This study examines the effect of surfactant-enhanced enzymatic hydrolysis on eucalyptus Kraft pulps produced under high (CPHA) and mild (CPMA) alkali conditions to optimize saccharification and sugar yield. Compositional analysis revealed an increase in glucan content, from 40% in untreated eucalyptus to 70.1% in CPHA. Both pulps were hydrolyzed using Cellic^® CTec3 HS enzyme (Novozymes). A 2² factorial design revealed maximum sugar conversion (~100%) with enzyme loading of 10 FPU/g carbohydrate and 10% (w/v) solids. Tween 20 significantly boosted hydrolysis in CPMA, increasing reducing sugars from 42 g/L to 65 g/L and efficiency from 59.6% to 92.2% within 6 h. By contrast, Tween 80 and PEG 400 showed limited effects on CPMA. Surfactants mitigated lignin–enzyme interactions, especially in CPMA, as higher lignin content restricted hydrolysis efficiency. Phenolic content in the hydrolysates revealed that Tween 80 increased the release of inhibitory compounds, while Tween 20 kept phenolic levels lower. Overall, Tween 20 improved sugar yields and hydrolysis efficiency even with moderate lignin removal during kraft pretreatment, highlighting its potential to reduce enzyme loading and costs in industrial biorefineries. This study underscores the importance of optimizing surfactant selection based on biomass composition for effective enzymatic hydrolysis for cellulosic sugar recovery.