Cassava starch, typically, has resistant starch type 3 (RS3) content of 2.4%. This paper shows that the RS3
yields can be substantially enhanced by debranching cassava starch using pullulanase followed by high
pressure or cyclic high-pressure annealing. RS3 yield of 41.3% was obtained when annealing was carried
out at 400 MPa/60 C for 15 min, whereas it took nearly 8 h to obtain the same yield under conventional
atmospheric annealing at 60 C. The yield of RS3 could be further significantly increased by annealing
under 400 MPa/60 C pressure for 15 min followed by resting at atmospheric pressure for 3 h 45 min,
and repeating this cycle for up to six times. Microstructural surface analysis of the product under a
scanning electron microscope showed an increasingly rigid density of the crystalline structure formed,
confirming higher RS3 content.