Introduction: Fibrinolytic enzymes must currently originate a significant product in the arena of medical research. Very limited studies are stated on fibrinolyticenzyme production from actinomycetes. Methods: Streptomyces sp. isolated from marine soil was chosen to optimize its fibrinolytic protease production.16s rRNA sequencing confirmed the isolated potent strain to be Streptomyces rubiginosus VITPSS1. A fibrinolytic protease was then purified from Streptomyces rubiginosus VITPSS1, with the target of producing a cost effective feasible enzyme from a potential actinomycete. Results: SDS-PAGE results exhibited a protein band of about 45 kDa and the fibrinolytic band was detected by zymography. Optimization of physical and nutritional parameters for fibrinolytic protease production from a marine soil isolate Streptomyces strain was done by response surface methodology. The optimal cultural condition for fibrinolytic protease production was obtained with response surface methodology was based on OFAT results, it was inferred that glycerol, Soyabean meal, pH 7.2 and temperature 37 ◦ C. The optimization of the production of fibrinolytic protease with response surface methodology bring about two-folds increase in production by Streptomyces rubiginosus VITPSS1. Conclusion: Thus, this study presents its novelty by highlighting the potential of marine Streptomyces as a significant source for fibrinolytic enzyme production. © 2018, Bentham Science Publishers B.V. All rights reserved.