| Title : Bacterial Expression and Kinetic Analysis of Carboxylesterase 001D from Helicoverpa armigera - Li_2016_Int.J.Mol.Sci_17_ |
| Author(s) : Li Y , Liu J , Lu M , Ma Z , Cai C , Wang Y , Zhang X |
| Ref : Int J Mol Sci , 17 : , 2016 |
| Abstract : Carboxylesterasesare an important class of detoxification enzymes involved in insecticide resistance in insects. A subgroup of Helicoverpa armigera esterases, known as Clade 001, was implicated in organophosphate and pyrethroid insecticide resistance due to their overabundance in resistant strains. In this work, a novel carboxylesterasegene 001D of H. armigera from China was cloned, which has an open reading frame of 1665 nucleotides encoding 554 amino acid residues. We used a series of fusion proteins to successfully express carboxylesterase 001D in Escherichia coli. Three different fusion proteins were generated and tested. The enzyme kinetic assay towards 1-naphthyl acetate showed all three purified fusion proteins are active with a Kcat between 0.35 and 2.29 s(-1), and a Km between 7.61 and 19.72 muM. The HPLC assay showed all three purified fusion proteins had low but measurable hydrolase activity towards beta-cypermethrin and fenvalerate insecticides (specific activities ranging from 0.13 to 0.67 muM.min(-1).(muM(-1).protein)). The enzyme was stable up to 40 degrees C and at pH 6.0-11.0. The results imply that carboxylesterase 001D is involved in detoxification, and this moderate insecticide hydrolysis may suggest that overexpression of the gene to enhance insecticide sequestration is necessary to allow carboxylesterases to confer resistance to these insecticides in H. armigera. |
| ESTHER : Li_2016_Int.J.Mol.Sci_17_ |
| PubMedSearch : Li_2016_Int.J.Mol.Sci_17_ |
| PubMedID: 27049381 |
| Gene_locus related to this paper: helam-d5kxa9 |
| Gene_locus related to this paper: helam-d5kxa9 |
Li Y, Liu J, Lu M, Ma Z, Cai C, Wang Y, Zhang X (2016)
Bacterial Expression and Kinetic Analysis of Carboxylesterase 001D from Helicoverpa armigera
Int J Mol Sci
17 :
Li Y, Liu J, Lu M, Ma Z, Cai C, Wang Y, Zhang X (2016)
Int J Mol Sci
17 :