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

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The native pyrenoid core matrix of the green alga Bryopsis maxima was isolated by diethyl ether treatment and sucrose density gradient centrifugation using 1.8 M phosphate buffer. The purity of the pyrenoids was examined by microscopy, polyacrylamide gel electrophoresis and marker materials. The purified pyrenoids contained the large subunit and the small subunit of ribulose 1,5-bisphosphate carboxylase (RuBPCase) and more than 10 minor polypeptides. They also showed RuBPCase activity when solubilized on being transferred to a low-concentration buffer. The specific activity was 0.62μmol CO_2 fixed・(mg protein)^<-1>・min^<-1>. This isolation method is suitable for obtaining intact pyrenoids not covered by starch sheaths or membranes without the need for chloroplast fixation.
日本植物生理学会の論文