Abstract
A trypsin inhibitor from seeds of faba bean (Vicia faba L.) was purified to near homogeneity as judged by native-PAGE with about 11 % recovery using ammonium sulphate fractionation, ion-exchange chromatography on DEAE-cellulose and gel filtration through Sephadex G-100. The inhibitor had a molecular weight of 18 kD as determined by SDS-PAGE and Sephadex G-100. The inhibitor inhibited trypsin and chymotrypsin to the extent of 48 and 12 %, respectively. The inhibtion was of non-competitive type with dissociation constant for the enzyme inhibitor complex in the region of 0.07 mg·ml−1. The inhibtor was stable between pH 4 and 5. It completely lost its activity when heated at 125 °C for 1 h or at 100 °C for 2 h. The inhibitor also lost its activity on exposure to 2-mercaptoethanol. Based on these properties, it could be concluded that Vicia faba trypsin inhibitor belongs to Bowman-Birk type of inhibitors, as it has molecular weight lower than generally observed for Kunitz type inhibitors.
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Abbreviations
- PAGE:
-
polyacrylamide gel electrophoresis
- DEAE:
-
diethyl amino ethyl
- SDS:
-
sodium dodecyl sulphate
- kD:
-
kilodalton
- CM:
-
carboxy methyl
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Gupta, P., Dhawan, K., Malhotra, S.P. et al. Purification and characterization of trypsin inhibitor from seeds of faba bean (Vicia faba L.). Acta Physiol Plant 22, 433–438 (2000). https://doi.org/10.1007/s11738-000-0085-3
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DOI: https://doi.org/10.1007/s11738-000-0085-3