Characterization and inhibitor studies of chitinases from a parasitic blowfly (Lucilia cuprina), a tick (Boophilus microplus), an intestinal nematode (Haemonchus contortus) and a bean (Phaseolus vulgaris)

Article abstract of DOI:10.1002/(SICI)1096-9063(199612)48:4<305::AID-PS480>3.0.CO;2-I

The molecular weight pattern and the stage-specific activities of chitinases from the blowfly Lucilia cuprina, the tick Boophilus microplus and the intestinal nematode Haemonchus contortus were examined. Chitinolytic enzymes could be detected in all parasite species tested, but the activity was different between the stages. Highest chitinolytic titers were found in blowfly pupae (83 kDa, 118 kDa), hatching larvae of ticks (58 kDa, 94 kDa) and nematode eggs (43 kDa). Leaves from ethylene-treated bean plants Phaseolus vulgaris expressed two basic Class I chitinases (Ia, Ib) of 34 kDa, differing in their amino acid sequences at residue 33 and 34 (Ia: glycine, proline; Ib: lysine, aspartic acid). Inhibitor studies with blowfly pupae revealed that allosamidin (IC₅₀ = 0.32 (±0.02) μM) was by far the best inhibitor when compared with various amino sugar derivatives. This compound also inhibited chitinases from tick larvae (IC₅₀ = 0.69(±0.10) μM) and nematode eggs (IC₅₀ = 0.048(±0.0045) μM) specifically. Whereas Class Ia chitinase from bean leaves was inhibited only up to 18% by 10 μM allosamidin, it had an IC₅₀ of 1(±0.14) μM for the Ib type, which is the first plant chitinase described to be highly sensitive to allosamidin.