Albumins are highly conserved proteins that carry out a multitude of physiological functions and exhibit a broad range of catalytic activities. Moreover, amino acid sequence comparisons of the albumin multigene family indicate that albumins diverged from a common ancestor. Here we report that bovine serum albumin (BSA) can catalyze ester hydrolysis at high temperatures (as high as 160 degrees C) well beyond the temperature limits reported for enzymatic catalysis, including for enzymes from known hyperthermophiles. Furthermore, BSA exhibited a similar to 133-fold increase in its turnover number (k(cat)) toward p-nitrophenyl palmitate from 70 to 150 degrees C. When BSA was incubated for 1 h at 150 degrees C in the presence of 25 mM SDS, it retained complete esterase activity, indicating that a catalytically competent orientation of amino acid residues exists in the denatured or partially unfolded protein. However, esterase activity diminished to similar to 50% upon disruption of the protein's disulfide bridges and disappeared completely when BSA was digested by proteases. These results point to a new standard of robustness for biocatalytic activity at high temperatures. Catalytic activity and promiscuity at very high temperatures could have been advantageous to enzymes in primitive organisms evolving in hot environments, making BSA an intriguing model for early enzymes. (c) 2007 Elsevier Inc. All rights reserved.