ORIGINAL ARTICLE
Isolation and purification of α-amylase inhibitors and their in vitro and in vivo effects on Tribolium castaneum (Herbst) and Callosobruchus maculatus (F.)
 
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1
Department of Plant Protection, Faculty of Agriculture, Sohag University, Sohag, Egypt
2
Physico-Chemical Biology, A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article
CORRESPONDING AUTHOR
Ashraf Oukasha Abd El-latif   

Department of Plant Protection, Faculty of Agriculture, Sohag University, Sohag, Egypt
Submission date: 2020-07-29
Acceptance date: 2020-08-19
Online publication date: 2020-12-02
 
Journal of Plant Protection Research 2020;60(4):377–387
 
KEYWORDS
TOPICS
ABSTRACT
Plant derived α-amylase inhibitors are proteinaceous molecules that regulate the enzyme activity in plants and also protect plants from insect attack. In the current study, 28 accessions of 19 plant species were screened for their α-amylase inhibitory activity. The durum wheat varieties, Beni Suef-1 and Beni Suef-5, showed strong α-amylase inhibitory activity and were subjected to further purification studies using ammonium sulfate fractionation and DEAE-Sephadex G-25 column. The isolated inhibitors were found to be stable at temperatures below 80°C with maximum activity obtained at 40−50°C. Also, they were stable in a wide pH range (2−12). The ion exchange products of purified α-amylase inhibitors from Beni Suef-1 and Beni Suef-5 varieties showed a molecular weight of 16 and 24 kDa, respectively. The purified α-amylase inhibitors were tested against Tribolium castaneum and Callosobruchus maculatus both in vitro and in vivo. There was linear inhibition of α-amylase activity with increasing inhibitor concentration until saturation was reached. Beni Suef-5 α-amylase inhibitor was more potent against α-amylase with lower IC50 values than Beni Suef-1 α-amylase inhibitor except in the case of T. castaneum larva. Kinetics analysis revealed that Beni Suef-1 and Beni Suef-5 α-amylase inhibitors are non- -competitive types of inhibitors with high affinity toward α-amylase of T. castaneum and C. maculatus. Results of the in vivo studies demonstrated that α-amylase inhibitors isolated from durum wheat, Beni Suef-1 and Beni Suef-5 varieties, were very effective in inhibiting the development of T. castaneum and C. maculatus and could be used for future studies in developing insect resistant transgenic plants approaching α-amylase inhibitor genes.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
FUNDING
The financial support (project 26601) provided by the Science and Technology Development Fund (STDF), Egypt is fully acknowledged.
 
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