Purification and characterization of an esterase isozyme from insecticide resistant and susceptible strains of German cockroach, Blattella germanica (L.).

The most active forms of esterases (E5, E6 and E7) from the German cockroach, Blattella germanica (L.) were purified from resistant and susceptible strains. About 45-155 fold purification with a 11-16% of total esterase recovery was achieved after different column chromatography and preparative gel electrophoresis. Elution profiles of resistant and susceptible strains were similar, but esterase E6 activity was higher in the resistant strains. Kinetic analyses indicate no differences in Km values between the resistant and susceptible strains. However Vmax was significantly higher in resistant strains. Inhibition of esterase activity by paraoxon, chlorpyrifos and propoxur did not suggest any structural differences in esterase E6 between strains. From these results we suggest that insecticide resistance in German cockroach is due to the increased production of E6 esterase. The role of E6 may be sequestration of toxic molecules rather than hydrolysis.

Subcellular distribution and characterization of esterase isozymes from insecticide-resistant and -susceptible strains of German cockroach (Dictyoptera: Blattellidae).

Esterases from insecticide-resistant and -susceptible strains of German cockroach, Blattella germanica (L.), were isolated and analyzed for differences in activity. The isozymes E5, E6, E7, and E8 of resistant strains indicated about two- to fivefold increases in activity compared with those of the susceptible strain. These isozymes constituted approximately 60% of the total activity in the resistant strains, whereas they contributed only about 40% activity in the susceptible strain. Subcellular distribution studies showed that most esterase activity is present in the 100,000 x g fraction of the homogenate (cytosolic). Only a small portion of activity was membrane bound. Molecular weights of the esterase isozymes calculated based on their retardation coefficients ranged between 48 and 81 kd.

Activity and electrophoretic characterization of esterases in insecticide-resistant and susceptible strains of German cockroach (Dictyoptera: Blattellidae).

Strains of German cockroach, Blattella germanica (L.) were tested for resistance to three insecticides using the time-mortality response technique in comparison with known susceptible strains. Dursban-R and Macy strains indicated high levels of resistance to chlorpyrifos and moderate resistance to propoxur and cypermethrin. The hydrolytic activity of esterase enzymes was determined with a model substrate, p-nitrophenyl acetate. The specific activity of the enzymes increased with a corresponding increase in substrate concentration. Maximum activity was observed at 80 microM. The resistant strains had significantly higher activity than the susceptible strains. The enzyme activity also increased with an increase in pH. Highest esterase activity was observed at pH 8.0 for all the strains. The calculated Michaelis Menten constant (Km) values for the resistant strains were nearly two-fold higher than for the susceptible strains. Nondenaturing gel electrophoresis revealed differences in isozyme composition. In total, 10 esterase bands were detected, and these bands were characterized using different inhibitors and substrates.