Food industry wastes could be a significant source for value-added biomolecules like proteins, carbohydrates, oil, and polyphenolic compounds. The present work elucidates an application of ultrasound as an intensification tool for the three-phase partitioning (TPP) of anti-diabetic proteins from industrial turmeric waste. Spent turmeric contains 8-12% (w/w) proteins having various therapeutic properties. Maximum protein recovery (66.52%) was attained under optimized TPP conditions such as 60% salt concentration, pH 7, 0.03 g/mL solid loading, 1:1 aqueous to t-butanol phase ratio, and 45 min time. Further, the ultrasound was applied to TPP as a process intensification tool that reduced extraction time from 45 min to 10 min and enhanced the protein recovery by 20%. At 90 W ultrasound power with 40% duty cycle and 22 kHz ultrasonic frequency, maximum recovery was achieved in minimum time requirement. Partial purification by TPP/UA-TPP and molecular weight (14–40 kDa) of proteins was confirmed by SDS-PAGE. The partially purified fractions by UA-TPP exhibited comparable anti-diabetic activity with acarbose standard. The comparison between various extraction techniques showed that UA-TPP leads to 86% protein recovery in just 10 min with the minimum solvent requirement. Thus, UA-TPP promises rapid and enhanced protein recovery from waste turmeric powder which showed anti-diabetic activity.