Enhancement of Potency of Streptokinase Isolated from Streptococcus pyogenes by Conjugation with Polyethylene glycol-200 Vaibhav Dagaji Aher1, R. Kavitha1, T. Sivakumar1, D. Krishnakumar1, Pronobesh Chattopadhay2, Subham Banerjee2, Arjun Patra3
1 Department of Pharmaceutical Biotechnology, Nandha College of Pharmacy, Tamilnadu, India
2 Cellular and Microbiology Laboratory, College of Pharmacy, Institute of Foreign Trade & Management, Uttar Pradesh, India
3 Department of Pharmacy, Banasthali University, Banasthali Vidyapith Rajasthan, India
Corresponding author: Vaibhav Dagaji Aher, Department of Pharmaceutical Biotechnology,
Nandha College of Pharmacy, Erode-638 052, Tamilnadu (India),
e-mail: vaibhav2020@gmail.com
Streptokinase is used as fibrinolytic agent to dissolve blood clots in deep vein thrombosis and is easily susceptible to proteolysis. The following study aimed to isolate streptokinase from Streptococcus pyogenes, to conjugate it with polyethylene glycol-200 (PEG 200) and to study the stability in presence of proteolytic enzymes. Streptococcus pyogenes (NCIM No. 2608) was cultivated in MRS (Man-Rogosa-Sharpe) broth. Streptokinase was isolated by gel filtration chromatography using Sephadex G-100 and further
purified by SDS-PAGE. The potency assay was performed by radial caseinolysis and Christenson two-fold serial dilution methods. Activity found was 252 IU/ml. Km and Vmax were 0.44 nm and 294 IU/ml, respectively. Isolated streptokinase was conjugated with PEG 200 which was confirmed by SDS-PAGE and FT-IR. Potency was increased to 310 IU/ml with Km and Vmax values of 0.59 nm and 337 IU/ml, respectively. Streptococcus pyogenes is an alternative source of streptokinase and conjugation with PEG 200 increases its potency.
Key words Polyethylene glycol • SDS-PAGE • Streptococcus pyogenes • Streptokinase |
