Reverse Micellar Extraction of Cellulases with Dioctyl Sodium Sulfosuccinate (AOT) in Isooctane

Biofuels will likely become very important in the future as our sources of fossil fuels become depleted. Lignocellulose is a promising alternative energy source. Specially engineered cellulases from the fungi Trichoderma Reesei are utilized to break down lignocellulose into glucose, which can then be transformed into several different kinds of biofuels. These enzymes are usually discarded with the reaction mixture once there is a large buildup of waste products. This is very wasteful, uneconomical and has negative effects on the environment. By finding a way to separate the enzymes from the reaction mixture without significantly decreasing the enzymatic activity, it should be possible to effectively recycle the cellulases. In the past decades, the protein extraction and purification capabilities of reverse micelles have received much attention due to their many advantages over other methods of downstream processing. For instance, reverse micellar extraction is relatively inexpensive, easy to scale-up, and has the ability to operate continuously. Our aim is to optimize the conditions of a reverse micellar extraction system composed of dioctyl sodium sulfosuccinate (AOT) and isooctane for the separation of cellulases from reaction mixtures in order to recycle the enzymes. This has both economical and environmental advantages. By altering the pH, ionic strength, concentrations of surfactants, and types of salt/surfactants, we should be able to optimize the conditions for a high recovery of enzymatic activity.