Abstract
The alkaline xylanase Xyn11A-LC from the alkalophilic Bacillus sp. SN5 was expressed in E. coli, purified and crystallized. The crystal structure was determined at a resolution of 1.49 Å. Xyn11A-LC has the β-jelly roll structure typical of family 11 xylanases. To improve its thermostability and thermophilicity, a mutant SB3 was constructed by introducing three arginines on the different sides of the protein surface. SB3 increased the optimum temperature by 5 °C. The wild type and SB3 had the half-lives of 22 and 68 min at 65 °C at pH 8.0 (Tris/HCl buffer), respectively. CD spectroscopy revealed that the melting temperature (T m) of the wild type and SB3 were 55.3 and 66.9 °C, respectively. These results showed that the introduction of arginines enhance the thermophilicity and thermostability of Xyn11A-LC.
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Acknowledgments
This study was supported by the National Basic Research Program of China (2011CBA00800 and 2009CB724700), Chinese National Programs for High Technology Research and Development (2011AA02A206 and 2012AA022100) and the Knowledge Innovative Program of Chinese Academy of Science (KSCX2-EW-G-8).
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Supplementary Table 1: Data collection and refinement statistics for Xyn11A-LC
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Bai, W., Zhou, C., Xue, Y. et al. Three-dimensional structure of an alkaline xylanase Xyn11A-LC from alkalophilic Bacillus sp. SN5 and improvement of its thermal performance by introducing arginines substitutions. Biotechnol Lett 36, 1495–1501 (2014). https://doi.org/10.1007/s10529-014-1512-7
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DOI: https://doi.org/10.1007/s10529-014-1512-7