Amylase inhibitors of pigeonpea ( Cajanus cajan) seeds

Pigeonpea seeds were found to contain four human salivary amylase inhibitors (AIs), of which major AI have pI value about 6.2, AIs were heat labile, synthesized and degraded late during seed development and germination, and found to be moderately toxic against Helicoverpa armigera larvae in combination with protease inhibitors Pigeonpea ( Cajanus cajan L) seeds were analysed quantitatively for amylase inhibitor (AI) activity and qualitatively, by an in-gel-detection method on polyacrylamide gels. At least four AI isoforms were identified in pigeonpea seeds. The AIs inhibit human salivary and bovine pancreatic amylase but fail to inhibit bacterial, fungal and endogenous amylase. Pigeonpea AIs were found to be active over a pH range of 4.5 to 9.5 and were heat labile. The isoelectric point of a major inhibitor is 6.2. AIs were tolerant to proteolysis by trypsin, chymotrypsin, bromelain and endogenous pigeonpea proteases. Pigeonpea AIs were synthesized during late seed development and also degraded during late germination. Addition of AIs or protease inhibitors (PIs) alone to a diet of Helicoverpa armigera larvae did not increase mortality. However, the larvae reared on a diet containing AIs and PIs in combination, showed increased mortality and adverse effects on larval growth and development. In vitro inhibition of Helicoverpa gut amylase revealed that only 22% activity is sensitive to inhibitors. Further investigations on interactions of pigeoppea AIs and PIs with Helicoverpa gut enzymes is necessary to develop strategies to strengthen defense mechanisms in pigeonpea against H. armigera