Construction and application of recombinant strain of Bacillus subtilis for production of alkaline-protease

Protease enzymes are biocatalyst that has long been used as alternative to chemical reactions to improve the efficiency and cost-effectiveness of a wide range of industrial systems and bioprocesses. They are currently applied in both basic and applied arenas of research as well as in a wide range of product design and manufacturing processes e.g. those pertaining to the food, detergent, beverage, leather processing, waste water treatment, and in pharmaceutical industries. Until today, the largest share of the enzyme market has been held by a specific type of protease called alkaline proteases which are active and stable over a wide alkaline pH range. Since, alkaline protease production from naturally occurring wild type bacterial strains cannot meet the growing market demand due to its low yield and incompatibility with the standard industrial fermentation media formulations as well as with the fermentative conditions required for optimal growth of the host microorganisms. Remarkably, the enzyme production could be enhanced by taking the advantages of modern molecular biology techniques or recombinant DNA technology, whereby, the use of homologous or heterologous overexpression of genes coding for the target enzyme can be achieved for higher productivity in shorter time and that would reduce the costs of production as well. Therefore, in this work we will adopt a molecular approach to develop a new recombinant Bacillus strain, its improvement and growth optimization for the alkaline protease production using sustainable agri-industrial residues as feedstock carbon source. We will also investigate on the growth and process optimization of the culture during fermentation and other physiochemical parameters up to proto scale production level and its downstream purification to recover the final product. Hence, this study will provide a new perspective for the future bioprocess technology development and commercial success of an industrially important enzyme.