The one-pot production of a promising biofuel, 5-ethoxymethylfurfural (EMF) from corn stover with mixed acid as the catalyst, was investigated and optimized by using the response surface methodology. Catalyzed by the mixed acids comprised of 0.1% H2SO4 and 1.0% zeolite USY, a high EMF yield of 23.9% at 210 degrees C and 125 min can be obtained from corn stover. Meanwhile, the catalytic activity of the zeolite USY had no obvious loss after five recycles. On the basis of the putative reaction pathway, the kinetics modeling for the EMF production directly from corn stover was developed, which coincided with the first-order reaction model. This study provides a promising strategy for EMF preparation from corn stover.