Carbonic anhydrase from Apis mellifera: purification and inhibition by pesticides

Figures & data

Figure 1. Structures of pesticides tested for the inhibition of AmCA activity.

Figure 2. SDS-PAGE photograph. Lane 1: truncated β-galactosidase (83 kDa). Lane 2: standard proteins: truncated β-galactosidase (83 kDa), bovine albumin (66 kDa) and bovine carbonic anhydrase (29 kDa). Line 3: AmCA estimated molecular weight 37 kDa.

Table 1. Summary of purification procedure of CA from honey bee, AmCA.

Purification step	Total volume (ml)	Protein (mg/ml)	Total protein (mg)	Total activity (EU)	Spesific activity (EU/mg)	Yield (%)	Purification factor
Homogenate	5	12.1	141	705	5	100	1
Affinity Chromatography	3	0.24	0.72	126	175	17.87	35

Table 2. AmCA inhibition data with studied pesticides (IC₅₀ values), by an esterase assay with 4-nitrophenylacetate as substrate.

Inhibitor	hbCA (μM)
Tebuconazole	0.0030
Propoxur	0.0321
Carbaryl	0.0031
Carbofuran	0.0087
Simazine	0.0273
Atrazine	0.0165