<P><B>Abstract</B></P> <P>Using lignocellulosic residues for bioethanol production could provide an alternative solution to current approaches at competitive costs once challenges related to substrate recalcitrance, process complexity and limited knowledge are overcome. Thus, the impact of different process variables on the ethanol production by <I>Saccharomyces cerevisiae</I> using the hemicellulosic fraction extracted through the steam-treatment of softwood chips mixed with construction and demolition residues was assessed. A statistical design of experiments approach was developed and implemented in order to identify the influencing factors (various nutrient addition sources as well as yeast inoculum growth conditions and inoculation strategies) relevant for enhancing the ethanol production potential and substrate uptake. Ethanol yields of 74.12% and monomeric sugar uptakes of 82.12 g/L were predicted and experimentally confirmed in bench and bioreactor systems. This innovative approach revealed the factors impacting the ethanol yields and carbohydrate consumption allowing powerful behavioral predictions spanning different process inputs and outputs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> 7 influencing factors concerning yeast inoculum and nutrient sources were screened. </LI> <LI> Yeast inoculum variables as well as backset addition were the most significant. </LI> <LI> Several different optimization scenarios were developed. </LI> <LI> A maximum of 81.24% of hemicellulosic ethanol yield was predicted. </LI> <LI> Optimization confirmations and bioreactor scale-up were performed. </LI> </UL> </P>