Fuel alcohol production yields can be influenced by lignocellulosic biomass loading. High solid loadings (>20 wt%) are suggested to have the potential to produce more products. However, most often, low substrate loadings (<5% solids, w/w) are used to ensure good wetting and enzyme accessibility, and to minimize any inhibitory effect on the hydrolysis products. Here, we analyzed the effect of substrate loading on the enzymatic hydrolysis of hay with non-commercial enzyme products obtained from white-rot fungi. A significant negative effect on hydrolysis was observed when 10 wt% hay loading was used with the commercial enzyme, however, non-commercial enzyme products from white-rot fungi had no impact on hydrolysis in biomass loading rates from 1 to 10 wt%. Moreover, it was estimated that enzymes extracted from white-rot fungi could be used at a concentration of 0.2 FPU/mL at a biomass loading from 1–10 wt%, resulting in 0.17–0.24 g of released reducing carbohydrates per gram of biomass. Higher concentrations did not result in any significant conversion increase. A mixing impact was only observed in test runs at a substrate loading of 10 wt%. The apparently positive features of the non-commercial enzyme mixes give rise to their future use. The combination and upgrade of existing technologies, e.g., efficient pre-treatment, membrane purification, and concentration and efficient product recovery, should result in even higher conversion yields.