title: Biochemical characterization of a delta12 acyl-lipid desaturase after overexpression of the enzyme in Escherichia coli.
abstract: The Delta12 acyl-lipid desaturase of Synechocystis sp. PCC 6803 was overexpressed in Escherichia coli as an active enzyme. The overexpressed protein was associated with cell membranes; it represented about 10% of the total cellular protein and 25% of the total membrane protein. The enzyme in the membrane fraction exhibited strong fatty-acid desaturase activity. The desaturase in salt-washed membranes was stabilized by the presence of sorbitol. Storage of salt-washed membranes in 2 M sorbitol at 4 degrees C and at pH 7-8 for six days resulted in the loss of less than 10% of the desaturase activity. The desaturase activity had a positive temperature coefficient, a result that suggests that the increase in the desaturation of fatty acids at low temperature might not be caused by the activation of desaturases at low temperature but, rather, by the increased synthesis of desaturases de novo.
authors.affiliation: Department of Molecular Biomechanics, the Graduate University for Advanced Studies, Myodaiji, Okazaki 444, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Panpoom
authors.forename: D A
authors.initials: DA
authors.lastname: Los
authors.forename: N
authors.initials: N
authors.lastname: Murata
mesh_terms: D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics; D004790:Enzyme Induction; D004795:Enzyme Stability; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000096:biosynthesis / Q000378:metabolism*; D015870:Gene Expression; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D010957:Plasmids; D013696:Temperature
chemical_list: D007544:Isopropyl Thiogalactoside; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase