The α-glucuronidase Agu1 from Schizophyllum commune is a member of a novel glycoside hydrolase family (GH115)

  • Sun-Li ChongEmail author
  • Evy Battaglia
  • Pedro M. Coutinho
  • Bernard Henrissat
  • Maija Tenkanen
  • Ronald P. de VriesEmail author
Biotechnologically Relevant Enzymes and Proteins


Schizophyllum commune produces an α-glucuronidase that is active on polymeric xylan, while the ascomycete α-glucuronidases are only active on xylan oligomers. In this study, we have identified the gene (agu1) encoding this enzyme and confirmed the functionality by overexpression of the gene in S. commune and degradation of aldopentauronic acids, (MeGlcA)3-Xyl4, in the cultivation medium of the transformants. Expression analysis demonstrated that agu1 is not co-regulated with the predominant xylanase-encoding gene (xynA) of S. commune. The detailed sequence analysis of Agu1 demonstrated that this gene belongs to a novel glycoside hydrolase family (GH115) that also contains candidate genes from ascomycete fungi and bacteria. Phylogenetic analysis showed that the fungal GH115 α-glucuronidases are distinctly separate from the prokaryotic clade and distributed over three branches. The identification of putative genes encoding this enzyme in industrial fungi, such as Aspergillus oryzae and Hypocrea jecorina, will provide a starting point for further analysis of the importance of this enzyme for the hydrolysis of plant biomass.


α-Glucuronidase Schizophyllum commune GH115 Gene expression Xylan degradation 



We thank Prof. Dr. Han Wösten for access to the S. commune genome sequence, Prof. Annele Hatakka (Department of Food and Environmental Sciences, University of Helsinki) for providing the S. commune growing facilities, Dr. Kristiina Mäkinen (Department of Food and Environmental Sciences, University of Helsinki) for providing the agu1 cDNA isolation facilities, Dr. Matti Siika-aho (VTT Technical Research Centre) and Dr. Sanna Koutaniemi (Department of Food and Environmental Sciences, University of Helsinki) for the purification of S. commune α-glucuronidase (for raising antibody for Western blotting), Dr. Luis G. Lugones (Microbiology, University of Utrecht) for useful suggestions for the growing of S. commune, and Dr. Karin Scholtmeijer (Microbiology, University of Utrecht) for the expression vector. The financial support from the Academy of Finland through the WoodWisdom-Net Programme (HemiPop project no. 1124281), Glycoscience Graduate School (S.-L. C.), and COST 928 and COST FP0602 Short Term Scientific Mission (STSM) are gratefully acknowledged. E. B. and R. P. V. were supported by the Dutch Technology Foundation STW, the applied science division of NWO, and the Technology Program of the Ministry of Economic Affairs (project no. 07063).

Supplementary material

253_2011_3157_MOESM1_ESM.doc (36 kb)
Supplemental Fig. 1 Nucleotide and translated amino acid sequences of S. commune agu1. Exons are shown in upper case letters and introns are shown in lower case letters. The signal peptide is highlighted and the N-glycosylation sites are underlined accordingly (DOC 36 kb)
253_2011_3157_MOESM2_ESM.doc (32 kb)
Supplemental Fig. 2 Alignment of Agu1, the N-terminal amino acid sequence of the purified α-glucuronidase from S. commune (Tenkanen and Siika-aho 2000), and the second GH115 member identified in the S. commune genome. Identical amino acids between the N-terminal sequence of the purified enzyme and Agu1 are underlined. Identical amino acids between Agu1 and the second GH115 member are indicated by stars. This alignment demonstrates that Agu1 encodes the purified enzyme and that the two GH115 members differ significantly from each other throughout the amino acid sequence (DOC 32 kb)
253_2011_3157_MOESM3_ESM.doc (198 kb)
Supplemental Fig. 3 Bootstrap-supported phylogenetic analysis of family GH115, showing all of the fungal proteins formed a group and separated from bacterial proteins. The characterized enzymes from P. stipitis (Scheffersomyces stipitis CBS 6054; Ryabova et al. 2009) and S. commune (this study) are indicated in bold type. Bootstrap values for the respective branches are shown (DOC 198 kb)
253_2011_3157_MOESM4_ESM.xls (28 kb)
Supplemental Table 1 Genes used in the alignment (XLS 28 kb)


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Sun-Li Chong
    • 1
    Email author
  • Evy Battaglia
    • 2
  • Pedro M. Coutinho
    • 3
  • Bernard Henrissat
    • 3
  • Maija Tenkanen
    • 1
  • Ronald P. de Vries
    • 2
    • 4
    Email author
  1. 1.Department of Food and Environmental Sciences, Faculty of Agriculture and ForestryUniversity of HelsinkiHelsinkiFinland
  2. 2.Microbiology and Kluyver Centre for Industrial FermentationUtrecht UniversityUtrechtThe Netherlands
  3. 3.AFMB—UMR 6098, CNRSUniversités Aix-Marseille I and II, Case 932Marseille Cedex 9France
  4. 4.CBS-KNAW Fungal Biodiversity CentreUtrechtThe Netherlands

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