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Biotin is an essential cofactor required for carboxylation and decarboxylation reactions in all domains of life. While biotin biosynthesis in most Bacteria and Eukarya is well studied, the complete pathway for this vitamer in Archaea is still not known. Detailed genome searches indicated the presence of possible  bio  gene clusters only in Methanococcales and Thaumarchaeota. Therefore, we analysed the functionality of the predicted genes  bioA ,  bioB ,  bioD  and  bioF  in the Thaumarchaeon  Nitrososphaera gargensis  Ga2.9 which are essential for the later steps of biotin synthesis. In complementation tests, the gene cluster-encoded  N .  gargensis bioABD  genes except  bioF  restored growth of corresponding  E .  coli  Rosetta-gami 2 (DE3) deletion mutants. To find out how biotin biosynthesis is initiated, we searched the genome for a possible  bioH  analogue encoding a pimeloyl-ACP-methylester carboxylesterase. The respective amino acid sequence of the ORF  estN1  showed weak conserved domain similarity to this class of enzymes (e-value 3.70e −42 ). Remarkably, EstN1 is a promiscuous carboxylesterase that complements  E .  coli ΔbioH  and  Mesorhizobium loti ΔbioZ  mutants for growth on biotin-free minimal medium. Additional 3D-structural models support the hypothesis that EstN1 is a BioH analogue. Thus, this is the first report providing experimental evidence that Archaea carry functional  bio  genes.

The Thaumarchaeon N. gargensis carries functional bioABD genes and has a promiscuous E. coli ΔbioH-complementing esterase EstN1