Loosenin treatment of Agave tequilana bagasse. Washed A. tequilana bagasse was cut into pieces and then incubated with 50 mM acetate buffer, 50 mM acetate buffer plus 20 μg of loosenin, or 20 μg of proteins from mock supernatant during 8 h at room temperature. Then, one μl of GC 220 (equivalent to 1 U of endoglucanase) was added and samples taken at the indicated time points to analyze for RS release. Acetate buffer (diamonds); acetate buffer + GC 220 (squares); mock SN [20 μg] + GC 220 (open circles); loosenin [20 μg] (closed circles); loosenin [20 μg] + GC 220 (triangles). All experiments were performed in triplicate, and error bars indicate the standard deviations.