Influence of Enzyme Activators on Alkaline Protease from Thermo-Tolerant Bacillus licheniformis JX849145

Proteolytic enzymes particularly proteases, have become an important and essential part of the industrial processes including pharmaceuticals, food products, laundry and detergents. Metal ions are involved in many enzyme catalyzed reaction. Hence prior to this, supplementation of ten different metal ions improved substantially the growth of B. licheniformis JX849145 and protease yield. Among ten metal ions, the Ca2+ (3 mM), Mg2+ (2 mM) and Zn2+ (2 mM) exhibits maximum protease production. The presence of K+, Na+, Fe2+and Cu2+ ions in the medium resulted in significant reduction in both growth and production of protease. The integrated effect of metal ions was also investigated by Plackett-Burman design to find the mutual interaction of process variables. During growth of the cell, metal ions are required for stability of the cell at different physico chemical conditions. Therefore in present investigation the three optimized metal ions such as Mg2+, Ca2+ and Zn2+ were used to find the superior combination at variable concentration. Earlier findings showed that the Metal ions such as Zn2+ often have a structural role, while notably Ca2+ is necessary for regulation of the activity. The designed Plackett Burman runs exhibit, 1.20 U/mg protease activity at 2 mM concentration of metal ion (Mg2+, Ca2+ and Zn2+). The correlation in between 0.5, 1.0 and 2.0 mM concentration of metal ions was analyzed by regression equation and analysis of variance (ANOVA). The obtained statistics showed 0.000 of its p value, it indicates the used concentration of metal ions was significant. The result showed that the coefficient of determination (R2) was 98.5 % and R2 (adj) was 98.0 % which ensured satisfactory adjustment of the quadratic model to the experimental data. Alkaline protease obtained from B. licheniformis JX849145 has properties such as optimum temperature at 50C, optimal pH 10, stability towards metal ions and actively involved with commercial detergents that make this enzyme potentially useful for various industrial applications.