Pesticide application inhibit the microbial carbonic anhydrase-mediated carbon sequestration in a soil microcosm

Heterotrophic system for carbon sequestration is gaining importance in the recent decades. Carbonic anhydrase (CA) is a major enzyme involved in carbon sequestration and biomineralization process. In this paper, we evaluate the effect of pesticide on CA activity using inhibitory assay. 2,4-D, being one of the most extensively used pesticide, being deleterious to soil health, its usage should be minimized to regain the soil health. Maximum inhibitory constant (K_i) was observed for 5% 2,4-D (49.53 mM) followed by 5% glyphosate (43.92 mM). The maximum Km increase with increase in pesticide concentration by 3.05-fold was in case of glyphosate which was higher than that of 2,4-D (2.08-fold) and dichlorvos (2.38-fold). Moreover, we evaluated the carbon sequestration using CA enzyme in the soil microcosm. In the present study, we identified the negative impact of 2,4-D on carbonic anhydrase produced by Bacillus halodurans PO15. The inhibition was a mixed type and had significantly lowered the carbon reduction to about 2.38 ± 0.17% in a soil microcosm study. Based on the molecular docking, the inhibition was contributed due to weak H-bonding interaction with amino acid residues (Gly65, Gly95, Val147, Ser150 and Gly65, Ser146, and Ser150).