Two aerobic pathways for the formation of unsaturated fatty acids in Pseudomonas aeruginosa.
The double bond in anaerobic unsaturated fatty acid (UFA) biosynthesis is introduced by the FabA dehydratase/isomerase of the bacterial type II fatty acid biosynthetic pathway. A DeltafabA mutant of Pseudomonas aeruginosa grew aerobically, but required a UFA supplement for anaerobic growth. Wild-type cells produced 18:1Delta11 as the principal UFA, whereas the DeltafabA strain produced only 16:1Delta9. The double bond in the 16:1Delta9 was introduced after phospholipid formation and was localized in the sn-2 position. Two predicted membrane proteins, DesA and DesB, possessed the conserved histidine clusters characteristic of fatty acid desaturases. The DeltafabADeltadesA double mutant required exogenous fatty acids for growth but the DeltafabAdesB double mutant did not. Exogenous stearate was converted to 18:1Delta9 and supported the growth of DeltafabADeltadesA double mutant. A DeltafabADeltadesAdesB triple mutant was unable to desaturate exogenous stearate and was an UFA auxotroph. We detected a 2.5-fold increase in desA expression in DeltafabA mutants, whereas desB expression was derepressed by the deletion of the gene encoding a transcriptional repressor DesT. These data add two aerobic desaturases to the enzymes used for fatty acid metabolism in proteobacteria: DesA, a 2-position phospholipid Delta9-desaturase that supplements the anaerobic FabA pathway, and DesB, an inducible acyl-CoA Delta9-desaturase whose expression is repressed by DesT.
Zhu
Kun
K
Department of Infectious Diseases, Protein Science Division, St Jude Children's Research Hospital, Memphis, TN 38105, USA.
Choi
Kyoung-Hee
KH
Schweizer
Herbert P
HP
Rock
Charles O
CO
Zhang
Yong-Mei
YM
D000332:Aerobiosis / Q000235:genetics; D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000201:enzymology; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis* / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006836:Hydro-Lyases / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D013228:Stearates / Q000378:metabolism; D013230:Stearoyl-CoA Desaturase / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
D005231:Fatty Acids, Unsaturated; D008565:Membrane Proteins; D013228:Stearates; D013230:Stearoyl-CoA Desaturase; D006836:Hydro-Lyases