Development of recombinant Bacillus subtilis producing β-glucanase enzymes as a fish feed additive

In this thesis, the recombinant pRS416G vector carrying the β-(1,3)-glucanase gene from Escherichia coli bacterium was transformed into Bacillus subtilis BR151 by electrotransformation technique. It was observed that as a result of electrotransformation, they formed recombinant colonies containing laminarin plates. On this plate, recombinant colonies showed positive zones with Congo-red. The protein band of 125 kDa responsible for the β-(1,3)-glucanase enzyme is shown in SDS-PAGE and zymogram analyzes. On the other hand, recombinant vector pNW33N/β-(1,3-1,4)-glucanase gene, carrying the β-(1,3-1,4)-glucanase gene, was transformed into Bacillus subtilis RSKK245 strain by electrotransformation technique. Enzymatic activity regions recombinant B. subtilis RSKK245 colonies were shown by straining with Congo-red. The protein band of the enzyme with a size of 26 kDa was shown by zymogram analysis. These results show that the cloned genes are expressed into B. subtilis strain successfully.