Insights into the genome of the enteric bacterium Escherichia blattae:: Cobalamin (B12) biosynthesis, B12-dependent reactions, and inactivation of the gene region encoding B12-dependent glycerol dehydratase by a new Mu-like prophage

The enteric bacterium Escherichia blattae has been analyzed for the presence of cobalamin (B-12) biosynthesis and B-12-dependent pathways. Biochemical studies revealed that E blattae synthesizes B-12 de novo aerobically and anaerobically. Genes exhibiting high similarity to all genes of Salmonella enterica serovar Typhimurium, which are involved in the oxygen-independent route of B-12 biosynthesis, were present in the genome of E. blattae DSM 4481. The dha regulon encodes the key enzymes for the anaerobic conversion of glycerol to 1,3-propanediol, including coenzyme B-12-dependent glycerol dehydratase. E. blattae DSM 4481 lacked glycerol dehydratase activity and showed no anaerobic growth with glycerol, but the genome of E. blattae DSM 4481 contained a dha regulon. The E. blattae dha regulon is unusual, since it harbors genes for two types of dihydroxyacetone kinases. The major difference to dha regulons of other enteric bacteria is the inactivation of the dehydratase-encoding gene region by insertion of a 33,339-bp prophage (MuEb). Sequence analysis revealed that MuEb belongs to the Mu family of bacteriophages. The E. blattae strains ATCC 33429 and ATCC 33430 did not contain MuEb. Accordingly, both strains harbored an intact dehydratase-encoding gene region and fermented glycerol. The properties of the glycerol dehydratases and the correlating genes (dhaBCE) of both strains were similar to other B-12-dependent glycerol and diol dehydratases, but both dehydratases exhibited the highest affinity for glycerol of all B-12-dependent dehydratases characterized so far. In addition to the non-functional genes encoding B-12-dependent glycerol dehydratase, the genome of E. blattae DSM 4481 contained the genes for only one other B-12-dependent enzyme, the methylcobalamin-dependent methionine synthase. Copyright (C) 2004 S. Karger AG, Basel.