# Enhanced fibrinolytic enzyme production by Oidiodendron maius through green bioprocessing of agro-industrial residue

## TL;DR

This study shows that mutagenizing Oidiodendron maius with ethidium bromide improves fibrinolytic enzyme production, which could help treat blood clots more effectively.

## Contribution

The study introduces a novel method of enhancing fibrinolytic enzyme production in Oidiodendron maius using ethidium bromide mutagenesis and agro-industrial residue.

## Key findings

- Ethidium bromide-treated O. maius mutants showed 1642.24 U/mg specific activity, much higher than the wild-type strain.
- Optimal enzyme activity was achieved at 35°C and pH 7.5 after parameter optimization.
- Wheat bran was identified as an effective substrate for enhanced enzyme production.

## Abstract

Thrombosis denotes the formation of blood clots within arteries and veins, representing a primary etiological factor in cardiovascular diseases often culminating in fatal outcomes. Prompt resolution of thrombotic disorders is achieved through expedited fibrinolysis facilitated by the administration of fibrinolytic enzymes, which constitute the optimal therapeutic approach. This research aimed to enhance the production of fibrinolytic enzymes through the cultivation of indigenously isolated strains of Oidiodendron maius using physical and chemical mutagenesis techniques. Fibrinolytic enzyme activity from mutant strains was validated by enzyme assays followed by purification using ammonium sulfate precipitation, desalting, ion exchange chromatography, gel filtration chromatography, and SDS-PAGE. Various kinetic and thermodynamic parameters were systematically optimized to maximize enzyme activity. In O. maius, the ethidium bromide mutant strain showed better results as compared to the other mutants with specific activity of 1642.24 U/mg and 0.5 mg/mL protein content compared to the wild-type strain which 90.20 U/mg specific activity and 3.7 mg/mL protein content. The optimum temperature and pH were 35°C and 7.5, respectively. The findings indicated that treating O. maius with ethidium bromide resulted in the generation of better mutants with enhanced enzyme activities compared to wild-type and other mutant strains. With optimization of multiple parameters, these strains demonstrate significant potential for enhanced fibrinolytic enzyme production by the usage of wheat bran as substrate.