Ultrasound effects were investigated on pectinase (PE), xylanase (XLN) and cellulase (CE) activities at different pH, temperatures, and by sonication pre-treatment, comparing the reaction at ultrasound bath (US) and at a mechanical stirring (MS). In general, US increased the activity of the enzymes by 5% for PE, 30% for XLN and 25% for CE compared to MS. US provided a higher activity at extremes pH (pH 3 and 7), mainly for XLN and CE. The substrate and enzyme pre-sonication enhanced the activities. The previous sonication of xylan increased the xylanase activity in almost 30% under US and almost 20% under MS. On the other hand, cellulase pre-sonication increased the activity in 50% under US and 40% under MS. The catalytic efficiency (V-max/K-M) increase 25% for PE and 17% for higher XLN and CE under US. US affected the PE activity at low temperature improving 10% the PE activity, while its effect was more representative at high temperatures, where the enzymatic activities of XLN and CE were 33% and 15% higher. Our results demonstrated that ultrasound can affect enzymes and substrates, making it a powerful tool for enzymatic-catalyzed reactions.