Applied Microbiology and Biotechnology

, Volume 87, Issue 6, pp 2125–2135 | Cite as

Functional analysis of glycoside hydrolase family 8 xylanases shows narrow but distinct substrate specificities and biotechnological potential

  • Annick Pollet
  • Jan Schoepe
  • Emmie Dornez
  • Sergei V. Strelkov
  • Jan A. Delcour
  • Christophe M. Courtin
Biotechnologically Relevant Enzymes and Proteins

Abstract

The potential of glycoside hydrolase family (GH) 8 xylanases in biotechnological applications is virtually unexplored. Therefore, the substrate preference and hydrolysis product profiles of two GH8 xylanases were evaluated to investigate their activities and substrate specificities. A GH8 xylanase from an uncultured bacterium (rXyn8) shows endo action but very selectively releases xylotriose from its substrates. It has a higher activity than the Pseudoalteromonas haloplanktis GH8 endo-xylanase (PhXyl) on xylononaose and smaller xylo-oligosaccharides. PhXyl preferably degrades xylan substrates with a high degree of polymerization. It is sterically more hindered by arabinose substituents than rXyn8, producing larger end hydrolysis products. The specificities of rXyn8 and PhXyl differ completely from these of the previously described GH8 xylanases from Bifidobacterium adolescentis (BaRexA) and Bacillus halodurans (BhRex). As reducing-end xylose-releasing exo-oligoxylanases, they selectively release xylose from the reducing end of small xylo-oligosaccharides. The findings of this study show that GH8 xylanases have a narrow substrate specificity, but also one that strongly varies between family members and is distinct from that of GH10 and GH11 xylanases. Structural comparison of rXyn8, PhXyl, BaRexA, and BhRex showed that subtle amino acid changes in the glycon as well as the aglycon subsites probably form the basis of the observed differences between GH8 xylanases. GH8 xylanases, therefore, are an interesting group of enzymes, with potential towards engineering and applications.

Keyword

Xylanase Glycoside hydrolase family 8 Substrate specificity Arabinoxylan Xylo-oligosaccharides 

Notes

Acknowledgements

The authors like to thank Dr. Charles C. Lee of the Agricultural Research Service (Albany, CA, USA) for donation of plasmid DNA from rXyn8 and Prof. Anna Kulminskaya from the Russian Academy of Science (St. Petersburg, Russia) for the labeled XOS. Financial support for the SBO IMPAXOS project by the ‘Instituut voor de aanmoediging van Innovatie door Wetenschap en Technologie in Vlaanderen’ (I.W.T., Brussels, Belgium), for the post-doctoral fellowship of E.D. by the ‘Fonds voor Wetenschappelijk Onderzoek-Vlaanderen’ (F.W.O, Brussels, Belgium) and from the Research Fund K.U.Leuven (GOA/03/10 and IDO/03/005) are gratefully acknowledged. S.V.S. thanks the Biomedical Sciences Group (K.U. Leuven) for a start-up grant. The study is part of the Methusalem program ‘Food for the Future’ at the K.U. Leuven.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Annick Pollet
    • 1
  • Jan Schoepe
    • 2
  • Emmie Dornez
    • 1
  • Sergei V. Strelkov
    • 2
  • Jan A. Delcour
    • 1
  • Christophe M. Courtin
    • 1
  1. 1.Laboratory of Food Chemistry and Biochemistry and Leuven Food Science and Nutrition Research Centre (LFoRCe)Katholieke Universiteit LeuvenLeuvenBelgium
  2. 2.Laboratory for BiocrystallographyKatholieke Universiteit LeuvenLeuvenBelgium

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