Fig 3. Calorimetric trace of LMw levan hydrolysis by B. subtilis levansucrase (SacB) in a VP-ITC.

The enzymatic reaction was carried out at 25°C. The reaction took place in the 1.4 mL calorimetric cell, containing 500 nM SacB in the working buffer. The reaction started by the addition of 40 μL of a 30.16 mM LMw levan solution placed in the calorimeter syringe. After this titration the LMw levan concentration in the cell was 0.85 mM. (*) indicates the onset of the “second reaction” For comparison, the inset show a calorimetric trace for a single reaction to completion (for the hydrolysis of L-arginine ethyl ester catalyzed by tripsin from Martínez (2015)).