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ISOLATION AND CHARACTERIZATION OF PARAMYLON SYNTHASE FROM EUGLENA GRACILIS (EUGLENOPHYCEAE) 1,
Author(s) -
Bäumer Dietrich,
Preisfeld Angelika,
Ruppel Hans Georg
Publication year - 2001
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1046/j.1529-8817.2001.037001038.x
Subject(s) - euglena gracilis , biology , biochemistry , enzyme , centrifugation , euglena , atp synthase , chromatography , chloroplast , chemistry , gene
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 [ 32 P]‐UDP‐glucose as substrate after SDS‐PAGE.