Biotechnology Letters

, Volume 37, Issue 12, pp 2419–2426 | Cite as

Xanthomonas campestris expansin-like X domain is a structurally disordered beta-sheet macromolecule capable of synergistically enhancing enzymatic efficiency of cellulose hydrolysis

  • Atílio Tomazini Junior
  • Luciano Graciani Dolce
  • Mario de Oliveira Neto
  • Igor Polikarpov
Original Research Paper
First Online:
Received:
Accepted:

Abstract

Objectives

To biochemically characterize an expansin-like X protein domain from Xanthomonas campestris (XcEXLX1) and to study its synergy with cellulases in cellulose depolymerization.

Results

The protein was purified using a combination of ion exchange and size exclusion chromatography rendering about 30 mg pure protein/l culture medium. Circular dichroism spectroscopy and small-angle X-ray scattering studies of XcEXLX1 reveal that it is a strongly disordered β-sheet protein. Its low resolution envelope fits nicely the crystallographic structure of the homologous protein EXLX1 from Bacillus subtillis. Furthermore, we demonstrate that XcEXLX1 shows a synergistic, pH-dependent effect when combined with a commercial enzymatic preparation (Accellerase 1500), enhancing its hydrolytic activity on a cellulosic substrate. The strongest effect was observed in acid pHs with an increase in sugar release of up to 36 %.

Conclusion

The synergistic effect arising from the action of the expansin-like protein was considerable in the presence of significantly larger amounts of the commercial enzymatic cocktail then previously observed (0.35 FPU of Accellerase 1500/g substrate).

Keywords

Accellerase Biofuel Cellulase Cellulose depolymerization Expansin synergism Xanthomonas campestris 
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Notes

Acknowledgments

We thank Livia Regina Manzine, Andressa Alves Pinto, Derminda I. de Moraes, Joci Neuby Alves Macedo, Ethel Schuster and José Luis Lopes for the technical support. This research was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) via research Grants # 2008/56255-9, 2007/08706-9, 2010/52362-5 and 2009/05349-6, by Coordenacão de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) via INCT do Bioetanol and Grant # 471834/2009-2, 301981/2011-6 and 550931/2011-2.

Supporting information

Supplementary methods. Construction of the expression plasmid, Expression of the protein and its purification process.

Supplementary Fig. 1—Aligned sequences of gene (1) Xcc3535, (2) 2BH0 (BsEXLX1) and (3) 2HCZ (expansin of Zea mays), using the multiple alignment Clustal program (Higgins and Sharp 1988).

Supplementary Fig. 2—Size-exclusion chromatogram of XcEXLX1.

Supplementary material

10529_2015_1927_MOESM1_ESM.doc (176 kb)
Supplementary material 1 (DOC 176 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Atílio Tomazini Junior
    • 1
  • Luciano Graciani Dolce
    • 1
  • Mario de Oliveira Neto
    • 2
  • Igor Polikarpov
    • 1
  1. 1.Departamento de Física e Ciência Interdisciplinar, Instituto de Física de São CarlosUniversidade de São PauloSão CarlosBrazil
  2. 2.Instituto de BiociênciasUniversidade Estadual PaulistaBotucatuBrazil

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