Аннотация:We characterized an alkaline chymotrypsin-like serine peptidase from the yellow mealworm Te-nebrio molitor with a non-canonical substrate-binding subsite for its possible application as a component (anadditive) in various biological products. The enzyme was obtained as a recombinant preparation. Purificationwas carried out using affinity chromatography on Ni2+-NTA agarose. The specificity constants (kcat/KM) forthe chymotrypsin substrates, Glp-AAF-pNA, Suc-AAPF-pNA, and Ac-Y-pNA, were 7, 4.2, and0.9 (μM min) –1 , respectively. The optimum of the proteolytic activity was observed at pH 9.0. The enzymewas stable in the alkaline pH range, and in the presence of BSA, also in the acidic region. Peptidase was inhi-bited by synthetic inhibitors such as PMSF, TPCK, and chymostatin, while EDTA, E-64, and pepstatin hadno effect on the enzyme activity. The purified enzyme showed high stability over time in the presence of BSA.The short life cycle of the insect and the production of a large number of peptidases in the midgut with highcatalytic activity and stability can make T. molitor an excellent alternative source of industrially importantenzymes for application as components (additives) in various biological products (e.g., stain removers, deter-gents, etc.).
