Which were generated after the use of individual enzymatic pretreatment, were neutralized in the sequential treatment process. Although numerous reports are available on the sequential use of xylanase and laccase for pulp pretreatment, this is the first report on the sequential application of xylanase and laccase for biobleaching of the agro-residual pulp. SA was selected as the mediator because of its plant origin and its higher efficiency to decolorize the Azure-B. Further, the improvement observed in the residual activity of the enzyme in the presence of pulp could be due to the cellulosic content of the pulp. Pulp fibers might BAY 43-9006 abmole bioscience provide an additional substrate for free radicals generated by the action of the natural mediator. The rate at which these free radicals target the enzyme is decreased owing to the presence of the pulp fiber; therefore, the residual enzyme activity increased. A similar observation was also reported by Fillat et al. Use of natural mediators will be beneficial in pollution reduction because the higher concentration of free radicals generated due to the action of synthetic mediators that make the whole bleaching process highly toxic and raise environmental concerns. Since commercial laccase has lower specific activity, it was less effective for delignification compared to C. subvermispora laccase, which has higher specific activity. Improvement in the strength after enzymatic pretreatment is directly related to the hydrolytic action of xylanase. Hydrolytic enzymes disrupt the surface of cellulosic fiber and generate microfibrils, which in turn lead to a cross-networked, condensed packing of pulp fibers and hence they give extra strength. Since only xylanase has hydrolytic properties, even sequential application of oxidizing enzymes did not improve the strength significantly. Although commercial xylanase alone has been used previously, no report is available on the sequential application of xylanase and laccase for processing wheat straw pulp. Improvement in crystallinity means a decrease in amorphous cellulose and increase in crystalline cellulose of the pulp. The increase in crystallinity might be due to the removal of hemicelluloses and lignin and components adhered to lignin as a result of sequential pretreatment, thereby increasing the cellulose content of the pulp. Sequential pretreatment strategies involve separate laccase supplementation, and this is probably the reason for higher increase in crystallinity in sequential approach than in single enzyme strategy. Achieving less or non-toxic discharge from pulp and paper industry is the biggest challenge in the current scenario. Specific characteristics of the effluents such as BOD and color were determined, and strategy II was found to be the most effective in reducing the pollution load, resulting in 25.8% reduction. This might be because of the sequential use of B. stearothermophilus SDX xylanase and C. subvermispora laccase that removes a significant amount of lignin during pretreatment, which is further removed in the subsequent washes.