<p>The present study evaluated the <i>in vitro</i> efficacy of different feed enzymes on degrading non-starch polysaccharide (NSP) residues present in digesta collected from broilers' gastrointestinal tract. Two dietary treatments, wheat-soy or maize-soy based diets, were assigned to broilers (12 replicates per treatment" 10 birds per replicate pen) from d 0 to 35. At 35 d of age, digesta was collected from the gizzard, jejunum and ileum from three birds per replicate and pooled. The digesta samples were digested <i>in vitro</i> with one of nine enzymes" GH10, GH11 and GH8 xylanase, arabinofuranosidase, a combination of GH10 xylanase and β-glucanase (XYN+BG), β-mannanase, protease, cellulase cocktail and pectate lyase. When using the gizzard digesta as substrate, GH8 xylanase reduced (P<0.005) both insoluble and soluble NSP levels compared to the control, regardless of diet type. Analysis of NSP degradation in jejunal digesta from birds fed the wheat-soy diet revealed that the three xylanases and arabinofuranosidase reduced (P<0.001) insoluble NSP level compared to the control, but the soluble NSP level was not affected by the enzyme treatments. All enzymes, except the cellulase cocktail and pectate lyase, reduced (P=0.005) the insoluble NSP level in the jejunal digesta collected from birds fed the maize-soy diet. Similarly, all enzyme preparations reduced (P<0.001) the insoluble NSP level in ileal digesta obtained from birds fed the maize-soy diet. The GH10 and GH11 xylanase, XYN+BG, cellulase cocktail and pectate lyase reduced (P<0.001) the insoluble NSP level in the ileal digesta from birds fed the wheat-soy diet compared to the control, with XYN+BG exhibiting the greatest reduction. Collectively, the present <i>in vitro</i> study indicated that the <i>in vitro</i> efficacy of enzyme preparations was dictated by the amount and type of NSP remaining undigested in various parts of the gut, which perhaps affect the <i>in vivo</i> efficacy.</p>