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Study on Isolation, Optimization and Immobilization of Alkaline Protease produced by Aspergillus Flavus

Roshni Choubey*,Ram Keerti Verma, Ravi Prakash Mishra and Pooja Sharma

This study was conducted to isolate alkaline protease producing fungus, its identification and optimization of cultural conditions for production of alkaline protease enzyme. Preliminary screening was done on gelatin agar medium. Secondly gelatin liquification test was carried out to check the production of proteases functional at alkaline pH. The study related to process development involves optimization of different fermentation conditions towards enhancement of enzyme production for which the cultural conditions (physical and nutritional factors) during solid state fermentation for alkaline protease production by the isolated Aspergillus flavus were undertaken. Different types of substrates were selected for growing Aspergillus flavus. Alkaline protease production was found to be highest at temperature (280C), pH 8, incubation time 168 hrs, with metal ions (Mn), nitrogen source (peptone), and carbon source (sucrose) as the biggest zone of hydrolysis. The highest optical density and protein concentration was obtained at above mentioned conditions. The partial purification of alkaline protease enzyme from the culture filtrate was performed by ammonium sulfate salt precipitation which was immobilized using sodium alginate immobilization technique. Washing test resulted in complete removal of blood stain which indicates this enzyme can be proved crucial for detergent industry.