The Anabaena genome contains two ORFs that appear to encode glutaminases. The genes were expressed as histidine-tagged fusion proteins in Escherichia coli. The purified proteins possessed glutaminase activity using l-glutamine as the substrate, but differed in biochemical properties. All2934 showed an optimal activity at 20 degrees C and pH 6.0, with a higher affinity for l-glutamine than All4774, which had optimal activity at 37 degrees C and pH 7.5. Remarkably, the glutaminase activity of All2934 was phosphate dependent, while All4774 was phosphate independent. The expression of all2934 and all4774 was analyzed using semi-quantitative reverse transcriptase-PCR. The expression level of all2934 was much higher than that of all4774 under normal and nitrogen-depletion conditions, indicating that All2934 may play an important role in metabolizing glutamine in Anabaena.