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The effect of particle size on enzymatic hydrolysis of cassava flour at subgelatinization temperature was investigated. A multiscale physical metrology was developed to study the evolution of different physical-biochemical parameters: rheology, granulometry, and biochemistry. In this study, four fractions of cassava flour based on the particle sizes under 75 µm (CR075), 75–125 µm (CR125), 125–250 µm (CR250), and 250–500 µm (CR500) were screened for enzymatic hydrolysis effect. The results showed that all cassava flour suspensions exhibited a shear-thinning behavior, and the viscosity increased drastically with the increase of particle size. During hydrolysis, the viscosity reduced slightly and the non-Newtonian behavior became negligible beyond 4 h of the process. The particles size for CR075 and CR125 increased steadily in diameter mean. The samples of CR250 and CR500 showed more fluctuation by first decreasing, followed by increasing in particle sizes during the process. The highest hydrolysis yield was found for samples with particle size under 125 µm (89.5–90.7%), suggesting that mechanical treatment of cassava can enhance the bioconversion rate.

Impact of Particle Size on the Rheological Properties and Amylolysis Kinetics of Ungelatinized Cassava Flour Suspensions