The organism isolated from an infected green leaf sample was selected based on the extracellular protease production using casein plate assay. The organism was identified as Cladosporium species. optimization of growth parameters using submerged fermentation, shake flask studies showed maximum enzyme activity at an inoculums size of 106 spores/ml, pH 9.0 (enzyme activity-228 U/ml/min), incubation temperature 28oC, incubation time 72 hours (enzyme activity-420 U/ml/min). The thrombolytic enzyme was partially purified up to 1.8 fold by 60% (NH4)2SO4 fractionation then subjected to simple dialysis followed by ion exchange chromatography. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) shows the partially purified thrombolytic enzyme at the molecular weight of ≈35kDa. The thrombolytic enzyme showed maximum activity at pH10 at 50oC. This work reveals the potential of Cladosporium species as an unconventional and unexplored production alternative to already known thrombolytic agents.
Hariharan P, Chandrashekhar Naik, Akanksha Viresh Sharma, Pooja Singh, Prerana Suresh Kumar, Azra Kausar, Revansiddappa. Isolation, production and purification of new thrombolytic enzyme from Cladosporium Spp.. European Journal of Biotechnology and Bioscience, Volume 6, Issue 2, 2018, Pages 01-04