OPTIMIZATION OF PHYSICAL PARAMETERS AND CHEMICAL PRE-TREATMENT IMPROVES THE SOLUBILITY OF PROTEASE A, A RECOMBINANT PROTEOLYTIC ENZYME
DOI:
https://doi.org/10.53808/KUS.2010.10.1and2.1013-LKeywords:
Protein solubility, physical parameters, ultrafiltration concentrate, enzyme concentrationAbstract
Cost of total process of enzyme production at industrial scale depends mainly on its recovery and the recovery largely depends on the solubility of enzymes in solution. Protease A, a proteolytic enzyme produced by genetically engineered Bacillus licheniformis is included in the detergents used for dish washing. In the present study, we have studied the influence of physical parameters and chemical pretreatment on the solubility of Protease A in detail. In case of temperature, a longer protein solubility region as a function of pH and dry matter percentage was found at 4°C for Protease A than that of ambient temperature. The maximum dry matter percentage was found 20.4% RI at pH 4.0. The optimum protein solubility for Protease A was found with in a conductivity range, 4.0 to 5.0 mS/cm within a pH values 4.0 to 6.0 with the highest clarity. The solubility region of Protease A with respect to enzyme concentration was found within the range of 81.0 to 35.4 mg/gm over a large range of pH starting from 4.0 to 8.0. The solubility trend of the protein was found not to be changed by the level of the enzyme concentration. Findings of this study followed the theory related to protein solubility. Pre-treatment with Proxel, an antimicrobial agent increased the solubility of Protease A as a function of pH. This study clearly demonstrates that optimization of some physical parameters and chemical pre-treatment improves the solubility of Protease A and thus establishes the conditions of parameters in a cost-effective strategy of Protease A recovery and production at industrial scale.
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