Optimization of cost-effective enzymatic saccharification using low-cost protic ionic liquid as pretreatment agent in Agave bagasse

Abstract

This work studied the optimization of enzymatic saccharification of Agave tequilana bagasse (ATB) pretreated with the low-cost protic ionic liquid (PIL) ethanolamine acetate ([EOA][OAc]) using the highly available and cost-effective mixture of the enzymatic cocktails Celluclast 1.5L-Viscozyme L. Response surface methodology (RSM) was employed to maximize the sugars concentration and yield. The RSM optimization conditions of the enzymatic saccharification of pretreated ATB that achieved the maximum reducing sugars (RS) concentration were: 11.50 % w/v solids loading, 4.26 pH with 0.76 and 1.86 mg protein/mL buffer of Viscozyme L and Celluclast 1.5L, respectively. Similarly, the conditions that maximize the sugar yield (SY) were solids loading of 5.62 % w/v, and 4.51 pH as well as 1.07 and 2.03 mg protein/mL buffer of Viscozyme L and Celluclast 1.5L, respectively. Saccharification performance of the first-generation and low-cost enzyme mixture Celluclast 1.5L-Viscozyme L was compared with that reached by a second-generation and higher-cost CTec2, where Celluclast 1.5L-Viscozyme L achieved 60.86 ± 2.66 % y 79.25 ± 3.34 % of the sugars released by CTec2 at the same hydrolysis time (12 h) for the sugar concentration and yield models, respectively. These results are encouraging since they positively contribute to cost reduction and availability issues, which are key parameters to consider when thinking about scaling-up the process.

Keywords: Agave bagasse; Optimization; Pretreatment; Protic ionic liquid; Saccharification.