Bacteria-free culture of a colorless euglenoid, Peranema trichophorum, and establishment of a method for flagellar isolation

Abstract

We have recently shown that the anterior flagellum and its mastigonemes play an important role in gliding movement in the colorless euglenoid flagellate, Peranema trichophorum. To facilitate further biochemical analysis of the molecular mechanism of flagellar surface motility, we have developed an improved monoxenic sterile culture method and a method for flagellar isolation. With initial cell densities of Peranema and co-existing food flagellates, Chlorogonium elongatum, at 800 and 30,000 cells/ml, respectively, Peranema reached its maximum cell density of about 10,000 cells/ml in one week. A cold-shock treatment allowed efficient isolation of flagella from the cell body, and light and electron microscope observations confirmed that the isolated flagella retained their intact structure with abundant mastigonemes. SDS-PAGE analysis showed that the preparation contained a predominant protein band of 100 kDa, which was not stained by silver and may therefore be highly glycosylated.