Experimental Preliminary Study for Production of Recombinant Subtilisin Enzyme by pET28b Cloning Vector

Abstract

Enzymes are biological catalysts and allow reactions to occur 103 – 108 times faster than reactions without enyzmes. Because of these, they are widely used in industry and the use of proteases accounts for 60 % of all enyzmes. It is quite hard and costly to obtain from biological resources. Therefore, it is more attractive to produce them by recombinant DNA technology that enables the production of many proteins. Subtilisin is a serine protease enzyme and it has a broad range of applications in detergent, food, textile, pharmaceutical industries. Thus, its continuous production has a large importance. In this study originated from this point; subtilisin gene belonging to Bacillus subtilis was cloned in pET28b vector and then expression was performed. Subtilisin enzyme was produced and it was clearly observed in the 35000-55000 Da band range by SDS-PAGE analysis, which has 41646.82 Da molecular mass.

Keywords

• Recombinant Protein
• Subtilisin
• pET28b

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