02323nas a2200265   4500008004100000022001400041245013000055210006900185260000800254300001200262490000700274520144300281653001201724653001301736653002401749653001801773653001501791653001201806653001701818100002201835700002601857700002201883700002001905856013201925       2023                        eng d  a1572-388700aDomain shuffling and site-saturation mutagenesis for the enhanced inhibitory potential of amaranthaceae α-amylase inhibitors0 aDomain shuffling and sitesaturation mutagenesis for the enhanced  cOCT  a519-5320 v423 a<p>
	Amaranthaceae a-amylase inhibitors (AAIs) are knottin-type proteins with selective inhibitory potential against coleopteran a-amylases. Their small size and remarkable stability make them exciting molecules for protein engineering to achieve superior selectivity and efficacy. In this report, we have designed a set of AAI pro- and mature peptides chimeras. Based on in silico analysis, stable AAI chimeras having a stronger affinity with target amylases were selected for characterization. In vitro studies validated that chimera of the propeptide from Chenopodium quinoa a-AI and mature peptide from Beta vulgaris a-AI possess 3, 7.6, and 4.26 fold higher inhibition potential than parental counterparts. Importantly, recombinant AAI chimera retained specificity towards target coleopteran a-amylases. In addition, to improve the inhibitory potential of AAI, we performed in silico site-saturation mutagenesis. Computational analysis followed by experimental data showed that substituting Asparagine at the 6th position with Methionine had a remarkable increase in the specific inhibition potential of Amaranthus hypochondriacus a-AI. These results provide structural-functional insights into the vitality of AAI propeptide and a potential hotspot for mutagenesis to enhance the AAI activity. Our investigation will be a toolkit for AAI's optimization and functional differentiation for future biotechnological applications.</p>
10a& alpha10a-Amylase10a-Amylase inhibitors10aAmaranthaceae10aColeoptera10aPeptide10aPest control1 aRane, Ashwini, S.1 aNair, Vineetkumar, S.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.  uhttps://library.ncl.res.in/content/domain-shuffling-and-site-saturation-mutagenesis-enhanced-inhibitory-potential-amaranthaceae