Role of two 5-aminolevulinic acid biosynthetic pathways in heme and secondary metabolite biosynthesis in Amycolatopsis orientalis

Analysis of the Amycolatopsis orientalis genome revealed that two genes, hemA1 and hemA2, belonging to divergent pathways, were involved in the biosynthesis of 5-aminolevulinic acid. The roles of hemA1 and hemA2 were elucidated via genetic manipulation and metabolite analysis. The disruption of hemA1, encoding the glutamyl-tRNA^Glu reductase of the C₅ pathway, was essential for cell growth and is used for heme synthesis. Overexpression of hemA1 resulted in elevated vancomycin and ECO-0501 production in Amycolatopsis orientalis, and it was also effective in increasing the production of daptomycin and natamycin in other Streptomycetes. The disruption of hemA2 indicated that it encodes the 5-aminolevulinic acid synthase of the Shemin pathway, serving as a key enzyme for the synthesis of the precursor aminohydroxycyclopentenone unit of ECO-0501. However, hemA2 disruption could not be complemented by the addition of 5-aminolevulinic acid or by the expression of hemA2 outside of the ECO-0501 gene cluster. The synthesis of ECO-0501 was only restored by the insertion of hemA2 at its original locus. The hemA2 gene could partly complement the hemA1 deficiency. Overexpression of hemA1, a key gene from the heme biosynthetic pathway, is proposed here as a new approach to improve the production of secondary metabolites in bacteria, whereas hemA2 plays different roles depending on its pattern of expression.