Date of this Version
Published in Comparative Biochemistry and Physiology, Part B 139 (2004), pp. 27–33; doi: 10.1016/j.cbpc.2004.05.010.
We describe a phospholipase A2 (PLA2) associated with the salivary glands of tobacco hornworms, Manduca sexta. This enzyme is able to hydrolyze arachidonic acid from the sn-2 position of PLs. Addition of the calcium chelator, EGTA, or calcium, to the Tris reaction buffer impaired the PLA2 activity, from which we infer the enzyme requires very low concentrations of calcium or perhaps other ions for optimal activity. PLA2 activity was sensitive to protein concentration (highest activity at 25 μg protein per μl), reaction time (optimal at 30 min), buffer pH (optimal at pH 8–10), and reaction temperature (optimal range 18–38°C). The salivary gland PLA2 was sensitive to the site-specific inhibitor, oleyloxyethylphosphorylcholine and stable to freezing at –80°C, but not –20°C. The biological significance of this enzyme may relate to hydrolysis of fatty acid moieties from dietary PLs for absorption by midgut epithelia. This salivary gland enzyme may also be responsible for killing food-borne bacteria.