Purification and characterization of oxygen-inducible haem catalase from oxygen-tolerant Bifidobacterium asteroides

Bifidobacterium asteroides, originally isolated from honeybee intestine, was found to grow under 20% O-2 conditions in liquid shaking culture using MRS broth. Catalase activity was detected only in cells that were exposed to O-2 and grown in medium containing a haem source, and these cells showed higher viability on exposure to H2O2. Passage through multiple column chromatography steps enabled purification of the active protein, which was identified as a homologue of haem catalase on the basis of its N-terminal sequence. The enzyme is a homodimer composed of a subunit with a molecular mass of 55 kDa, and the absorption spectrum shows the typical profile of bacterial haem catalase. A gene encoding haem catalase, which has an amino acid sequence coinciding with the N-terminal amino acid sequence of the purified protein, was found in the draft genome sequence data of B. asteroides. Expression of the katA gene was induced in response to O-2 exposure. The haem catalase from B. asteroides shows about 70-80% identity with those from lactobacilli and other lactic acid bacteria, and no homologues were found in other bifidobacterial genomes.