Method for the hydrolysis of lignocellulosic biomass

A method for increasing sugar yield from a high solids concentration of lignocellulosic biomass and/or for reducing enzyme quantity, the method comprising:

• a. enzymatic hydrolysis of a lignocellulosic biomass to obtain a slurry, wherein the hydrolysis comprises more than two aliquot additions of enzyme and lignocellulosic biomass;
• b. removal of sugars from the slurry to obtain a residual lignocellulosic biomass and washing of the residual lignocellulosic biomass to obtain washed residual lignocellulosic biomass;
• and
• c. hydrolysis, without enzyme addition, of the washed lignocellulosic biomass obtained from step b;
• wherein said sugars comprise glucose:
• xylose at a ratio of 5-8:
• 1 and/or wherein said washed residual lignocellulosic biomass comprises (i) less than about 6% ash and/or (ii) a lignin component comprising at least about 20% intact β-O-4 bonds per 100 C9 units,
• wherein the method does not include any pre-treatment by milling or treatments which require a temperature above 130° C., a pressure above 110 kPa or the use of a strong acid or strong base.