Construction of feed additive fish probiotics with genetic engineering techniques

In this thesis, β-Glucan which is a good source of feed additive fish probiotics, was used in order to benefit from feed quality in fishery products, to increase live weight gain and to strengthen the immune system. β-Glukan is an important component of the cell walls of grains such as oats and barley. 1,3-1,4-β-D-glucan 4-glucanhydrolase (β-glucanase, lichenase) (EC 3.2.1.73) is an enzyme with the ability to hydrolyze β-glucans. In this study, recombinant vector pNW33N carrying the β (1,3-1,4) glucanase (lichenase) gene of Streptococcus bovis genome was transferred to Bacillus subtilis RSKK246 (CMCase +) strain by electroporation. Subsequently, electrotransformation waas performed on LB-agar plates containing lichenan and enzymatic activity regions of recombinant B. subtilis RSKK246 colonies were observed by staining with congo red. In addition, the DNA from the recombinant vector pNW33N/Lichenase was cut on both the BamHI and HindIII endonucleases and observed on the ligase gene (1800 bp) agarose gel. On the other hand, the total extracellular proteins of the recombinant protein band corresponding to the recombinant enzyme 26 kDa was observed by zymogram analysis. These results indicate that the β (1,3-1,4) glucanase gene has been successfully expressed to the B. subtilis strain RSKK246.