A Novel Ca2+-Dependent Phospholipase D from Streptomyces tendae, Possessing Only Hydrolytic Activity

다국어 초록 (Multilingual Abstract)

An extracellular phospholipase D (PLDSt) was purified from Streptomyces tendae by two successive chromatographic steps on Sepharose CL-6B and DEAE-Sepharose CL-6B. Molecular weight of the PLDSt was estimated to be approximately 43 kDa by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Maximal activity was at pH 8 and 60oC, and the enzyme
was stable at or below 60oC and between pH 8 and 10, when assayed after 1.5 and 24 h, respectively. The enzyme activity had an absolute requirement of Ca2+, and the maximum activity was at 2 mM CaCl2. The Km and Vmax values for phosphatidyl choline were 0.95 mM and 810 μmol min-1 mg-1, respectively. More importantly, PLDSt could not catalyze transphosphatidylation of glycerol, L-serine, myo-inositol and ethanolamine, which have been extensively
used to evaluate the activity. The result strongly suggests that PLDSt does not have the transphosphatidylation activity, thereby making it the first Streptomyces PLD possessing only hydrolytic activity. PLDSt may therefore be a novel type of PLD enzyme.

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