The effective processing method to produce fermentable sugars and modify the microstructure of black soybean okara using bio-ionic liquid (bio-IL) pretreatment and ultrasound-promoted enzymatic hydrolysis was investigated. The morphology and structural characteristics of okara before and after bio-IL pretreatment and enzymatic hydrolysis under different ultrasonic frequencies were analyzed by field emission scanning electron microscope (FE-SEM), X-ray energy dispersive spectrometer (EDS), and Fourier transform infrared spectroscopy (FTIR). Without pretreatment, the production of total reducing sugar (TRS) under ultrasound (40 kHz/300 W) was 3.4 times of that without ultrasound. Using the bio-IL choline acetate ([Ch][OAc]) in water for the pretreatment of black soybean okara, the TRS production of enzymatic hydrolysis was further increased to 5.2 times of that without ultrasound in 4 h of reaction. The analysis by FTIR and EDS showed that the highly structured matrix of okara was unfolded and broken by the action of combining ultrasound and choline acetate pretreatment, due to which the surface structures with large pores were presented to facilitate the reduction of unfavorable hindrance for enzymatic hydrolysis. The simplified kinetic model was proposed to describe the transport and reaction phenomena of enzymes in a solid liquid system by using two kinetic parameters to show the impeded behavior of enzyme within the matrix of okara. The combination of bio-IL pretreatment and ultrasound-promoted enzymatic hydrolysis was able to make the efficient structural changes of black soybean okara to enhance the digestion of enzymes, and the okara could be valorized for use in foods. (C) 2018, The Society for Biotechnology, Japan. All rights reserved.