At pH 4.8 and 50 degrees C, semi-continuous countercurrent saccharification was performed using alpha-cellulose. Every second day, solids and liquids were transferred countercurrently and liquid samples were taken to determine sugar concentration in each stage. Train 1 (5 mg protein/g biomass, 8 stages) achieved 87.8% glucose conversion and 102 g/L total sugar. Train 2 (2 mg/g, 8 stages) achieved 56.1% glucose conversion and 65 g/L total sugar. Train 3 (2 mg/g, 16 bottles) achieved 73.6% glucose conversion and 67 g/L total sugar. Compared to 5-day batch saccharification, to achieve the same glucose conversion, enzyme requirements were reduced by factors of 16.8, 8, and 20.5 for Trains 1, 2, and 3, respectively. Further improvements can be realized by changing the enzyme addition point and increasing the number of stages. (C) 2016 Elsevier Ltd. All rights reserved.