Saccharification of acid–alkali pretreated sugarcane bagasse using immobilized enzymes from Phomopsis stipata

In this study, a mild-temperature two-step dilute acid and alkaline pretreatment (DA-AL) process was developed to generate highly digestible cellulose pulp from sugarcane bagasse for producing fermentable sugars by novel thermophilic cellulases derived from Phomopsis stipata SC 04. First, DA pretreatment of sugarcane bagasse at 2% (w/v) H2SO4 and 121 °C for 71 min, followed by AL pretreatment at 2.2% (w/v) NaOH and 110 °C for 100 min led to the pulp containing 86% cellulose. The cellulose pulp was hydrolyzed by the immobilized P. stipata cellulase on Ca-alginate beads, following optimization of immobilization conditions. The results showed that mixing the cellulase extract and sodium alginate solutions at a volume ratio of 1:4 led to the highest immobilization efficiencies of 99.83% for β-glucosidase and 97.52% for endoglucanase while the enzyme leakage was the lowest. The use of the immobilized cellulases led to a cellulose digestibility of 30% in the initial batch and recycling of the immobilized cellulases reduced cellulose digestibility to 18% after s recycling for two times (a total of third rounds). Overall, this study provides useful information in the use of a mild pretreatment process to produce highly digestible cellulose pulp and in the immobilization of thermophilic cellulases to produce fermentable sugars from pretreated biomass.