Purification and Biochemical Properties of Extracellular Phospholipase $A_1$ from $Serratia$ sp. MK1

Abstract

A novel type of extracellular phospholipase A(1) was isolated from Serratia sp. MK1 and purified to homogeneity by ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The purified enzyme was a monomer with a molecular mass of about 43,000 Da. This enzyme showed the highest lipolytic activity toward phosphatidylserine among the phosphoglycerides tested, and preferentially catalyzed the hydrolysis of the ester bond in phosphatidic acid to lyso-phosphatidic acid. Enzyme activity was completely inhibited by the addition of a chelating agent such as EDTA, and inhibited enzyme activity was fully recovered by the presence of Ca2+. This implies that the enzyme requires Ca2+ for activity. The enzyme was stable up to 70 degrees C when incubated for 1 h at pH 8.5, and the optimal pH and temperature were 8.5 and 50 degrees C, respectively.