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Heterologous hyper-expression of a glucansucrase-type glycosyltransferase gene

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Abstract

Heterologous expression of the large glucansucrase-type glycosyltransferases genes is still a challenge, and typically yields are poor. Therefore, a number of different Escherichia coli systems for the expression of such a gene, encoding the glycosyltransferase R (GtfR) from Streptococcus oralis, were constructed and evaluated. We thereby obtained a strain producing the highest molar yields described so far for this class of enzymes. Cloning of a 5′-terminally truncated version of the gene in the expression vector pET33b(+) yielded, in dissolved form, about 2 μmol (300 mg) of enzyme per liter of culture of an optical density at 600 nm of four. Problems frequently encountered in the heterologous biosynthesis of this class of enzymes, such as formation of a high fraction of insoluble aggregates and/or proteolytic degradation, were not observed in the described system. The over-produced enzyme, devoid of almost its entire variable region, retained its characteristic activities.

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Acknowledgments

The authors thank Prof. K. Buchholz, Technical University of Braunschweig, for introducing us to this family of enzymes and for helpful discussions. Special thanks go to Dr. T. Fujiwara for his kind gift of plasmid pTH275. Financial support of this project by the Deutsche Forschungsgemeinschaft through SFB 578 (“From gene to product”), project A2, is gratefully acknowledged.

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Author notes

  1. Wera Collisi & Bernd Hofer

    Present address: Department of Chemical Biology, Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124, Braunschweig, Germany

  2. Anna Maria Swistowska

    Present address: Buck Institute for Age Research, Novato, CA, USA

Authors and Affiliations

  1. Division of Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany

    Anna Maria Swistowska, Sabine Wittrock, Wera Collisi & Bernd Hofer

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  1. Anna Maria Swistowska
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  2. Sabine Wittrock
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  3. Wera Collisi
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  4. Bernd Hofer
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Correspondence to Bernd Hofer.

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Swistowska, A.M., Wittrock, S., Collisi, W. et al. Heterologous hyper-expression of a glucansucrase-type glycosyltransferase gene. Appl Microbiol Biotechnol 79, 255–261 (2008). https://doi.org/10.1007/s00253-008-1435-0

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  • DOI: https://doi.org/10.1007/s00253-008-1435-0

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