A. Abousalham et al., IMPROVED PURIFICATION AND BIOCHEMICAL-CHARACTERIZATION OF PHOSPHOLIPASE-D FROM CABBAGE, Biochimica et biophysica acta, 1158(1), 1993, pp. 1-7
Phospholipase D (phosphatidylcholine phosphatidohydrolase, EC 3.1.4.4)
was purified from cabbage leaves. The two step purification procedure
involved hydrophobic chromatography on Octyl-Sepharose followed by a
Mono-Q/FPLC-column with a total yield of 23% and a purification factor
of 1000. A zymographic assay was used to detection of PL D activities
at various stages of purification under non denaturing PAGE. The mole
cular mass was determined to be 90 kDa using the SDS/PAGE method, and
90 200 Da as calculated from the amino acid analysis. The isoelectric
point of the enzyme is acidic (pI = 4.7). The amino-acid composition a
nd 29 residues of the NH2-terminal amino-acid sequence were determined
.