@article{article_327153, title={Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli}, journal={Sakarya University Journal of Science}, volume={22}, pages={1508–1517}, year={2018}, DOI={10.16984/saufenbilder.327153}, author={Bilgin, Sema and Ulusu, Yakup and Kuduğ, Hülya and Gökçe, İsa}, keywords={Bacillus subtilis,Xylanase,Recombinant Protein,Industrial Enzymes,Escherichia coli}, abstract={<p class="MsoNormal" style="margin-right:14.35pt;"> <span lang="en-gb" xml:lang="en-gb">In this
study <i> Bacillus subtilis </i> akky1 strain
was isolated from the soil of beech forest in Akkuş City, Ordu Province,
Turkey. The identification of the strain akky1 done by PCR amplification 16S
rRNA. The full-length 16S rRNA sequence showed the highest nucleotide
similarity (100%) with <i>Bacillus subtilis </i>
strain B7 (KC310823.1). Spesific oligonucleotides targeting the sequence of <i>Bacillus subtilis </i>xylanase gene given in
GenBank were used to amplify a 642-bp fragment from genomic DNA. The gene
encoding xylanase was cloned into pET28b (+) plasmid vector, sequenced and
expressed in <i>Escherichia coli </i>BL21
(DE3). The hexahistidine (6xHis) tagged fusion protein was purified using
nickel affinity chromatography and the xylanase activity was measured. The
molecular mass of the purified xylanase was approximately 26 kDa as estimated
by SDS-PAGE. The xylanase had optimal activity at pH 6.0 and 60 <sup>° </sup>C.
The K <sub>m </sub> values of the recombinant enzyme towards beechwood was 3,33
mg/ml. </span> </p> <p> </p>}, number={6}, publisher={Sakarya University}