Biomaterial Database

Purification and properties of the fatty acid synthetase complex from the marine dinoflagellate, Crypthecodinium cohnii. 1982

U Sonnenborn, and W H Kunau

De novo biosynthesis of fatty acids in the heterotrophic marine dinoflagellate, Crypthecodinium cohnii, has been studied in vitro. Fatty acid synthetase was located in the cytosol and its activity was dependent on acetyl-CoA, malonyl-CoA, NADPH2 and NADH2. The enzyme was purified 100-fold using ion-exchange chromatography on DEAE-Sephadex A-25, adsorption to hydroxyapatite and gel filtration on Sepharose 4B columns. Very active endogenous proteases were separated from the fatty acid synthetase at the first step of purification. The purified enzyme had a molecular weight of about 400000, as judged from gel filtration, sucrose density gradient centrifugation and polyacrylamide gel electrophoresis under non-denaturing conditions. Polyacrylamide gel electrophoresis under denaturing conditions in the presence of SDS and urea revealed one major protein band of Mr 180000, suggesting that the enzyme is composed of two multifunctional subunits of apparently identical molecular weight. Reaction products of the C. cohnii fatty acid synthetase are free fatty acids due to the presence of a thioesterase activity in the purified enzyme complex. The main product is palmitate. Docosahexaenoic acid (C22:6, n-3), the major fatty acid component of C. cohnii lipids, is not directly synthesized by the enzyme.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008970	Molecular Weight	The sum of the weight of all the atoms in a molecule.	Molecular Weights,Weight, Molecular,Weights, Molecular
D003600	Cytosol	Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.	Cytosols
D004141	Dinoflagellida	Flagellate EUKARYOTES, found mainly in the oceans. They are characterized by the presence of transverse and longitudinal flagella which propel the organisms in a rotating manner through the water. Dinoflagellida were formerly members of the class Phytomastigophorea under the old five kingdom paradigm.	Amphidinium,Dinoflagellata,Dinophyceae,Dinophycidae,Dinophyta,Dinophytes,Gambierdiscus toxicus,Gonyaulax,Gymnodinium,Peridinium,Pyrrhophyta,Pyrrophyta,Dinoflagellates
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D013379	Substrate Specificity	A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.	Specificities, Substrate,Specificity, Substrate,Substrate Specificities
D046911	Macromolecular Substances	Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.	Macromolecular Complexes,Macromolecular Compounds,Macromolecular Compounds and Complexes,Complexes, Macromolecular,Compounds, Macromolecular,Substances, Macromolecular
D064429	Fatty Acid Synthases	Enzymes that catalyze the synthesis of FATTY ACIDS from acetyl-CoA and malonyl-CoA derivatives.	Fatty Acid Synthase,Fatty Acid Synthetases,Acid Synthase, Fatty,Acid Synthases, Fatty,Synthase, Fatty Acid,Synthetases, Fatty Acid