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Optimization of Dilute Acid Pretreatment for Enhanced Release of Fermentable Sugars from Sugarcane Bagasse and Validation by Biophysical Characterization

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Abstract

Pretreatment of biomass is one of the most challenging steps in the process of second-generation (2G) ethanol and biochemical production. Dilute acid pretreatment is a widely adapted and convenient method to recover pentose (C5) as well as hexose (C6) sugars due to its featured solubilization of hemicellulose and cellulose before and after enzymatic saccharification, respectively. In the present study, dilute sulfuric acid (H2SO4) pretreatment of sugarcane bagasse (SCB) was statistically optimized using the face-centered composite design (FCCD) of response surface methodology (RSM) in terms of acid concentration (0.1–3% v/v), solid loading (5–20% w/v) and residence time (15–60 min) at constant temperature of 121 °C followed by enzymatic hydrolysis using commercial cellulase (Novozymes Cellic CTec2) for enhanced combined sugar yield (CSY) comprising of C5 and C6 sugars in pretreated as well as saccharified hydrolysates. Optimized process parameters found in the study were 2.18% (v/v) acid; 14.35% (w/v) solid loading; and 29.49 min residence time. CSY under optimized conditions was found to be 521.42 ± 7.2 g/kg raw SCB with 72.06 ± 1.0% sugars recovered out of the maximum theoretical sugars present in the raw biomass. Total reducing sugar yields in pretreated and saccharified hydrolysates were found to be 215.28 ± 2.4 and 306.14 ± 5.3 g/kg raw SCB, respectively. Morphological and structural changes in optimized pretreated and saccharified biomass further validated the efficiency of optimized pretreatment applied in the present study. The maximum ethanol concentration, volumetric productivity and yield from released sugars were calculated as 10.82 ± 2.2 g/L, 0.45 ± 0.9 g/L/h and 0.42 g/g-glucose consumed or 71.45 ± 2.5 g/kg raw SCB, respectively. Ethanol yield obtained from the fermentation of dilute H2SO4-pretreated SCB was corresponding to 82.4% of the theoretical ethanol yield.

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Acknowledgements

Authors (Meenu Hans and Sachin Kumar) are very thankful to the Department of Biotechnology, Ministry of Science and Technology, India, for providing funds to carry out the research work via grant no. DBT/IC-2/Indo-Brazil/2016-19/05. One of the authors (Meenu Hans) is very thankful to Sardar Swaran Singh National Institute of Bio-Energy, Kapurthala for providing Senior Research Fellowship, and Guru Nanak Dev University, Amritsar for providing Ph.D. registration (Reg. No. 2007.KJ/A.519). We also acknowledge Novozymes, Denmark, for providing Cellic Ctec 2. Biophysical study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) via grants 405191/2015-4, 140667/2015-6, 158752/2015-5, 303988/2016-9 and 440977/2016-9 and by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) via grant#2015/13684-0. A.K. Chandel is thankful to the CAPES for visiting researcher and professor program. The SEM experiments were carried out with the assistance of Mr. Manoel Ricardo Roncon, São Carlos Institute of Physics, University of São Paulo.

Funding

Department of Biotechnology, Ministry of Science and Technology, India (DBT/IC-2/Indo-Brazil/2016–19/05); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (405191/2015–4, 140667/2015–6, 158752/2015–5, 303988/2016–9 and 440977/2016–9); and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (2015/13684–0).

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Authors and Affiliations

  1. Biochemical Conversion Division, Sardar Swaran Singh National Institute of Bio-Energy, Kapurthala, India, 144601

    Meenu Hans & Sachin Kumar

  2. São Carlos Institute of Physics, University of São Paulo, Av. Trabalhador São-carlense, 400, Parque Arnold Schimidt, São Carlos, SP, 13566-590, Brazil

    Meenu Hans, Vanessa O. A. Pellegrini, Jefferson G. Filgueiras, Eduardo R. de Azevedo, Francisco E. C. Guimaraes & Igor Polikarpov

  3. Department of Microbiology, Guru Nanak Dev University, Amritsar, Punjab, 143005, India

    Meenu Hans & Bhupinder Singh Chadha

  4. Department of Biotechnology, Engineering School of Lorena (EEL), University of São Paulo, Lorena-SP, Brazil

    Anuj Kumar Chandel

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  1. Meenu Hans
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Contributions

MH conceived, planned, executed experiments and wrote original draft; VOAP, JGF, ERA and FECG analyzed biophysical parameters; AKC edited; IP reviewed and supervised; BSC reviewed; and SK conceived, planned, reviewed, edited and supervised.

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Correspondence to Sachin Kumar.

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Hans, M., Pellegrini, V.O.A., Filgueiras, J.G. et al. Optimization of Dilute Acid Pretreatment for Enhanced Release of Fermentable Sugars from Sugarcane Bagasse and Validation by Biophysical Characterization. Bioenerg. Res. 16, 416–434 (2023). https://doi.org/10.1007/s12155-022-10474-6

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