biostudies-literaturede Almeida SGCUNESP-PROPe Research Office39https://www.ebi.ac.uk/biostudies/studies/S-EPMC8738833biostudies-literatureUnknown12(1)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 <i>Phomopsis stipata</i> 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 <i>P. stipata</i> 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 immobili3 BiotechSaccharification of acid-alkali pretreated sugarcane bagasse using immobilized enzymes from <i>Phomopsis stipata</i>.PMC873883309/2016-PROPedo Santos MGHassanpour MZhang ZGiese ECde Almeida SGCde Mello GFDussan KJda Silva DDVfalseSaccharification of acid-alkali pretreated sugarcane bagasse using immobilized enzymes from <i>Phomopsis stipata</i>.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 <i>Phomopsis stipata</i> 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 <i>P. stipata</i> 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 immobili2022-01-01T00:00:00Z2022 Jan2026-07-09T12:17:00.927Z2025-04-06T09:43:55.587ZS-EPMC87388333507062910.1007/s13205-021-03101-2