Fibrinolytic enzymes are agents that dissolve fibrin clots. These fibrinolytic agents have potential use to treat cardiovascular diseases, such as heart attack and stroke. In the present article, a fibrinolytic enzyme producing Pseudoalteromonas sp. IND11 was isolated from the fish scales and optimized for enzyme production. Cow dung was used as a substrate for the production of fibrinolytic enzyme in solid-state culture. A two-level full factorial design was used for the screening of key ingredients while further optimization was carried out using the central composite design. Statistical analysis revealed that the second-order model is significant with model F-value of 6.88 and R2 value of 0.860. Enzyme production was found to be high at pH 7.0, and the supplementation of 1% (w/w) maltose and 0.1% (w/w) sodium dihydrogen phosphate enhanced fibrinolytic enzyme production. The optimization of process parameters using response surface methodology resulted in a three-fold increase in the yield of fibrinolytic enzyme. This is the first report on production of fibrinolytic enzyme using cow dung substrate in solid-state fermentation.