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Fig. 4 | BMC Microbiology

Fig. 4

From: Ralstonia solanacearum fatty acid composition is determined by interaction of two 3-ketoacyl-acyl carrier protein reductases encoded on separate replicons

Fig. 4

Function of R. solanacearum FabG1 and FabG2 in fatty acid synthesis reactions. Panel a, Function of R. solanacearum FabG1 and FabG2 in the initial cycle of fatty acid synthesis. The initial cycle of fatty acid synthesis was reconstructed in vitro using a combination of E. coli FabH (EcFabH), KAR (E. coli FabG (EcG) (lane 2), or R. solanacearum FabG1 (RsG1) (lane 3) and FabG2 (RsG2) (lane 4), E. coli FabZ (EcFabZ) and FabI (EcFabI) enzymes, ACP, NADH, and NADPH as cofactors and malonyl-ACP plus acetyl-CoA as substrates to produce butyryl-ACP. To complete the fatty acid synthesis reaction (lanes 5 to 7), E. coli FabB (EcFabB) was added to the reactions (the faint bands are due to the instability of short-chain 3-ketoacyl-ACPs in the electrophoresis gels). Panel b and c, Function of R. solanacearum FabG1 and FabG2 in the fatty acid elongation cycle. The elongation reaction mixture contained E. coli FabB (EcFabB), KAR E. coli FabG (EcG) (panel b, lane 3 or panel c, lane 9) or R. solanacearum FabG1 (RsG1) (panel b, lane 4 or panel c, lane 10), FabG2 (RsG2) (panel b, lane 5 or panel c, lane 11) and E. coli FabZ (EcFabZ), malonyl-ACP plus octanoyl-ACP (panel b) or tetradecanoyl-ACP (panel c) as substrates, and NADH and NADPH as cofactors. RsG1 and RsG2 denote R. solanacearum FabG1 and FabG2, respectively. Lane 1 is the product of octanoyl-ACP and lane 7 is the product of tetradecanoyl-ACP. Lane 6 is the product of 3-hydroxydecanoyl-ACP treated with E. coli FabZ and lane 12 is the product of 3-hydroxyhexadecanoyl-ACP treated with E. coli FabZ

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