Fatty acyl-AMP as an intermediate in fatty acid reduction to aldehyde in luminescent bacteria

Abstract

The acyl protein synthetase component (50K) of the fatty acid reductase complex from the luminescent system of Photobacterium phosphoreum has been found to catalyze the activation of fatty acid via formation of an enzyme bound acyl-AMP (carboxyphosphate mixed anhydride) immediately prior to the acylation of the enzyme. PPi-ATP exchange and nucleotide binding experiments are dependent on fatty acid and indicate that the fatty acyl-AMP is directly formed and that an adenylated enzyme intermediate is not part of the mechanism. The formation of acyl-AMP from fatty acid and ATP is reversible with a standard free energy of -2 kcal/mol, and is dependent on Mg2+. The fatty acyl-AMP intermediate has been isolated and shown to be part of the pathway of fatty acid reduction. The 34K component of the complex, which strongly stimulates the acylation of the 50K protein by fatty acyl-AMP or fatty acid and ATP, is not required for the formation of acyl-AMP showing that it differentially affects the fatty acid activation and acylation steps catalyzed by the 50K protein.