Cloning and expression of fatty acids biosynthesis key enzymes from sunflower (Helianthus annuus L.) in Escherichia coli.
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.
Serrano-Vega
María Josefa
MJ
Instituto de la Grasa, CSIC, Av. Padre García Tejero-4, 41012, Sevilla, Spain.
Venegas-Calerón
Mónica
M
Garcés
Rafael
R
Martínez-Force
Enrique
E
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
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