Phytoene desaturase is present in a large protein complex in the plastid membrane

Phytoene desaturase (PDS; EC 1.14.99.-) represents one of the key enzymes in the carotenoid biosynthetic pathway and is present in nearly all types of plastids in plants. To further characterize PDS, we isolated the PDS cDNA from cauliflower ( BoPDS ) and confirmed its function by heterologous expression in a strain of Escherichia coli containing a carotenoid-producing plasmid. The BoPDS cDNA encodes a predicted mature protein of approximately 55 kDa. In comparison with PDS from a few other plant species, BoPDS exhibited a high enzyme activity in E. coli , and its expression in plastids was independent of carotenoid levels. Plastids were purified from tissues of different plant species including cauliflower curds, tomato fruits, carrot roots and Arabidopsis leaves. By employing both Blue Native PAGE and SDS-PAGE approaches in conjunction with Western blot analysis, it was found that PDS in these plants existed in two forms. The plastid membrane form was present in a large protein complex of approximately 350 kDa, whereas the stroma version was in an approximately 660 kDa complex.