Poultry Feed Enrichment Using Covalently Immobilized Partially Purified Alkaline Protease from Newly Isolated Ochrobactrum anthropi KTP9

Proteases represent 60% of the enzyme market and around 66% of the proteases are derived from the microbial sources. In the present study, protease enzyme was produced from bacterial isolate Ochrobactrum anthropi KTP9 under submerged fermentation having 17.43 IU/mL activity and 3.626 mg/ml of protein content. Ammonium sulphate precipitation increased the specific activity from 4.80 to 22.45 IU/ ml with 4.67 purification fold. Free protease has optimum pH 8.0, temperature 35 °C and found stable upto 28 days on storage at 4 °C. The immobilization yield and immobilization efficiency were found as 40.54% and 89.72%, respectively. Upon immobilization, shift in temperature (35 to 45 °C) and pH optima (8.0 to 10.0) was observed as compared to free protease. The immobilized protease was found to retain 50% residual activity upto 35 days of storage at 4 °C. Immobilization enzyme can be reused upto 6 cycles with 50% residual activity. The immobilized protease showed 76% protein hydrolysis in 120 min as compare to free KTP9 protease.