In this work, a valorization of the starch stemming from downgraded potatoes was approached through the preparation of starch nanoparticles using different physical methods, namely liquid and supercritical carbon dioxide, high energy ball milling (HEBM), and ultrasonication on the one hand and enzymatic hydrolysis on the other hand. Starch nanoparticles are beneficial as a reinforcement in food packaging technology as they enhance the mechanical and water vapor resistance of polymers. Also, starch nanoparticles are appropriate for medical applications as carriers for the delivery of bioactive or therapeutic agents. The obtained materials were characterized using X-ray diffraction as well as scanning and transmission electron microscopies (SEM and TEM), whereas the hydrolysates were analyzed using size exclusion chromatography coupled with pulsed amperometric detection (SEC-PAD). The acquired results revealed that the physical modification methods led to moderate alterations of the potato starch granules' size and crystallinity. However, enzymatic hydrolysis conducted using Pullulanase enzyme followed by nanoprecipitation of the hydrolysates allowed us to obtain very tiny starch nanoparticles sized between 20 and 50 nm, much smaller than the native starch granules, which have an average size of 10 μm. The effects of enzyme concentration, temperature, and reaction medium pH on the extent of hydrolysis in terms of the polymer carbohydrates' fractions were investigated. The most promising results were obtained with a Pullulanase enzyme concentration of 160 npun/g of starch, at a temperature of 60 °C in a pH 4 phosphate buffer solution resulting in the production of hydrolysates containing starch polymers with low molecular weights corresponding mainly to P-10, P-5, and fractions with molecular weights lower than P-5 Pullulan standards.
Keywords: Pullulanase; enzymatic hydrolysis; high-energy ball milling; potato starch; supercritical CO2; ultrasonication.