To investigate the biochemical characterization of the carboxylesterase LmCesA1 from Locusta migratoria.
We expressed recombinant LmCesA1 in Sf9 cells by using the Bac-to-bac baculovirus expression system. Enzyme kinetic assays showed that the Km values of LmCesA1 for α-naphthyl acetate (α-NA) and β-naphthyl acetate (β-NA) were 0.08 ± 0.01 mM and 0.22 ± 0.03 mM, respectively, suggesting that LmCesA1 has a higher affinity for α-NA. LmCesA1 retained its enzymatic activity during incubations at pH 7–10 and at 10–30 °C. In an inhibition experiment, two organophosphate pesticides (malaoxon and malathion) and one pyrethroid pesticide (deltamethrin) showed different inhibition profiles against purified LmCesA1. Recombinant LmCesA1 activity was significantly inhibited by malaoxon in vitro. UPLC analysis showed that no metabolites were detected.
These results suggest that overexpression of LmCesA1 enhances malathion sequestration to confer malathion tolerance in L. migratoria.
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This research was supported by the National Natural Science Foundation of China (International Cooperation and Exchange Programme, No. 30810103907), Programs of Applied Basic Research of Shanxi Province (No. 201601D202058) and 2016 Provincial Support National Research Foundation of Shanxi Province (No. 226546001). The authors are grateful to Prof. Yoonseong Park and Rupinder Singh at the Department of Entomology, Kansas State University, Manhattan, Kansas, USA, for critical reading of manuscript.
Supplementary Table 1—Standard curve of CarE activity.
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Yin, F., Ma, W., Li, D. et al. Expression and kinetic analysis of carboxylesterase LmCesA1 from Locusta migratoria. Biotechnol Lett (2021). https://doi.org/10.1007/s10529-021-03086-1
- Bac-to-bac baculovirus expression system
- Locusta migratoria