title: Cloning and expression of fatty acids biosynthesis key enzymes from sunflower (Helianthus annuus L.) in Escherichia coli.
abstract: To further characterize the stearoyl-acyl carrier protein (ACP) desaturase (EC 1.14.99.6) and the acyl-ACP thioesterase FatB (EC 3.1.2.14) activities from sunflower seeds, we cloned, sequenced and expressed the recombinant genes in Escherichia coli. We obtained two partially purified proteins, His-SAD and His-FATB, each of about 45000 Da. The expression of either proteins produced changes in the E. coli fatty acid profile indicating the functionality of the recombinant proteins. While the expression of His-SAD produced an effect similar to that produced by overexpression of the fabA gene, responsible for the fatty acid desaturation in E. coli, the expression of His-FATB gave rise to an unbalance between unsaturated fatty acids and a toxic effect in E. coli.
authors.affiliation: Instituto de la Grasa, CSIC, Av. Padre García Tejero-4, 41012, Sevilla, Spain.
authors.forename: María Josefa
authors.initials: MJ
authors.lastname: Serrano-Vega
authors.forename: Mónica
authors.initials: M
authors.lastname: Venegas-Calerón
authors.forename: Rafael
authors.initials: R
authors.lastname: Garcés
authors.forename: Enrique
authors.initials: E
authors.lastname: Martínez-Force
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005227:Fatty Acids / Q000096:biosynthesis*; D006368:Helianthus / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012639:Seeds / Q000201:enzymology; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D017931:DNA Primers; D005227:Fatty Acids; D011994:Recombinant Proteins; D006899:Mixed Function Oxygenases; C018747:acyl-(acyl-carrier-protein)desaturase; D013869:Thiolester Hydrolases; C018748:oleoyl-(acyl-carrier-protein) hydrolase