In this work, conditions for an enzymatic pretreatment prior to NCC isolation from cotton linter were assessed. Different cellulase doses and reaction times were studied within an experimental design and NCC were obtained. At optimal enzymatic conditions (20U, 2 h), a total yield greater than 80% was achieved and the necessary enzymatic treatment time was reduced 90%. Different intensities of enzymatic treatments led to proportional decreases in fiber length and viscosity and also were inversely proportional to the amount of released oligosaccharides. These differences within fibers lead to quantitative differences in NCC: increase in acid hydrolysis yield, reduction of NCC surface charge and crystallinity increase. Benefits produced by enzymatic treatments did not have influence over other NCC characteristics such as their sulfur content (˜1%), size (˜200 nm), zeta potential (˜-50 mV) or degree of polymerization (˜200). Evidence presented in this work would reduce the use of harsh sulfuric acid generating a cleaner stream of profitable oligosaccharides