<P><B>Highlights</B></P><P>► Low temperature alkali pretreatment for ethanol production from sugarcane stalks. ► LTA pretreatment greatly enhanced the enzymatic digestibility of sugarcane bagasse. ► Level of delignification was taken as key indicator for pretreatment effectiveness. ► Joint conversion of juice and bagasse to ethanol was accomplished on kilogram scale.</P> <P><B>Abstract</B></P><P>An alternative route for bio-ethanol production from sugarcane stalks (juice and bagasse) featuring a previously reported low temperature alkali pretreatment method was evaluated. Test-tube scale pretreatment-saccharification experiments were carried out to determine optimal LTA pretreatment conditions for sugarcane bagasse with regard to the efficiency of enzymatic hydrolysis of the cellulose. Free fermentable sugars and bagasse recovered from 2kg of sugarcane stalks were jointly converted into ethanol via separate enzymatic hydrolysis and fermentation (SHF). Results showed that 98% of the cellulose present in the optimally pretreated bagasse was hydrolyzed into glucose after 72-h enzymatic saccharification using commercially available cellulase and <I>β</I>-glucosidase preparations at relatively low enzyme loading. The fermentable sugars in the mixture of the sugar juice and the bagasse hydrolysate were readily converted into 193.5mL of ethanol by <I>Saccharomyces cerevisiae</I> within 12h, achieving 88% of the theoretical yield from the sugars and cellulose.</P>