A novel extracellular phospholipase C purified from a marine bacterium, Pseudoalteromonas sp. J937.
Marine bacterial isolates were screened for phospholipase C (PLC) activity on PCY agar plates containing phosphatidylcholine (PC) as substrate. The strain that showed the highest activity on a PCY screening agar plate and a thin-layer chromatography was identified as a strain of Pseudoalteromonas and subsequently designated Pseudoalteromonas sp. J937. The extracellular PLC of the strain J937 was purified to a specific activity of 33 U mg(-1) protein by serial ion exchange and gel filtration column chromatography. It had a molecular mass of 32 kDa estimated by SDS-PAGE. The optimal pH and temperature of the enzyme were about pH 8 and 45 degrees C, respectively. The PLC hydrolyzed phosphatidylethanolamine as well as PC but not other glycerophospholipids. Its activity was enhanced by 150% with Ca2+ (200 mM) and by 180% with Na+ (500 mM), suggesting that the purified PLC is a marine-type enzyme.
Mo
Sangjoon
S
Division of Nano Science, Ewha Woman's University, Seoul 120-750, South Korea.
Kim
Jae-Heon
JH
Cho
Ki Woong
KW
D004795:Enzyme Stability / Q000187:drug effects; D005110:Extracellular Space / Q000187:drug effects / Q000201:enzymology*; D020404:Glycerophospholipids / Q000378:metabolism; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D006868:Hydrolysis / Q000187:drug effects; D008670:Metals / Q000494:pharmacology; D031161:Pseudoalteromonas / Q000187:drug effects / Q000201:enzymology* / Q000302:isolation & purification; D012623:Seawater / Q000382:microbiology*; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D010738:Type C Phospholipases / Q000302:isolation & purification*
D020404:Glycerophospholipids; D008670:Metals; D010738:Type C Phospholipases