Dedicated to Prof. Dr. André Pirson on the occasion of his 90th birthday.
ISOLATION AND CHARACTERIZATION OF PARAMYLON SYNTHASE FROM EUGLENA GRACILIS (EUGLENOPHYCEAE)1,
Article first published online: 21 DEC 2001
Journal of Phycology
Volume 37, Issue 1, pages 38–46, February 2001
How to Cite
Bäumer, D., Preisfeld, A. and Ruppel, H. G. (2001), ISOLATION AND CHARACTERIZATION OF PARAMYLON SYNTHASE FROM EUGLENA GRACILIS (EUGLENOPHYCEAE)1,. Journal of Phycology, 37: 38–46. doi: 10.1046/j.1529-8817.2001.037001038.x
- Issue published online: 21 DEC 2001
- Article first published online: 21 DEC 2001
- 1 Received 8 June 2000. Accepted 26 September 2000.
- Euglena gracilis;
- paramylon synthesis;
- photoaffinity labeling
The aim of this study was to isolate and characterize the paramylon synthesizing enzyme from Euglena gracilis Klebs. A method for enzyme solubilization with high synthase activity using the zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate is presented. Fractionated purification showed that the main enzyme activity was associated with the paramylon granula fraction, isolated from heterotrophically grown cells of E. gracilis. Further purification by sucrose density centrifugation resulted in a large enzyme complex with an apparent molar mass of 670 kDa (native). The complex remained active throughout the isolation procedures and produced beta-1,3-glucan in vitro. Two polypeptides of 37 and 54 kDa could be identified by photoaffinity labeling with [32P]-UDP-glucose as substrate after SDS-PAGE.