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Heme is an essential prosthetic group for many proteins involved in fundamental biological processes in all three domains of life. In  Eukaryota  and  Bacteria  heme is formed  via  a conserved and well-studied biosynthetic pathway. Surprisingly, in  Archaea  heme biosynthesis proceeds  via  an alternative route which is poorly understood. In order to formulate a working hypothesis for this novel pathway, we searched 59 completely sequenced archaeal genomes for the presence of gene clusters consisting of established heme biosynthetic genes and colocalized conserved candidate genes. Within the majority of archaeal genomes it was possible to identify such heme biosynthesis gene clusters. From this analysis we have been able to identify several novel heme biosynthesis genes that are restricted to archaea. Intriguingly, several of the encoded proteins display similarity to enzymes involved in heme  d   1  biosynthesis. To initiate an experimental verification of our proposals two  Methanosarcina barkeri  proteins predicted to catalyze the initial steps of archaeal heme biosynthesis were recombinantly produced, purified, and their predicted enzymatic functions verified.

A Novel Pathway for the Biosynthesis of Heme inArchaea: Genome-Based Bioinformatic Predictions and Experimental Evidence