Screening of Micromycetes with Ability to Produce Proteases with Human Activated Protein C-Like Activity

Activated protein C (APC) is a main protease player of a human anticoagulant system. Acting on factors of haemostatic system, APC prevents intravascular blood coagulation. Low level of APC increases risk of thrombosis that’s why it is extremely important to develop new methods of treatment and diagnosis of it’s deficiency. For this purpose in medical practice proteases with protein C activating and APC-like activities are used. However, often such enzymes are expensive and insufficiently specific; therefore a search of new such enzymes is so much actual.

It is demonstrated that micromycetes are perspective source of proteases with blood-like activities. A protease of  Aspergillus ochraceus with protein C activating  activity wasinvestigated and characterized, nevertheless it isstill no data about enzymes with APC-like activity.

Sixteen cultures of micromycetes were checked up on their ability to produce enzymes with APC-like activity and then physiology and dynamic of enzyme production of selected culture was studied.

Our research demonstrated that purification of enzymes with direct APC-like action is not widespread among micromycetes. Significant APC-like activity was registered only for one strain of Purpureocillum lilacinum. An activity of culture liquidwith chromogenic peptide substrate of APC on a third day at 28oC was25.84 Е/ml×10-3.

Following investigation of enzyme production dynamics of selected culture detected highest APC-like activity on 13 day of cultivation at 25оС. Value was 58.79 Е/ml×10-3.

Importantly that action of detected enzyme is probably specific as we can see by no reaction with a number of different chromogenic peptide substrates. Low collagenolytic and fibrinolytic activities also positively distinguish this protein as a potential medicine. Thereby a further study of this protein is required.