Augmented hydrolysis of acid pretreated sugarcane bagasse by PEG 6000 addition: a case study of Cellic CTec2 with recycling and reuse.

In an integrated lignocellulosic biorefinery, the cost associated with the "cellulases" and "longer duration of cellulose hydrolysis" represents the two most important bottlenecks. Thus, to overcome these barriers, the present study aimed towards augmented hydrolysis of acid pretreated sugarcane bagasse within a short span of 16 h using Cellic CTec2 by addition of PEG 6000. Addition of this surfactant not only enhanced glucose release by twofold within stipulated time, but aided in recovery of Cellic CTec2 which was further recycled and reused for second round of saccharification. During first round of hydrolysis, when Cellic CTec2 was loaded at 25 mg protein/g cellulose content, it resulted in 76.24 ± 2.18% saccharification with a protein recovery of 58.4 ± 1.09%. Filtration through 50KDa PES membrane retained ~ 89% protein in 4.5-fold concentrated form and leads to simultaneous fractionation of ~ 70% glucose in the permeate. Later, the saccharification potential of recycled Cellic CTec2 was assessed for the second round of saccharification using two different approaches. Unfortified enzyme effectively hydrolysed 67% cellulose, whereas 72% glucose release was observed with Cellic CTec2 fortified with 25% fresh protein top-up. Incorporating the use of the recycled enzyme in two-stage hydrolysis could effectively reduce the Cellic CTec2 loading from 25 to 16.8 mg protein/g cellulose. Furthermore, 80% ethanol conversion efficiencies were achieved when glucose-rich permeate obtained after the first and second rounds of saccharification were evaluated using Saccharomyces cerevisiae MTCC 180.