Crinumin, a chymotrypsin-like but glycosylated serine protease from Crinum asiaticum: Purification and physicochemical characterisation

Plant latex could be a potential source of novel proteases usable in the food and feed industries because of broad substrate specificity with high stability in extreme conditions. Crinumin, a glycosylated serine protease with chymotrypsin-like activity was purified from the latex of Crinumasiaticum using cation-exchange column chromatography. Crinumin shows activity over a wide range of pH (4.5–11.5 and optimum at 8.5), temperature (75 °C and optimum at 70 °C) and is also functional against chaotrophs, organic solvents, and detergents, even after prolonged exposure. The molecular mass (67.7 kDa), extinction coefficient (17.7), isoelectric point (6.9), and numbers of tryptophan (13), tyrosine (24) and cysteine (15 with 7 disulphide bridges) residues were estimated. Km of the enzyme was 31.7 μM with casein and 5 × 104 μM with N-succinyl-l-phenylalanine-p-nitroanilide. Easy availability of the aqueous latex, simple purification procedure, high yield (33%), stability and activity in adverse conditions makes it applicable for the pharmaceutical and food industries.