Enzymolysis of chitosan by papain and its kinetics

• Optimization of the enzymolysis process using response surface methodology.
• Enzymolysis obeyed the pseudo-second-order and Haldane kinetic model.
• Substrate inhibition was a rate limiting step in the enzymolysis.

Low molecular weight chitosan (LMWC) was obtained by the enzymolysis of chitosan by papain. Enzymolysis conditions (initial chitosan concentration, temperature, pH and ratio of papain to chitosan) were optimized by conducting experiments at three different levels using the response surface methodology (RSM) to obtain high soluble reducing sugars (SRSs) concentrations. Meanwhile, the influence of chitosan substrate concentration on the activity of papain was assessed in the experiments. The enzymolysis process was analyzed using pseudo-first-order and pseudo-second-order kinetic models and the experiment data were found to be more consistent with the pseudo-second-order kinetic model. In addition, the kinetic behavior of the enzymolysis was also investigated by using Haldane model, and chitosan exhibited substrate inhibition. It was clear that the Haldane kinetic model adequately described the dynamic behavior of the chitosan enzymolysis by papain. When the initial chitosan concentration was above 8.0 g/L, the papain was overloaded and exhibited significant inhibition.