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Bu calismada, Bacillus subtilis akky1 straini Turkiye’nin Ordu Ili’nin Akkus ilcesindeki kayin ormanindan alinan topraktan izole edilmistir. akky1 straini 16S rRNA analizi ile tanimlanmistir. 16S rRNA dizisi Bacillus subtilis strain B7 (KC310823.1) ile %100’luk bir benzerlik gostermistir. GenBank’da verilen Bacillus subtilis ksilanazina ait gen dizisi esas alinarak tasarlanan primerler kullanilarak genomic DNA’dan 642 bp’lik bir DNA fragmenti elde edilmistir. Ksilanazi kodlayan gen, pET28b (+) ekspresyon vektorune klonlanmis ve Escherichia coli BL21 (DE3)’de ekpresse edilmistir. Rekombinant olarak uretilen protein nikel afinite kromatografisi ile saflastirilmis ve ksilanaz aktivitesi belirlenmistir. Saflastirilan ksilanazin molekuler agirligi SDS-PAGE ile 26.0 kDa olarak belirlenmistir. Rekombinant enzimin optimum pH’si 6.0 ve sicakligi 60°C, Km degeri ise 3.33 mg/ml olarak tespit edilmistir.

Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli