Permeabilization of Aspergillus niger by Reverse Micellar Solutions and Simultaneous Purification of Catalase

Abstract

Reverse micellar solutions(RMS) of sodium bis‐ (2‐ethylhexyl)‐sulfosuccinate (AOT) in aliphatic organic solvents were used for permeabilization and protein removal directly from Aspergillus niger cells. Most of the cell wall proteins (∼95–100%) were solubilized into the reverse micelles solutions. A significant fraction of intracellular catalase (26–30%) permeates out of the cells and remains on the cell surface and is recovered in a cell wash. Because of it's size the catalase in not solubilized in the RM water pools and thus is not detected in the stripping solution used to recover other proteins from the organic reverse micellar solutions. The remaining amount of catalase is recovered by breaking the cells in purer form. Multiple extractions were used for the extraction of the cell wall proteins followed by ultrasonication of the cells to recover intracellular catalase in a purified form. Therefore, catalase with 5 fold purification was recovered in 88% yield from the RMS‐ treated cells.