Loosenin enrichment, activity and binding to polysaccharides. (a) LOOS1 enrichment. SDS-PAGE of proteins from yeast culture supernatants transformed with empty vector pSAL3 (mock SN), pSAL3-loos1 (loosenin SN), and enriched LOOS1 (loosenin). (b) LOOS1 binds specifically to Avicel (crystalline cellulose) and chitin. The bounded (pellet) and unbounded (SN, supernatant) fractions are shown (top panel). BSA on the other hand, is found in both fractions, indicating unspecific interaction with the matrixes (lower panel). (c) RS release by cellulase from cotton fibers previously incubated with: two different concentrations of loosenin plus cellulase (20 μg black squares, 40 μg gray diamonds); acetate buffer (open triangles); acetate buffer plus cellulase (gray circles); mock SN plus cellulase (open squares); and only LOOS1 (open circles). (d) Specific activity of cotton fibers previously treated with LOOS1, at different pH, plus cellulase. All experiments were performed in triplicate, and error bars indicate the standard deviations.