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Development of hemicellulolytic enzyme mixtures for plant biomass deconstruction on target biotechnological applications

  • Biotechnological products and process engineering
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Abstract

An essential step in the conversion of lignocellulosic biomass to ethanol and other biorefinery products is conversion of cell wall polysaccharides into fermentable sugars by enzymatic hydrolysis. The objective of the present study was to understand the mode of action of hemicellulolytic enzyme mixtures for pretreated sugarcane bagasse (PSB) deconstruction and wheat arabinoxylan (WA) hydrolysis on target biotechnological applications. In this study, five hemicellulolytic enzymes—two endo-1,4-xylanases (GH10 and GH11), two α-L-arabinofuranosidases (GH51 and GH54), and one β-xylosidase (GH43)—were submitted to combinatorial assays using the experimental design strategy, in order to analyze synergistic and antagonistic effects of enzyme interactions on biomass degradation. The xylooligosaccharides (XOSs) released from hydrolysis were analyzed by capillary electrophoresis and quantified by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC–PAD). Based on this analysis, it was possible to define which enzymatic combinations favor xylose (X1) or XOS production and thus enable the development of target biotechnological applications. Our results demonstrate that if the objective is X1 production from WA, the best enzymatic combination is GH11 + GH54 + GH43, and for xylobiose (X2) production from WA, it is best to combine GH11 + GH51. However, if the goal is to produce XOS, the five enzymes used in WA hydrolysis are important, but for PSB hydrolysis, only GH11 is sufficient. If the final objective is bioethanol production, GH11 is responsible for hydrolyzing 64.3 % of hemicellulose from PSB. This work provides a basis for further studies on enzymatic mechanisms for XOS production, and the development of more efficient and less expensive enzymatic mixtures, targeting commercially viable lignocellulosic ethanol production and other biorefinery products.

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Acknowledgments

We are grateful to FAPESP (The State of São Paulo Research Foundation) for its financial support (2012/18859-5). ARLD is a FAPESP fellow (2013/18910-3). This work was also financially supported by FAPESP 2008/58037-9 and by CNPq 475022/2011-4 and 310177/2011-1 (FMS). We would like to thank the entire team of the molecular biology laboratory (CTBE/CNPEM), in particular the technical support of Rodrigo F. Almeida, also Thabata M. Alvarez and Zaira B. Hoffmam for cloning the GH10 and GH51 enzymes, respectively.

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  1. Brazilian Bioethanol Science and Technology Laboratory (CTBE), Brazilian Centre of Research in Energy and Materials (CNPEM), Giuseppe Máximo Scolfaro Street 10000, Campinas, SP, 13083, Brazil

    Rosana Goldbeck, André R. L. Damásio, Thiago A. Gonçalves, Carla B. Machado, Douglas A. A. Paixão, Lúcia D. Wolf, Fernanda Mandelli, George J. M. Rocha, Roberto Ruller & Fabio M. Squina

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  1. Rosana Goldbeck
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  2. André R. L. Damásio
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  3. Thiago A. Gonçalves
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  5. Douglas A. A. Paixão
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  10. Fabio M. Squina
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Correspondence to Fabio M. Squina.

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Goldbeck, R., Damásio, A.R.L., Gonçalves, T.A. et al. Development of hemicellulolytic enzyme mixtures for plant biomass deconstruction on target biotechnological applications. Appl Microbiol Biotechnol 98, 8513–8525 (2014). https://doi.org/10.1007/s00253-014-5946-6

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  • DOI: https://doi.org/10.1007/s00253-014-5946-6

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