The aim of this work was to study the effects of pretreatment process, hydrolysis condition and structural features of lignin on the improving action of surfactants (Tween 20) for enzymatic hydrolysis of pretreated wheat straw, and further to interpret the relation of these factors with the non-productive adsorption of cellulases on lignin. Tween 20 seemed to be more greatly improve cellulose conversion under harsher conditions. The surfactant showed more significant improvement for acid-pretreated substrates than oxidative-pretreated substrates. Highly-condensed lignin and phenolic hydroxyl groups showed much stronger adsorption ability to cellulases, while Tween 20 could well block the lignin-cellulase interactions recovering cellulose hydrolyzability. It was proposed that pretreatments altered lignin structures, resulting in the change of surface properties thus further impacting the lignin-cellulase interactions. Addition of Tween 20 could modify lignin surface properties to change its hydrophobicity, hydrogen bonding ability and surface charges, thus reducing the nonproductive adsorption of proteins.