs) represent a powerful tool to apply in the control of insect pests. Here, we applied DNA shuffling and phage display techniques and obtained a combinatorial library containing 108 ˛-AI variant forms. From this library, variants were selected exhibiting in vitro affinity for cotton boll weevil -amylases. Twenty-six variant sequences were cloned into plant expression vectors and expressed in Arabidopsis thaliana. Transformed plant extracts were assayed in vitro to select specific and potent -amylase inhibitors against boll weevil amylases. While the wild type inhibitors, used to create the shuffled library, did not inhibit the A. grandis -amylases, three -AI mutants, named -AIC3, -AIA11 and -AIG4 revealed high inhibitory activities against A. grandis -amylases in an in vitro assay. In summary, data reported here shown the potential biotechnology of new ˛-AI variant genes for cotton boll weevil control.  aDNA shuffling  ainsect control  aAnthonomus Grandis  aAlfa-amylase inhibitors  aIn vitro molecular evolution1 aSARTO, R. P. del1 aLUCENA, W. A.1 aRIGDEN, D. J.1 aTEIXEIRA, F. R.1 aBEZERRA, C. de A.1 aALBUQUERQUE, E. V. S.1 aGROSSI DE SA, M. F.  tJournal of Biotechnologygv. 167, p. 377-385, 2013.