This is the first study showing the successful application of waste house wood (WHW) to the pilot-scale production of bioethanol by hydrolysis using diluted acid and fermentation using the ethanologenic recombinant Escherichia coli KO11. The major sugars in the WHW hydrolysate were glucose, mannose and xylose; the percentages were approximately 35%, 35% and 20% (w/w), respectively. In anaerobic fermentation using a 5- l reactor in which the oxygen transfer rate (OTR) was 0 mmol/( l·h), KO11 consumed only 25% of the xylose in the WHW hydrolysate over the examined fermentation time of 100 h; however, hexoses such as glucose and mannose were consumed completely. Microaeration at an OTR of 4 mmol/( l·h) enhanced the xylose utilization ratio of KO11 to 100%, at which the ethanol concentration was 35.4 g/ l and the ethanol yield was 0.42, although the maximum ethanol concentrations were 28.8 and 26.6 g/ l at OTRs of 0 mmol/( l·h) and 15 mmol/( l·h), respectively. Moreover, this microaerobic fermentation at OTR of 4 mmol/( l·h) was applied to 1000- l scale bioethanol production using the WHW hydrolysate. The xylose utilization ratio reached 100% and the ethanol yield was determined to be 0.45 for a 63-h fermentation, which were comparable to those obtained from the laboratory-scale fermentation.