| Abstract: | Aminopeptidase is isolated and purified from the culture liquid of the thermophilic strain of Bacillus licheniformis. The aminopeptidase predominantly splits off N-terminal leucin in short peptides and hydrolyzes leucinamide as well. The molecular weight of the enzyme is about 60 kDa. The enzyme is able to form aggregates. Optimum of aminopeptidase activity was demonstrated at pH 8.0-8.3 and temperature of 85 degrees C. The enzyme is inactivated by metal-binding reagents and reducing substances, and is activated by cobalt and PCMB ions. The EDTA-inactivated enzyme activity is reduced by cobalt and zinc ions, however the latter has no activating action. The enzyme under study is characterized by high thermostability: in the presence of the substrate at the temperature of 90 degrees C the reaction linearity is retained for not less than 2 h and without the substrate the half-life of the aminopeptidase at 90 degrees C is 145 min. Extracellular aminopeptidase of the thermophilic strain of B. licheniformis is a new enzyme differing from the aminopeptidases described by the present in high thermostability, induced, evidently, by the presence of one or several disulphide bonds in the enzyme molecule. |
