Muscadine grapes (Vitis rotundifolia) are widely grown in the southern United States, as the more common Vitis vinifera cannot be cultivated due to Pierce's disease. There is interest to determine if certain cultivars can be used for good-quality wine production. This study compared the effect of pectolytic enzyme pretreatment with conventional skin-contact fermentation on Muscadine (Noble, Vitis rotundifolia) wine major volatiles, aroma active volatiles, and volatile sulfur compounds (VSCs). Volatile composition, aroma activity, and VSCs in the initial juice and wine samples after 3 years were determined by gas chromatography in combination with mass spectrometry (GC-MS), olfactory detection (GC-O), and pulsed flame photometric detection (GC-PFPD). Forty-three nonethanol MS volatiles were common to all samples. Total ion chromatogram (TIC) MS peak area increased 91% in the skin-contact wines from the initial juice but only 24% in the enzyme-treated wine. Thirty-one VSCs were detected. Twenty-four sulfur volatiles were identified by matching their retention characteristics on polar and nonpolar columns with those of standards or MS spectrum matches. Six of these (sulfur dioxide, 1-propanethiol, 3-mercapto-2-pentanone, 3-mercapto-2-butanone, 2,8-epithio-cis-p-menthane, and 1-p-menthene-8-thiol) were reported for the first time in muscadine wine. Five additional VSCs were tentatively identified by matching standardized retention values with literature values, and two remain unidentified. Total sulfur peak areas increased 400% in the skin-contact wine and 560% in the enzyme-treated wine compared to the initial juice. There were 42 aroma-active volatiles in the initial juice, 48 in the skin-contact wine, and 66 in the enzyme-treated wine. Eleven aroma-active volatiles in the skin-contact wine and 16 aroma volatiles in the enzyme-treated wine appear to be due to sulfur volatiles. Pectolytic enzyme-treated wines contained less total volatiles but more sulfur and aroma-active volatiles than the traditional skin-contact wine.