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Construction and functional characterization of truncated versions of recombinant keratanase II from Bacillus circulans

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Abstract

There is a need for degradative enzymes in the study of glycosaminoglycans. Many of these enzymes are currently available either in their natural or recombinant forms. Unfortunately, progress in structure-activity studies of keratan sulfate (KS) have been impeded by the lack of a commercially available endo-β-N-acetylglucosaminidase, keratantase II. The current study uses a recently published sequence of a highly thermostable keratanase II identified in Bacillus circulans to clone and express a series of truncation mutants in Escherichia coli BL21. The resulting truncated forms of keratanase II exhibit activity and excellent storage and thermal stability making these useful tools for glycobiology research.

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Acknowledgements

This research was funded in part by a grant from the National Institutes of Health HL094463 (RJL), HL125371 (RJL, FZ), HL136271 (RJL) and by the Chinese Study Fellowship (HW).

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Author notes

  1. Haisheng Wang and Wenqin He contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Chinese Academy of Agricultural Sciences, People’s Republic of China, Beijing, 100081, China

    Haisheng Wang

  2. Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Haisheng Wang, Wenqin He, Mattheos Koffas, Fuming Zhang & Robert J. Linhardt

  3. Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Peixia Jiang, Yanlei Yu, Lei Lin, Xiaojun Sun & Robert J. Linhardt

  4. Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, 100101, China

    Peixia Jiang

  5. Departments of Biology, Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Robert J. Linhardt

Authors
  1. Haisheng Wang
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  2. Wenqin He
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  3. Peixia Jiang
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  4. Yanlei Yu
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  5. Lei Lin
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  6. Xiaojun Sun
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  7. Mattheos Koffas
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  8. Fuming Zhang
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  9. Robert J. Linhardt
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Corresponding authors

Correspondence to Fuming Zhang or Robert J. Linhardt.

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The authors declare that they have no conflicts of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Wang, H., He, W., Jiang, P. et al. Construction and functional characterization of truncated versions of recombinant keratanase II from Bacillus circulans . Glycoconj J 34, 643–649 (2017). https://doi.org/10.1007/s10719-017-9786-3

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  • DOI: https://doi.org/10.1007/s10719-017-9786-3

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