Horseradish peroxidase delignification of fermented rooibos modifies biomass structural and chemical properties and improves holocellulolytic enzyme cocktail efficacy

Abstract The purpose of the study was to pretreat fermented rooibos biomass with partially purified horseradish peroxidase (HRP) for lignin removal and convert delignified soluble sugars through saccharification a formulated holocellulolytic enzyme cocktail (HEC). HRP extracted from root tissue by membrane filters characterised biochemically. used remove before hydrolysing it HEC. Our findings indicated that is versatile because displayed activity on guaiacol, 8-aminoquinoline, decolourised methylene blue dye. had pH optimum 4.5 mesophilic temperature range. kinetics studies higher affinity towards guaiacol ( K m = 0.082 mg/mL) followed 8-aminoquinoline 0.221 mg/mL). However, catalytic efficiency revealed hydrolysed (63436.48 s − 1 . (59189.81 efficiently. pretreatment significantly removed content increased pores surface as visualised SEM. FTIR validated SEM results showing reductions at 3324.81, 1615.16 1018.75 cm , corresponding crystalline cellulose, holocellulose regions, respectively. pretreated lowest crystallinity index 11.2% compared 20% control. effectively HEC, which released about 10% yield 6% We conclude modified structural chemical properties biomass, making more accessible hydrolytic enzymes.