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PEDS Advance Access originally published online on November 29, 2007
Protein Engineering Design and Selection 2007 20(12):599-606; doi:10.1093/protein/gzm058
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© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Prediction-based protein engineering of domain I of Cry2A entomocidal toxin of Bacillus thuringiensis for the enhancement of toxicity against lepidopteran insects

Chandi C. Mandal1,3, Srimonta Gayen1, Asitava Basu1, Kalyan S. Ghosh2, Swagata Dasgupta2, Mrinal K. Maiti1 and Soumitra K. Sen1,4

1IIT – BREF Biotek, Indian Institute of Technology, Kharagpur-721 302, India 2Department of Chemistry, Indian Institute of Technology, Kharagpur-721 302, India

4 To whom correspondence should be addressed. E-mail: sk_sen55{at}yahoo.co.in

Issues relating to sustenance of the usefulness of genetically modified first generation Bt crop plants in the farmer’s field are of great concern for crop scientists. Additional biotechnological strategies need to be in place to safeguard the possibility for yield loss of Bt crop by other lepidopteran insects that are insensitive to the Cry1A toxin, and also against the possibility for emergence of resistant insects. In this respect, Cry2A toxin has figured as a prospective candidate to be the second toxin to offer the required protection along with Cry1A. In the present study, the entomocidal potency of Cry2A toxin was enhanced through knowledge-based protein engineering of the toxin molecule. Deletion of 42 amino acid residues from the N-terminal end of the peptide followed by the replacement of Lys residues by nonpolar amino acids in the putative transmembrane region including the introduction of Pro resulted in a 4.1–6.6-fold increase in the toxicity of the peptide against three major lepidopteran insect pests of crop plants.

Keywords: Bacillus thuringiensis {delta}-endotoxins/enhancement of insecticidal property/insect resistance management/protein engineering of Cry2A/transmembrane domain

Received April 2, 2007; revised August 28, 2007; accepted September 27, 2007.

3 Present address: Department of Pathology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78 229, USA


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