Influence of Coprecipitation and Interactions with Anionic Surfactants on the Activity of Plant Phospholipase D

E. V. Shumilina, N. Yu. Zonova, O. G. Mukhaneva, and Yu. A. Shchipunov^*

Institute of Chemistry, Far East Branch of the Russian Academy of Sciences, Vladivostok, 690022 Russia; fax: (4232) 311-889; E-mail: chemi@online.ru

^* To whom correspondence should be addressed.

Received October 29, 1998; Revision received March 23, 1999

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The effects of two surfactants (sodium dodecyl sulfate and sodium dodecyl dioxyethylene sulfate) of similar structure but differing water solubility (of their calcium salts) on the enzymatic activity of cabbage phospholipase D have been studied. The solubility difference is insignificant because the two surfactants activate phospholipase D similarly. To elucidate the mechanism of their influence on the enzyme, the phase behavior in the reaction media and the interactions of the surfactants with the enzyme were investigated by potentiometry and by light scattering and UV spectroscopy. Calcium dodecyl dioxyethylene sulfate (which is more soluble in water than calcium dodecyl sulfate) precipitates in the presence of phosphatidylcholine, the substrate of the enzymatic reaction. In the reaction media phospholipase D was involved into a precipitate consisting of calcium salts of the surfactants and phosphatidylcholine that might be interpreted as its immobilization. In addition, the surfactants were adsorbed on the enzyme, unfolding the globular enzyme molecule due to electrostatic repulsion between adsorbed surfactant anions. The observed increase in the functional activity of phospholipase D is accounted for by transfer to an optimal tertiary structure for the enzyme molecule in the course of consecutive conformational transitions induced by the surfactants.

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KEY WORDS: phospholipase D, enzymatic activity, surface-active substances, coprecipitation, interactions, tertiary structure

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