Abstract: The present study has been conducted to isolate the potential fungal isolates in proteolysis process. One isolate showed high activity in protease production, being Mucor sp. Morphological and microscopical studies, as well as 18S rRNA sequence confirmed that this isolate is identified as Mucor mucedo strain SEE1 and sequencing product was deposited in the GenBank database under the accession number KP736529. Optimization of protease production during Plackett-Burman and central composite designs has been carried out, with six nutritional variables. Glucose and casein were significantly superior to other variables in protease production. Moreover, an overall 3.36-fold increase in protease production has been obtained by Placket-Burman. However, 1.14-fold increase was found in central composite design. On the other hand, during the fermentation process in optimized medium, liberated amino acids has been determined (µg/ml), where histidine came in the first order (1155.3) followed by lysine (897.4), tyrosine (634), arginine (580) and isoleucine (580), respectively. However, cysteine was found in less appreciable amount (36).