Premium
Effect of adding amino acids residues in N‐ and C‐terminus of Vip3Aa16 (L121I) toxin
Author(s) -
Sellami Sameh,
Cherif Marwa,
Jamoussi Kaïs
Publication year - 2016
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201500712
Subject(s) - escherichia coli , amino acid , proteolysis , n terminus , protease , biology , biochemistry , bacillus thuringiensis , c terminus , peptide , toxin , fusion protein , peptide sequence , mutant , bacteria , enzyme , recombinant dna , gene , genetics
To study the importance of N‐ and C‐terminus of Bacillus thuringiensis Vip3Aa16 (L121I) toxin (88 kDa), a number of mutants were generated. The addition of two (2R: RS) or eleven (11R: RSRPG) amino acid residues at the Vip3Aa16 (L121I) C‐terminus allowed to an unappropriated folding illustrated by the abundant presence of the 62 kDa proteolytic form. The produced Vip3Aa16 (L121I) full length form was less detected when increasing the number of amino acids residues in the C‐terminus. Bioassays demonstrated that the growth of the lepidopteran Ephestia kuehniella was slightly affected by Vip3Aa16 (L121I)‐2R and not affected by Vip3Aa16 (L121I)‐11R. Additionally, the fusion at the Vip3Aa16 (L121I) N‐terminus of 39 amino acids harboring the E. coli OmpA leader peptide and the His‐tag sequence allowed to the increase of protease sensitivity of Vip3Aa16 (L121I) full length form, as only the 62 kDa proteolysis form was detected. Remarkably, this fused protein produced in Escherichia coli ( E. coli ) was biologically inactive toward Ephestia kuehniella larvae. Thus, the N‐terminus of the protein is required to the accomplishment of the insecticidal activity of Vip3 proteins. This report serves as guideline for the study of Vip3Aa16 (L121I) protein stability and activity.