Isolation of Native Pyrenoid Core Matrix Having Ribulose 1,5-bisphosphate Caxrboxylase Activity from the Green Alga Bryopsis maxima

The native pyrenoid core matrix of the green alga Bryopsis maximawas isolated by diethyl ether treatment and sucrose densitygradient centrifugation using 1.8 M phosphate buffer. The purityof the pyrenoids was examined by microscopy, polyacrylamidegel electrophoresis and marker materials. The purified pyrenoidscontained the large subunit and the small subunit of ribulose1,5-bisphosphate carboxylase (RuBPCase) and more than 10 minorpolypeptides. They also showed RuBPCase activity when solubilizedon being transferred to a low-concentration buffer. The specificactivity was 0.62 µmol CO₂ fixed (mg protein)^–1min^–1. This isolation method is suitable for obtainingintact pyrenoids not covered by starch sheaths or membraneswithout the need for chloroplast fixation. (Received July 27, 1987; Accepted October 20, 1987)