PURIFICATION AND N-TERMINAL SEQUENCE-ANALYSIS OF STREPTOMYCES-CHROMOFUSCUS PHOSPHOLIPASE-D

Partially purified commercial phospholipase D (PLD) was fractionated b y dye-ligand affinity chromatography and nondenaturing polyacrylamide electrophoresis (PAGE). Active material migrated as three bands on SDS -PAGE. The two higher-abundance species were shown to have identical N -terminal sequences, while the third band was present in much smaller amounts and had a distinct sequence. Cloning Streptomyces chromofuscus PLD will allow the construction of stable transfectants of mast cell lines permitting regulated expression of PLD.