Construction, Expression, and Characterization of a Thermostable Xylanase |
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Authors: | Xiao-Yan Weng Jian-Yi Sun |
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Institution: | (1) Department of Biological Science, College of Life Science, Zhejiang University, Hangzhou, 310029, P.R., China;(2) Microbiology Division of Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, 310029, P.R., China |
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Abstract: | A hybrid gene, btx, encoding a thermostable xylanase, Btx, was constructed by substituting the 31 N-terminal amino acid residues of the Thermomonospora fusca xylanase A (TfxA) for the corresponding region of 22 amino acid residues of the Bacillus subtilis xylanase A (BsxA). The btx gene was expressed in Escherichia coli BL21. The halo size produced by xylanase Btx on a Remanzol brilliant blue R (RBB) xylan plate at 60°C and pH 6.0 was larger than those of BsxA and TfxA. The molecular weight of Btx was 22 kDa. Temperature and pH optima for Btx were at 50–60°C and 6.0, respectively. Btx showed activity over 80% over a pH range of 5.0–9.0, which was wider than that of BsxA, and was also more acid-resistant than TfxA. Btx exhibited significant thermostability compared with BsxA. The results show the importance of the N-terminal sequence of TfxA in thermostability. |
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