Facile One-Pot Immobilization of a Novel Thermostable Carboxylesterase from Geobacillus uzenensis for Continuous Pesticide Degradation in a Packed-Bed Column Reactor

The novel carboxylesterase gene (est741) was cloned from Geobacillus uzenensis. The optimal pH and temperature of Est741 were 8.0 and 50 °C. Through site-directed mutation, the optimum temperature of the mutant M160K(EstM160K) was increased from 50 to 60 °C, and showed enhanced T1/2 of 2.5 h at 70 °C in comparison to the wild type (1.3 h). EstM160K was successfully expressed Pichia pastoris and EstM160K fermentation broth was directly immobilized on epoxy-functionalized supports via a one-pot strategy to obtain the immobilized enzyme lx-EstM160K. Additionally, lx-EstM160K showed enhanced T1/2 of 36.8 h at 70 °C in comparison to free enzyme. lx-EstM160K could degrade various pyrethroid pesticides. After 40 min reaction with 50 U of the lx-EstM160K, the malathion removal was 95.8% with a malathion concentration of 20 mg/L. When 2.5 g lx-EstM160K was added to the 10 mL column reactor with the concentration of bifenthrin was 500 mg/L and the transfer rate of the pump was 0.7 mL/min, the degradation rate of lx-EstM160K to bifenthrin was 90.4%. lx-EstM160K exhibited high operational stability and maintained 72% initial activity after ten batches of continuous reaction for bifenthrin pesticide biodegradation.

Edaphic Entomofauna Variation Depending on Glyphosate Application in Roundup Ready Soybean Crops

ABSTRACT: Currently, biomonitoring is a methodology used to verify the environmental impact of new technologies in the agricultural environment, highlighting edaphic entomofauna that is traditionally used as a bioindicator in this sort of research. Therefore, the objective of this study was to investigate the edaphic entomofauna variation depending on glyphosate application in Roundup Ready soybeans. The experiment was carried out in Coimbra, MG during the 2007/2008 and 2008/2009 cropping seasons. The experimental design was a randomized block design with five replications. The treatments were: non-transgenic soybean with mechanical weeding of weeds; transgenic soybean with mechanical weeding of weeds; transgenic soybean with one glyphosate application and transgenic soybean with three glyphosate applications. The populations of the edaphic entomofauna were sampled during two crops. The insertion of the glyphosate tolerance gene did not affect the richness and the abundance of arthropods in the soil. The arthropod richness was reduced in treatments where glyphosate was applied one and three times. The glyphosate application in transgenic soybean reduced the density of the predatory mite Galumnidae (Acari); predator ants Neivamyrmex sp. (Hymenoptera: Formicidae) and Solenopsis sp. (Hymenoptera: Formicidae); and springtails Entomobryidae (Collembola), Hypogastrura sp. (Collembola: Hypogastruridae) and Onychiuridae (Collembola). Therefore, it is essential to follow the use recommendations of the herbicide glyphosate and adopt good agricultural practices that promote pesticide biodegradation, thereby contributing to the reduction of the toxicological potential of glyphosate on the edaphic entomofauna.