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Production and Characterization of N- and C-terminally Truncated Mtx2: a Mosquitocidal Toxin from Bacillus sphaericus
Authors:Kulwadee Phannachet  Ponlawoot Raksat  Thidarat Limvuttegrijeerat  Boonhiang Promdonkoy
Affiliation:1. Department of Biotechnology, Faculty of Science and Technology, Thammasat University, 99 Pahonyothin Road, Klong 1, Klong Luang, Pathumthani, 12120, Thailand
2. National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Pahonyothin Road, Klong 1, Klong Luang, Pathumthani, 12120, Thailand
Abstract:Mosquitocidal toxin 2 (Mtx2) is a mosquito-larvicidal protein produced during vegetative stage of Bacillus sphaericus. The toxin consists of 292 amino acids and has a molecular weight of 31.8 kDa. To determine the active core region of the toxin, amino acids at N- and C-termini were sequentially removed. Deletion up to 23 amino acids from the N-terminus (Met1-Tyr23) did not significantly affect protein production and the toxin activity, whereas removal of 26 amino acids from the N-terminus (Met1-Lys26) completely abolished toxicity even though the protein production remained unchanged. Deletion of only 5 amino acids from the C-terminal end yielded the protein that could not be solubilized and rendered the toxin inactive. The results demonstrated that the C-terminal end of Mtx2 is required for proper folding and toxicity. Amino acids at the N-terminus up to Tyr23 did not play a significant role in protein production and toxicity whereas amino acids between Thr24 and Lys26 are required for full toxicity.
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