Purification, physicochemical properties, and statistical optimization of fibrinolytic enzymes especially from fermented foods: A comprehensive review

Fibrinolytic enzymes are proteases responsible for cleavage of fibrin mesh in thrombus clots, which are the primary causative agents in cardiovascular diseases. Developing safe, effective and cheap thrombolytic agents are important for prevention and cure of thrombosis. Although a wide variety of sources have been discovered for fibrinolytic enzymes, only few of them have been employed in clinical and therapeutic applications due to the drawbacks such as high cost of production, low stability of enzyme or therapeutic side effects. However, the discovery of new fibrinolytic enzymes requires complex purification stages and characterization, which gives an insight into their diverse modes of action. Post-discovery, approaches such as a) statistical optimization for fermentative bioprocessing and b) genetic engineering are advantageous in providing economic viability by finding simple and cost-effective medium, strain development with sufficient nutrient supplements for stable and high-level production of recombinant enzyme. This review provides a comprehensive understanding of different sources, purification techniques, production through genetic engineering approaches and statistical optimization of fermentation parameters as proteases have a wide variety of industrial and biotechnological applications making 60% of total enzyme market worldwide. New strategies targeting increased enzyme yields, non-denaturing environments, improved stability, enzyme activity and strain improvement have been discussed.