Ammonia treatment of lignocellulose improves carbohydrate degradability, however, low ammonia dose treatment effects and mechanisms are hardly considered. This study describes low dose ammonia treatment of wheat straw in a statistical design of experiments (Taguchi design) to evaluate the effects of ammonia concentration, treatment time and the Solid:Liquid ratio on structure, composition and enzymatic degradability of the residual fractions. The results showed that low ammonia concentration (≤2 w/w % NH3) resulted in a high carbohydrate recovery (>80%) coupled enzymatic hydrolysis of 50% of xylan and 40% of glucan of the treated material using a (hemi-) cellulase enzyme cocktail. This effect coincidences with the relative decrease in ferulic acid by 10% and coumaric acid by more than 50% analysed via pyrolysis-GC–MS, measured as 4-vinyl-phenol and 4-vinyl-guaiacol, respectively. Our findings show that lowering ammonia concentration increased the effect of treatment time on the enzymatic degradability of the residual fraction.