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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Sakarya University
2018-12-01
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| Series: | Sakarya Üniversitesi Fen Bilimleri Enstitüsü Dergisi |
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| Online Access: | https://dergipark.org.tr/tr/download/article-file/381057 |
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| author | Sema Bilgin Yakup Ulusu Hülya Kuduğ İsa Gökçe |
| author_facet | Sema Bilgin Yakup Ulusu Hülya Kuduğ İsa Gökçe |
| author_sort | Sema Bilgin |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-62316591b4a9445db9870fcc47c3d6d0 |
| institution | Kabale University |
| issn | 2147-835X |
| language | English |
| publishDate | 2018-12-01 |
| publisher | Sakarya University |
| record_format | Article |
| series | Sakarya Üniversitesi Fen Bilimleri Enstitüsü Dergisi |
| spelling | doaj-art-62316591b4a9445db9870fcc47c3d6d02024-12-23T07:59:09ZengSakarya UniversitySakarya Üniversitesi Fen Bilimleri Enstitüsü Dergisi2147-835X2018-12-012261508151710.16984/saufenbilder.32715328Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coliSema Bilgin0Yakup Ulusu1Hülya Kuduğ2İsa Gökçe3GAZİOSMANPAŞA ÜNİVERSİTESİ, FEN-EDEBİYAT FAKÜLTESİ, KİMYA BÖLÜMÜKARAMANOĞLU MEHMETBEY ÜNİVERSİTESİ, MÜHENDİSLİK FAKÜLTESİ, BİYOMÜHENDİSLİK BÖLÜMÜGAZİOSMANPAŞA ÜNİVERSİTESİ, MÜHENDİSLİK VE DOĞA BİLİMLERİ FAKÜLTESİGAZİOSMANPAŞA ÜNİVERSİTESİ, MÜHENDİSLİK VE DOĞA BİLİMLERİ FAKÜLTESİIn 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.https://dergipark.org.tr/tr/download/article-file/381057bacillus subtilisksilanazrekombinant proteinendüstriyel enzimlerescherichia colibacillus subtilisxylanaserecombinant proteinindustrial enzymesescherichia coli |
| spellingShingle | Sema Bilgin Yakup Ulusu Hülya Kuduğ İsa Gökçe Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli Sakarya Üniversitesi Fen Bilimleri Enstitüsü Dergisi bacillus subtilis ksilanaz rekombinant protein endüstriyel enzimler escherichia coli bacillus subtilis xylanase recombinant protein industrial enzymes escherichia coli |
| title | Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli |
| title_full | Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli |
| title_fullStr | Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli |
| title_full_unstemmed | Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli |
| title_short | Cloning, Expression and Characterization of Xylanase (xyn-akky1) from Bacillus subtilis in Escherichia coli |
| title_sort | cloning expression and characterization of xylanase xyn akky1 from bacillus subtilis in escherichia coli |
| topic | bacillus subtilis ksilanaz rekombinant protein endüstriyel enzimler escherichia coli bacillus subtilis xylanase recombinant protein industrial enzymes escherichia coli |
| url | https://dergipark.org.tr/tr/download/article-file/381057 |
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