Cloning and expression of a lipoxygenase from Pseudomonas aeruginosa 42A2.
In order to produce (S) 10-monohydroxy-8E-octadecenoic acid (MHOD) from oleic acid, a full-length probable lipoxygenase cDNA from Pseudomonas aeruginosa 42A2 was cloned and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by affinity chromatography to electrophoretic homogeneity and specifically stained. Its molecular mass was 70 kDa. The activity of the rec-LOX with oleic acid was about 30% of that of the preferred substrate, linoleic acid (100%). Bacterial LOX forms a new subfamily in the lipoxygenase phylogenetic tree.
Vidal-Mas
Jaume
J
Laboratori de Microbiologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona E-08028, Spain.
Busquets
Montserrat
M
Manresa
Angeles
A
D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019787:Linoleic Acid / Q000378:metabolism; D008084:Lipoxygenase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019301:Oleic Acid / Q000378:metabolism; D010802:Phylogeny; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D019301:Oleic Acid; D019787:Linoleic Acid; D008084:Lipoxygenase