This expedited award project is a study on protein solubilization characteristics in reversed micelles under conditions where the water pools within the micelles are partially converted to clathrate hydrates. Hydrate formation in reversed micelles may be a novel and viable method to control the solubilization of proteins within the micelles. The present technique to extract proteins from fermentation broths into reversed micelles and subsequently recover them from the micelles involves cycling the ionic strength, promoting solubilization with low ionic strenth, high ionic strength causing desolubilization. Such approaches can weaken enzymes. The extraction approach under investigation is to solubilize protein in reversed micelles at low ionic strength, and ten to induce the formation of clathrate hydrates under relatively low temperatures (273-283 K) and moderate pressures (0.2-5 MPa) to effect protein recovery out of the micelles. Formation of hydrates, which are ice-like structures, would effectively reduce the microaqueous phase volume within the micelles, and thus control desolubilization characteristics. The exploratory research project is divided into three parts (a) the effect of moderately enhanced pressures on protein solubilization in reversed micelles under ambient temperature conditions where hydrate formation is prevented (b) the study of hydrate formation in reversed micelles at low temperatures, and comparisons with hydrate formation in bulk aqueous media, and most importantly, the synthesis of parts (a) and (b), i.e. (c) a study of the effects of hydrate formation on protein solubilization in reversed micelles.
