Abstract
The gene, xynB, from Streptomyces olivaceoviridis A1 encoding xylanase, XYNB, with a high specific activity for xylan, was transformed into potato (Solanum tuberosum L.) by Agrobacterium tumefaciens. The integration of xynB into genomic DNA was confirmed by PCR and reverse transcriptase–PCR. The gene was expressed under the control of a constitutive double cauliflower mosaic virus (CaMV) 35S promoter. Both SDS-PAGE and western blot analysis showed high levels of expression of the 21 kDa and 31 kDa XYNB proteins in transgenic potato plants transformed by the binary vectors pBinXy and signal peptide contained pBinSPXy, respectively. The recombinant XYNB protein was present at up to 5% of total soluble leaf protein in the cytoplasm. In transgenic leaf and tuber extracts, xylanase activity was up to 87 μmol min−1 g−1 fresh leaf (9.7 μmol min−1 mg−1 total soluble protein). The xylanase was stable at 60°C and 70°C in buffers (pH 5.2) for 5 min. Furthermore, the xylanase enzymatic activity remained virtually unchanged over several generations of potato. These results demonstrate that the transgenic potato can be used to produce recombinant xylanase with high specific enzyme activity and can potentially be an alternative to present-day xylanase additives to animal feed.
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Acknowledgements
This work was funded by the Chinese National High Technology Research and Development Program (863 Program, Grant No. 2003AA214030) and the Chinese National Transgenic Research and Development Program (Grant No. JY03-A-16-02).
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Yang, P., Wang, Y., Bai, Y. et al. Expression of xylanase with high specific activity from Streptomyces olivaceoviridis A1 in transgenic potato plants (Solanum tuberosum L.). Biotechnol Lett 29, 659–667 (2007). https://doi.org/10.1007/s10529-006-9280-7
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DOI: https://doi.org/10.1007/s10529-006-9280-7