Purification and properties of two phospholipases D from Streptomyces sp.
Two enzymes with phospholipase D activity were purified from Streptomyces strains (PMF and PM43) by column chromatography on Fractogel TSK CM-650(S), Sephadex G-100 and Fractogel EMD DEAE-650(M). The purified preparations were found to be homogeneous by SDS-PAGE, capillary electrophoresis and analytical gel filtration. The molecular masses, assessed by MALDI-MS spectrometry, were 53.864 kDa for PMF and 54.147 kDa for PM43. The isoelectric point was 9.1 for both enzymes. The enzymes were most active at around 60 degrees C and stable between pH 4 and 9 and below 50 degrees C. The pH optima were between 4 and 6 for PMF and between 6 and 7 for PM43. Both phospholipases displayed high transphosphatidylation activity but PMF was more selective than PM43.
Carrea
G
G
CNR, Istituto di Chimica degli Ormoni, Milano, Italy.
D'Arrigo
P
P
Piergianni
V
V
Roncaglio
S
S
Secundo
F
F
Servi
S
S
D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D010739:Phospholipase D / Q000737:chemistry / Q000302:isolation & purification*; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D013696:Temperature
D007527:Isoenzymes; D010739:Phospholipase D