Hi

please go through these links

https://www.microbiology.biology.upatras.gr/en/protocols/111-determination-of-a-amylase-activity.html

https://www.scielo.br/j/babt/a/NwvTRSHmXNPHFRhZPpb8ZDB/?lang=en

Good luck

Amylase activity can be determined by homogenizing seeds with chilled Tris- HCl buffer or phosphate buffer (pH 6.8), centrifuge at 10,000 g for 10 min at 4°C. Add 1 mL SNT and 1 mL of substrate (0.15% starch, 0.2 mM CaCl2 ). terminate the reaction by adding 3 mL of IKI reagent (0.6% iodine in 6% KI; 1 mL of this diluted to 50 mL with 0.05 N HCl), and record the OD at 620 nm. The amount of starch degraded (an estimate of enzyme activity) can be determined by a calibration curve.

Further, it can also be measured through agar plate assay in which the diffusion of a-amylase from seed tissue into an agar gel containing starch enabled assessment of enzyme activity through starch-IKI reaction.

Prepare 0.2% starch solution, add pinch of CaCl2 (0.2 mM CaCl2). Add agar (2%) in the above boiling solution, transfer the hot solution into petri plates and allow it to solidify. Thereafter, carefully place the transversely cut seed halves on the above agar plate. After incubation (vary from seed to seed), stain the agar plates with IKI reagent to determine amylase activity.

Sun, Z. and Henson, C.A., 1991. A quantitative assessment of the importance of barley seed α-amylase, β-amylase, debranching enzyme, and α-glucosidase in starch degradation. Archives of Biochemistry and Biophysics, 284(2), pp.298-305.