In Vitro and Computational Evaluation of Thrombolytic Activity of Kinemakinase of Kinema, an Indigenous Fermented Food of Eastern Nepal

Kinema, an indigenous fermented soybean food originating from the eastern Himalayan regions of Nepal and India, contains a bacterium called Bacillus subtilis. This bacterium generates bioactive compounds and enzymes, some of which may have therapeutic uses. With cardiovascular diseases on the rise and the need for effective clot-dissolving agents, researchers decided to examine the clot-dissolving ability of a specific enzyme, kinemakinase, found in kinema made from white soybeans.

Through a process of extraction and partial purification using ammonium sulfate, the enzyme's clot-dissolving capabilities were tested in a laboratory setting using human blood clots. The results indicated that three dilutions of the enzyme successfully lysed the clots, accounting for 66% to 68% of the clot breakdown. To gain a deeper understanding of the enzyme's properties, in silico analyses were conducted, including modeling its tertiary structure using UniProt, refining it using GalaxyRefine, and docking it with fibrin to assess interaction and stability.

The use of disulfide engineering also proved beneficial for enhancing the protein's stability. The research concludes that kinema-derived kinemakinase could potentially serve as a valuable alternative thrombolytic agent, but further biochemical characterization and dosage optimization are required.