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Journal of Polymers and the Environment

, Volume 27, Issue 12, pp 2895–2905 | Cite as

An Eco-friendly Method of Purification for Xylanase from Aspergillus niger by Polyelectrolyte Precipitation

  • María Victoria Podestá
  • Esteban Amador Morilla
  • María Belén Allasia
  • Nadia Woitovich Valetti
  • Gisela Tubio
  • María Julia BoggioneEmail author
Original paper
  • 24 Downloads

Abstract

The separation of xylanase (Xyl) from a fungal extract obtained by submerged fermentation (SmF) was carried out using precipitation with natural polyelectrolytes ɩ-carrageenan (Carr) and chitosan (CHS). Xyl activity determined from the fungal extract at different times was modeled by mixed linear model for longitudinal data considering the factors as fixed effects and the experimental units as random effects. A spline regression was used to model the Xyl activity with knots at days 2 and 3 and interaction between three factors (p < 0.0001 in each section). High purification factors were reached with both polymers. Mass spectrometry analysis and zymogram analysis revealed that the Xyl that precipitated with both polymers had a molecular weight of 24 kDa and an isoelectric point (pI) of 5.45. The purification factor (PF) of Xyl in the precipitate of the fungal extract from SmF with 0.05% w/v CHS (pH 8.00) was around 5.6 for a mass ratio of 1.8 × 10−5 mg CHS/mg protein and with 0.5% w/v Carr (pH 7.00) it was 9.0 for a mass ratio of 0.03 mg Carr/mg protein. These results make this precipitation method suitable to be included in a downstream process, providing such advantages as simplicity, scalability, ability to concentrate the enzyme in a single step process as well as the use non-toxic reagents. Taking into account that the process of purification of macromolecules is a decisive step with a profound impact on their final market cost, the developed methodology has a high potential to be used in the purification of Xyl.

Keywords

Xylanase Polyelectrolyte Carrageenan Chitosan 

Notes

Acknowledgements

This work was supported by the Grants from Agencia Nacional de Promoción Científica y Tecnológica (PICT 2016–1170 and 1712/17), from CONICET (PIP 505/15) and from the Secretary of Science and Technology of National University of Rosario (1BIO589 and BIO520). Authors would like to thank to Antonela Taddia for her colaboration and the Staff from the English Department (Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario), for correcting the language of this manuscript.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.IPROByQ-CONICET, Faculty of Biochemical and Pharmaceutical SciencesNational University of RosarioRosarioArgentina
  2. 2.Area of Statistics and Data Processing, Faculty of Biochemical and Pharmaceutical SciencesNational University of RosarioRosarioArgentina

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