Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli
In thisstudy Bacillus subtilis akky1 strainwas isolated from the soil of beech forest in Akkuş City, Ordu Province,Turkey. The identification of the strain akky1 done by PCR amplification 16SrRNA. The full-length 16S rRNA sequence showed the highest nucleotidesimilarity (100%) with Bacillus subtilis...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Sakarya University
2018-12-01
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| Series: | Sakarya Üniversitesi Fen Bilimleri Enstitüsü Dergisi |
| Subjects: | |
| Online Access: | https://dergipark.org.tr/tr/download/article-file/381057 |
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| Summary: | In thisstudy Bacillus subtilis akky1 strainwas isolated from the soil of beech forest in Akkuş City, Ordu Province,Turkey. The identification of the strain akky1 done by PCR amplification 16SrRNA. The full-length 16S rRNA sequence showed the highest nucleotidesimilarity (100%) with Bacillus subtilisstrain B7 (KC310823.1). Spesific oligonucleotides targeting the sequence of Bacillus subtilis xylanase gene given inGenBank were used to amplify a 642-bp fragment from genomic DNA. The geneencoding xylanase was cloned into pET28b (+) plasmid vector, sequenced andexpressed in Escherichia coli BL21(DE3). The hexahistidine (6xHis) tagged fusion protein was purified usingnickel affinity chromatography and the xylanase activity was measured. Themolecular mass of the purified xylanase was approximately 26 kDa as estimatedby SDS-PAGE. The xylanase had optimal activity at pH 6.0 and 60°C.The Km values of the recombinant enzyme towards beechwood was 3,33mg/ml. |
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| ISSN: | 2147-835X |