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Sequential pretreatment of sugarcane bagasse by alkali and organosolv for improved delignification and cellulose saccharification by chimera and cellobiohydrolase for bioethanol production

3 Biotech volume 11, Article number: 59 (2021) Cite this article

Abstract

Sequential pretreatments for sugarcane bagasse (scb) by NaOH followed by organosolv under mild conditions were evaluated for cellulose recovery and dilignification. The best-optimized sequential pretreatment of scb was obtained at 10% (w/v) of raw scb loading at 1% (w/v) NaOH (50 °C, 2 h) followed by treatment with organosolv (85%, v/v phosphoric acid, 50 °C, 1 h) with chilled acetone. This sequentially pretreated scb showed cellulose recovery, 66.1% (w/w) and delignification, 83.2% (w/w). NaOH or organosolv pretreated scb showed lower cellulose recovery 47.4% (w/w) or 54.5% (w/w) with lower delignification, 61% (w/w) or 56% (w/w), respectively. Pretreated solid residue of sequentially pretreated scb was enzymatically saccharified by chimera (β-glucosidase and endoglucanase, CtGH1-L1-CtGH5-F194A) and cellobiohydrolase (CtCBH5A) cloned from Clostridium thermocellum. Enzymatic hydrolysate of best sequentially pretreated scb gave total reducing sugar (TRS) yield, 230 mg/g and glucose yield, 137 mg/g pretreated scb. Only organosolv pretreated scb gave TRS yield, 112.5 mg/g and glucose yield, 72 mg/g of pretreated scb. Thus, sequentially pretreated scb resulted in 37% higher enzymatic digestibility than only orgnaosolv pretreated scb. Higher enzymatic digestibility was supported by higher crystallinity index CrI (45%) than those obtained with only organosolv pretreated (38%) or raw scb (25%). Field Emission Scanning Electron Microscope (FESEM) and Fourier-transform infrared (FT-IR) analyses showed enhanced cellulose exposure in sequentially pretreated scb. Preliminary investigation of bioethanol production at small scale by separate hydrolysis and fermentation (SHF) of enzymatic hydrolysate from best sequentially pretreated scb by Saccharomyces cerevisiae gave maximum ethanol yield of 0.42 g/g of glucose.

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Acknowledgements

Authors acknowledge the financial support provided by DBT-Pan-IIT Grant (BT/EB/PAN-IIT 2012), Centre for Bioenergy from Department of Biotechnology, Ministry of Science and Technology, New Delhi, India to Prof. Arun Goyal. The fellowship to Priyanka Nath was supported by funding from DST Inspire fellowship from Department of Science and Technology, New Delhi, India.

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Author notes

  1. Priyanka Nath and Premeshworii Devi Maibam have equally contributed to this work.

Affiliations

  1. Carbohydrate Enzyme Biotechnology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Priyanka Nath, Shweta Singh, Vikky Rajulapati & Arun Goyal

  2. Centre for Energy, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India

    Premeshworii Devi Maibam & Arun Goyal

  3. DBT PAN-IIT Center for Bioenergy, Indian Institute of Technology Guwahati, Guwahati, Assam, India

    Priyanka Nath, Shweta Singh & Arun Goyal

Authors
  1. Priyanka Nath
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  2. Premeshworii Devi Maibam
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  3. Shweta Singh
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  4. Vikky Rajulapati
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  5. Arun Goyal
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Contributions

AG conceived the idea and designed the objectives. PN, MPD, and SS performed the experiments. VR performed FT-IR analysis. AG, PN, MPD, and SS wrote the paper.

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Correspondence to Arun Goyal.

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Nath, P., Maibam, P.D., Singh, S. et al. Sequential pretreatment of sugarcane bagasse by alkali and organosolv for improved delignification and cellulose saccharification by chimera and cellobiohydrolase for bioethanol production. 3 Biotech 11, 59 (2021). https://doi.org/10.1007/s13205-020-02600-y

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  • DOI: https://doi.org/10.1007/s13205-020-02600-y

Keywords

  • Lignocellulosic biomass
  • Enzyme cocktail
  • Enzymatic hydrolysis
  • Bioethanol