Efficient and Scalable Process to Produce Novel and Highly Bioactive Purified Cytosolic Crystals from Bacillus thuringiensis
Jeffrey Chicca, Nicholas R. Cazeault, Florentina Rus, Ambily Abraham, Carli Garceau, Hanchen Li, Samar Magdy Atwa, Kelly E. Flanagan, Ernesto R. Soto, Mary S. Morrison, David Gazzola, Yan Hu, David R. Liu, Martin K. Nielsen, Joseph F. Urban, Gary R. Ostroff, Raffi V. Aroian
Abstract
Bacillus thuringiensis crystal proteins are widely and safely used as insecticides. Recent studies have shown they also can cure gastrointestinal parasitic worm (nematode) infections when ingested. However, reproducible, scalable, and practical techniques for purifying these proteins have been lacking. Here, we address this severe limitation and present scalable and practical methods for large-scale purification of potently bioactive B. thuringiensis crystals and crystal proteins. The resultant product, called purified cytosolic crystals (PCC), is highly compatible with ingestible drug delivery and formulation. Furthermore, there are growing applications in agriculture and insect control where access to large quantities of purified crystal proteins is desirable and where these methods will find great utility.