Mechanism of enhancement of prochymosin renaturation by solubilization of inclusion bodies at alkaline pH.

The renaturation efficiency of recombinant prochymosin depends on not only the renaturation conditions but also the solubilization (denaturation) conditions. Compared with pH 8, solubilization of prochymosin-containing inclusion bodies at pH 11 (8 mol/L urea) results in onefold increase of renaturation efficiency ( approximately 40% vs. approximately 20 %). Alkaline pH facilitates the solubilization of inclusion bodies via the breakage of intermolecular disulfide bonds. Moreover, alkaline pH renders prochymosin molecules to be in a more reduced and more unfolded state which undergoes refolding readily.