For Biotechnological Applications: Purification and Characterization of Recombinant and Nanoconjugated Xylanase Enzyme from Thermophilic Bacillus Subtilis

Abstract

This study aimed to produce and nanoconjugated the xylanase enzyme of Bacillus subtilis BTX6 (MH101286) isolated from geothermal hot springs in Diyadin, Agri (Davud). Within the scope of this study, the 1,4-beta-endo xylanase (GH11 family) free enzyme from B. subtilis was cloned, recom-binantly expressed in E. coli after that nanoconjugated by encapsulation with the Anadoluca method. Then, the recombinantly produced enzyme and the nanoconjugated enzyme were char-acterized. According to the results, the optimum activity of both recombinant enzyme and nanoconjugated xylanase enzyme were determined as pH 7.0. Considering the optimal tempera-ture values, it was determined that the recombinant enzyme display optimum activity at 68 degrees C, while the nanoconjugated enzyme shows the best activity at 75 degrees C. The molecular weight of the recombinantly produced enzyme was measured as 71 kDa. In the enzyme activity measurements, the recombinant enzyme was determined as 1803 U/mg., and the activity of the nanoconjugated enzyme was determined as 1898 U/mg. Some metal ions such as MgSO4, CuSO4, CaCl2, ZnSO4, and FeSO4 increased the activity of recombinant and nanoconjugated enzymes. The Km value for the recombinant enzyme was 2.298 (mM), the Vmax value was 5.691 (U/mg.). Nanoconjugated enzyme the enzyme increased Km (2.298-2.402 mM) and Vmax (5.691-6.195 U/mg.) values. As a result, it was concluded that the cloning and nanoconjugated xylanase enzyme methods preferred in this study can be used effectively to improve enzyme activity in industrial processes.