Abstract
We checked the presence of phospholipase $A_2(PLA)_2$ which could split the ester bond at the position 2 in the glycerol backbone of glycerophospholipids, in the cells of hyperthermophiles of Pyrococcus horikoshii and Sulfolobus acidocaldarius. The results obtained are as follows; (1). Pyrococcus horikoshii cells were grown in obligate anaerobic conditions at $95^{\circ}C$ and they needed sulfur as energy source instead of oxygen, while Sulfolobus acidocaldarius species grew well in the aerobic medium (pH 2.5) containing yeast and sucrose at $75^{\circ}C$. (2). Pyrococcus horikoshii cells produced phospholipase $A_2$ in the cell culture media although this species did not show lipase activity at least in the pH range of 1.5 ${\sim}$ 3.5. Sulfolobus acidocaldarius cells produced lipase hydrolyzing triacylglycerols such as triolein, but did not split any kind of phospholipids used as substates. (3). The compound of 1-decanoyl-2-(p-nitrophenylglutaryl) phosphatidylcholine was not suitable for a substrate in this experiment, though frequently used as a subtrate for checking presence of phospholipase $A_2$, for its decomposi-tion in this experiment. The L-${\alpha}$-phosphatidylcholine-${\beta}$-[N-7-nitrobenz-2-oxa-1, 3-diazol]aminohexanoyl-${\gamma}$-hexadecanoyl labelled with a fluorescent material, did not show any migration of acyl chains in the molecule during the reaction with phospholipase $A_2$ under a hot condition. (4). Phospholipase $A_2$ in the cells of Pyrococcus horikoshii, showed the optimum activity at $pH6.7{\sim}7.2$ and $95{\sim}105^{\circ}C$, respectively, and was activated by addition of calcium chloride solution. Andthe phospholipase $A_2$ specifically hydrolyzed glycero-phospholipids such as phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine and phosphatidyl inositol, but could not split phospholipid containing ether bonds in the molecule such as DL -${\alpha}$-phosphatidylcholine-${\beta}$-palmitoyl-${\gamma}$-O-hexadecyl, DL-${\alpha}$-phosphati- dylcholine-${\beta}$- oleoyl-${\gamma}$-O-hexadecyl, DL-phosphatidylcholine-dihexadecyl.