title: Purification and properties of two phospholipases D from Streptomyces sp.
abstract: 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.
authors.affiliation: CNR, Istituto di Chimica degli Ormoni, Milano, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Carrea
authors.forename: P
authors.initials: P
authors.lastname: D'Arrigo
authors.forename: V
authors.initials: V
authors.lastname: Piergianni
authors.forename: S
authors.initials: S
authors.lastname: Roncaglio
authors.forename: F
authors.initials: F
authors.lastname: Secundo
authors.forename: S
authors.initials: S
authors.lastname: Servi
mesh_terms: 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
chemical_list: D007527:Isoenzymes; D010739:Phospholipase D