Hexanoate synthase, a specialized type I fatty acid synthase in aflatoxin B1 biosynthesis

In fungi, fatty acids are biosynthesized by large multifunctional enzyme complexes, the fatty acid synthases (FASs), which catalyze chain assembly in an iterative manner. Many fungal secondary metabolites contain fatty acid moieties, and it is often unclear whether they are recruited from primary metabolism or are biosynthesized de novo by secondary metabolic FASs. The most convincing evidence of such a dedicated FAS comes from the biosyntheses of aflatoxin (AF) and sterigmatocystin (ST) in certain species of the filamentous fungus Aspergillus. Incorporation studies in AF and genetic analyses of ST and AF biosynthesis strongly suggest that their biosyntheses be-in with the production of a C-6 fatty acid by a specialized FAS. The genes encoding the alpha (hexA) and beta (hexB) subunits of this hexanoate synthase (HexS) from the AF pathway in Aspergillus parsiticus SU-1 were cloned and both their gDNAs and cDNAs were sequenced and their transcriptional ends analyzed. Translated amino acid sequences are predicted to result in proteins of 181.3 and 210.5 k-Da, for HexA and HexB, respectively. Comparison of the HexA and HexB sequences with those of the ST FAS subunits and primary metabolic FASs indicated that the secondary metabolic enzymes are members of a well-defined subclass of the FAS family. Phylogenetic predictions and an analysis of GC-bias in AF and ST pathway genes compared with primary metabolic Aspergillus genes were used as a basis to propose a route for the evolution of the AF and ST clusters. (C) 2001 Academic Press.