EXTRACTION OF CHARGED FUSION PROTEINS IN REVERSED MICELLES - COMPARISON BETWEEN DIFFERENT SURFACTANT SYSTEMS

Behavior of a series of fusion proteins of varying charge in reversed micellar extraction was studied. The proteins consisted of the enzyme glucoamylase from Aspergillus awamori joined to short peptides contain ing from 0-10 additional aspartate residues. The fusions were partitio ned into two different cationic surfactant systems, one based on the s urfactant trioctylmethylammonium chloride (TOMAC) and the other on cet yltrimethylammonium bromide (CTAB). These two systems differed chiefly in micelle size as measured by the water to surfactant ratio, w(o). W ater numbers were determined for the TOMAC system, with values of appr oximately 10, and as a function of pH and ionic strength for CTAB for each of the mutant enzymes. For the CTAB system, water numbers were as low as 50 with NaCl concentrations of 500 mM and as high as 68 at 300 mM NaCl (95% confidence level of 2.4). The enzyme partitioned most st rongly using CTAB, with maximal. recoveries' approaching 95%. However, in the CTAB system, there were no significant differences in behavior between the mutants because of the relatively large micellar size, ev en under high salt concentrations. Extraction of the control enzyme fr om clarified cell broth indicated that broth components did not signif icantly interfere with partitioning.