Cloning, purification and characterization of a cellulase-free xylanase from Geobacillus thermodenitrificans AK53
- Authors: Irfan M.1, Guler H.I.2, Belduz A.O.3, Shah A.A.1, Canakci S.3
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Affiliations:
- Departmet of Microbiology, Faculty of Biological Sciences
- Department Departmet of Molecular Biology and Genetic, Faculty of Sciences
- Department of Biology, Faculty of Sciences
- Issue: Vol 52, No 3 (2016)
- Pages: 277-286
- Section: Article
- URL: https://journals.rcsi.science/0003-6838/article/view/151910
- DOI: https://doi.org/10.1134/S0003683816030066
- ID: 151910
Cite item
Abstract
Geobacillus thermodenitrificans AK53 xyl gene encoding xylanase was isolated, cloned and expressed in Escherichia coli. After purifying recombinant xylanase from G. thermodenitrificans AK53 (GthAK53Xyl) to homogeneity by ammonium sulfate precipitation and ion exchange chromatography, biochemical properties of the enzyme were determined. The kinetic studies for GthAK53Xyl showed KM value to be 4.34 mg/mL (for D-xylose) and Vmax value to be 2028.9 μmoles mg–1 min–1. The optimal temperature and pH for enzyme activity were found out to be 70°C and 5.0, respectively. The expressed protein showed the highest sequence similarity with the xylanases of G. thermodenitrificans JK1 (JN209933) and G. thermodenitrificans T-2 (EU599644). Metal cations Mg2+ and Mn2+ were found to be required for the enzyme activity, however, Co2+, Hg2+, Fe2+ and Cu2+ ions caused inhibitor effect on it. GthAK53Xyl had no cellulolytic activity and degraded xylan in an endo-fashion. The action of the enzyme on xylan from oat spelt produced xylobiose and xylopentose. The reported results are suggestive of a xylanase exhibiting desirable kinetics, stability parameters and metal resistance required for the efficient production of xylobiose at industrial scale.
About the authors
M. Irfan
Departmet of Microbiology, Faculty of Biological Sciences
Email: belduz@ktu.edu.tr
Pakistan, Islamabad
H. I. Guler
Department Departmet of Molecular Biology and Genetic, Faculty of Sciences
Email: belduz@ktu.edu.tr
Turkey, Trabzon, 61080
A. O. Belduz
Department of Biology, Faculty of Sciences
Author for correspondence.
Email: belduz@ktu.edu.tr
Turkey, Trabzon, 61080
A. A. Shah
Departmet of Microbiology, Faculty of Biological Sciences
Email: belduz@ktu.edu.tr
Pakistan, Islamabad
S. Canakci
Department of Biology, Faculty of Sciences
Email: belduz@ktu.edu.tr
Turkey, Trabzon, 61080
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