ASSESSMENT OF ALPHA-AMYLASE INHIBITOR IN SEEDS OF COMMON PULSES AND ITS PARTIAL PURIFICATION
Five Pulses were purchased from local market and analysis for protein content, carbohydrate and total phenolic content was done. All the Pulses were treated with Amylase Inhibitor to check its activity. Highest activity was found in KID, and it was further purified for Dialysis. On purified KID-sample, SDS was performed to analyze and determine the amylase inhibitor activity.
The maximum protein content (192.880.03mg/g) was found in sample- KID and minimum protein content (1350.64mg/g) was found in sample-LEN. Carbohydrate content was determined in all pulses and maximum carbohydrate content (3460.07mg/g) was found in sample KID and minimum carbohydrate content (167.230.22) was found in sample-LEN. The maximum Total Phenolic Content was found in sample-KID (13.100.20mgGAE/g) and minimum was found in sample-PP (4.800.56mgGAE/g).After that the Amylase inhibitory activity was calculated and maximum activity was seen in sample-KID (70.020.84%) and minimum in sample-PP (39.430.47%). The effect of Temperature on alpha-amylase inhibitor showed that the inhibitory activity of sample-KID was found to be 70.450.15% and it increases up to 60C (78.230.65) and after that it decreases. Upon partial purification of Alpha-Amylase Inhibitor, the crude extract showed the inhibitory activity of 70.67% and then specific activity further increased in Dialyzed and salt precipitated sample and that was 85.25%.
Upon SDS weight of partially purified sample was compared to standard protein marker (14.2-96.4kd). The molecular weight of partially purified sample was found to be in between the range of 17-20kd, suggesting that amylase inhibitor is of low molecular weight.