Phosphatidylinositol-specific phospholipase C from Bacillus cereus: improved purification, amino acid composition, and amino-terminal sequence.
Phosphatidylinositol-specific phospholipase C was purified in a 27% yield from the culture medium of Bacillus cereus by a combination of ammonium sulfate precipitation and ion-exchange and hydrophobic interaction chromatography. The purified enzyme was free of other phospholipase C-type activities and exhibited a high specific activity of approximately 1,300 units/mg. Amino acid composition analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a molecular weight of about 35 kDa. The sequence of the first 29 N-terminal amino acids was also determined.
Volwerk
J J
JJ
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Wetherwax
P B
PB
Evans
L M
LM
Kuppe
A
A
Griffith
O H
OH
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D001409:Bacillus cereus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism*; D010738:Type C Phospholipases / Q000302:isolation & purification*
D000596:Amino Acids; D010716:Phosphatidylinositols; D010738:Type C Phospholipases