Life in unusual environments: progress in understanding the structure and function of enzymes from extreme halophilic bacteria.

Extreme halophilic archaea are saturated with salt and the intracellular electrolyte concentration exceeds that of the extracellular environment. Enzymes and other proteins from extreme halophilic archaea have been purified for many years and studied by biochemical and biophysical solution methodologies. They are active and stable at multimolar salt concentrations and denature below 2 to 3 M NaCl or KCl. Adaptation to these high concentrations of salt, genetic and evolutionary aspects, and the possibility of biotechnological applications are problems of considerable interest. Since the status of this fascinating field of research was reviewed in 1992, malate dehydrogenase from Haloarcula marismortui, now known to be a tetramer, was sequenced, its gene was cloned and expressed in active form, and its physical properties were redefined. A single mutation of Arg100 (in the enzyme active site) to Gln switched the enzyme specificity from malate to lactate dehydrogenase. Recent determination of its molecular structure by X-ray crystallography (O. Dym et al., in press) provides an exciting basis for the understanding of the structure and function of extreme halophilic enzymes. A major problem which so far has not been tackled in the study of extreme halophilic archaea is the understanding of protein nucleic acid interactions which are essential for the performance of biological function. Whereas the stability and activity of enzymes and other proteins can be modified to perform at high salt concentrations by use of currently known structural concepts, the existence of meaningful protein nucleic acid interactions in physiological concentrations of 4 to 5 M KCl constitutes an unsolved enigma worth intensive investigation.