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Effect of enzymatic pretreatment of sugarcane bagasse with recombinant hemicellulases and esterase prior to the application of the cellobiohydrolase CBH I Megazyme®

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Abstract

The polysaccharides in the primary plant cell wall are a renewable energy source for biofuel production. However, these polysaccharides are not readily available for bioconversion, and large enzyme sets are required to deconstruct them. Here, we aimed to improve the glucan conversion using recombinant hemicellulases and esterase as a treatment in exploded and sugarcane bagasses (SCB), followed by the addition of commercial CBH I to prevent its inhibition by hemicellulases products. A high secretion level of the recombinant enzymes was observed on SDS-PAGE. The highest activities were verified at a temperature and pH ranging from 40 to 55 °C and 4.5 to 6.0, respectively. The released reducing sugar analysis showed that all enzymes act better on SCB, with xylanase C (XynC) presenting the best activity (0.54 U/mg of protein). The high-performance liquid chromatography (HPLC) analysis demonstrated that 24 h of pretreatment was enough to reach maximum glucan conversion. The best synergy was achieved between XynC and CBH I on SCB, 1.4%. All results showed that the enzymes acted better on SCB, which can be related to the biomass composition and its molecular structure. The enzymatic pretreatment of SCB with XynC was essential to improve the glucan conversion by CBH I.

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Acknowledgments

We are thankful for Dr. George Jackson de Moraes Rocha, from the Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), who gave us the biomass used in this research. Dr. Mark R. Wilkins and Dr. Michael Mueller cordially provided laboratory support and help with the HPLC analyses that were performed in the Biosystems and Agricultural Engineering department at the Oklahoma State University.

Funding

This work was funded by the Brazilian CNPq fellowship (Grant No. 141133/2009-0) to RCL and the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) post-doctoral fellowship to TBO (Grant No. 2017/09000-4).

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Authors and Affiliations

  1. Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    Rosymar Coutinho de Lucas, Thiago Machado Pasin, Ana Sílvia de Almeida Scarcella & Maria de Lourdes Teixeira de Moraes Polizeli

  2. Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    Rosymar Coutinho de Lucas, Tássio Brito de Oliveira & Maria de Lourdes Teixeira de Moraes Polizeli

  3. Department of Biology, University of Western Ontario, London, ON, Canada

    Matheus Sanitá Lima

  4. Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA

    Rolf Alexander Prade

  5. Departamento de Biotecnologia da Escola de Engenharia de Lorena, Universidade de São Paulo, Lorena, SP, Brazil

    Fernando Segato

Authors
  1. Rosymar Coutinho de Lucas
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  2. Tássio Brito de Oliveira
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  3. Matheus Sanitá Lima
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  4. Thiago Machado Pasin
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  6. Rolf Alexander Prade
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  7. Fernando Segato
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  8. Maria de Lourdes Teixeira de Moraes Polizeli
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Correspondence to Rosymar Coutinho de Lucas or Maria de Lourdes Teixeira de Moraes Polizeli.

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de Lucas, R.C., de Oliveira, T.B., Lima, M.S. et al. Effect of enzymatic pretreatment of sugarcane bagasse with recombinant hemicellulases and esterase prior to the application of the cellobiohydrolase CBH I Megazyme®. Biomass Conv. Bioref. 12, 491–499 (2022). https://doi.org/10.1007/s13399-020-00719-9

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