Identification and characterisation of human aldose 1-epimerase

Aldose 1-epimerase or mutarotase (EC 5.1.3.3) is a key enzyme of carbohydrate metabolism catalysing the interconversion of the α- and β-anomers of hexose sugars such as glucose and galactose. We identified an open reading frame in the human genome (BC014916) which has high sequence similarity to previously identified bacterial aldose 1-epimerases. This sequence was cloned into a bacterial expression vector, and expressed and purified from this source. Enzyme assays show that the protein has aldose 1-epimerase activity and exhibits a preference for galactose over glucose. Site-directed mutagenesis confirmed the involvement of three residues involved in catalysis and substrate binding.