Utilization of a protein‐based fat substitute (100% replacement) in a muffin with and without added fungal protease, fungal amylase and/or bacterial amylase was compared with a full fat counterpart. The enzymes were evaluated independently and in combination with each other. Physical tests indicated no significant differences (P>0.05) among any of the variations in volume, crumb L and crust L and b values. The full fat muffin was significantly (P<0.05) more tender, lower in moisture, and more yellow of crumb when compared with the other variations. The 100% fat substituted muffin with enzymes, generally, had a lower moisture content, lower volume, decreased staling rate, and decreased crumb firmness when compared with the 100% fat substituted muffin without enzymes.