An efficient conversion of lignocellulose into fermentable sugars is a key step in producing bioethanol in a cost effective and eco-friendly manner. Alternative source like water hyacinth biomass (WHB) (Eichhornia crassipes) may be used as a supplement for the routine feedstocks. The enzyme loading for optimum yield of total reducing sugar was investigated and the enzyme-substrate interaction optimised. The maximal reducing sugar and xylose yield was obtained using cellulase and xylanase loading of 46.12 and 289.98 U/g and 2.26% (w/v) substrate loading. The efficiencies of ethanol production from the WHB hydrolysate are very less and the maximal ethanol yield was 3.4969 g/L when Pichia stiptis was used, followed by 3.4496 and 3.1349 g/L for Candida shehatae and Saccharomyces cerevisiae.

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