Optimization of Activated Carbon Adsorption Detoxification of Sugarcane Bagasse Hydrolysate by Response Surface Methodology to Enhance Ethanol Production
DOI:
https://doi.org/10.69650/rast.2025.260766Keywords:
Detoxification, Activated Carbon, Sugarcane, Central Composite Design, Ethanol YieldAbstract
Lignocellulosic biomass has significant potential as a renewable resource for biofuel production. Bioethanol is a widely used biofuel derived from agricultural residues such as cassava, rice straw, sugarcane, and sorghum. In this study, sugarcane bagasse was used as the raw material for ethanol production by Pichia stipitis TISTR5806. The objective was to examine the effect of activated carbon in removing inhibitory compounds from hydrolysate to enhance ethanol fermentation. Sugarcane bagasse was pretreated with 2.0% v/v diluted sulfuric acid at 121°C for 60 minutes to obtain hydrolysate. Acid hydrolysis generates monomeric sugars and inhibitory compounds, including furfural, 5-hydroxymethylfurfural (5-HMF), phenolics, and organic acids. To mitigate the inhibitory effects and improve ethanol production, this study employed activated carbon for compound removal. A central composite design was used to determine the optimal detoxification conditions for activated carbon, varying its concentration (1–5% w/v), reaction temperature (30–60°C), and reaction time (20–60 minutes). The optimal conditions were identified as 3.0% w/v activated carbon, a reaction temperature of 45°C, and a reaction time of 40 minutes. Under these conditions, activated carbon removed 23.93% of total furans and 61.72% of phenolics. The ethanol yield from the untreated hydrolysate was 0.21 gproduct/gsubstrate, corresponding to a theoretical yield of 42.44%, with an ethanol production rate of 0.09 g/L·h. In contrast, the detoxified hydrolysate yielded 0.26 gproduct/gsubstrate, achieving a theoretical yield of 51.30% and an ethanol production rate of 0.28 g/L·h. These results indicate that sugarcane bagasse is a promising substrate for ethanol production and that activated carbon effectively removes inhibitory compounds from hydrolysate, thereby enhancing fermentation efficiency.
References
Sadigov, R., Rapid growth of the world population and its socioeconomic results. The Scientific World Journal. 2022 (2022) 8110229, doi: https://doi.org/10.1155/2022/8110229.
Ramaraj, R., Unpaprom, Y. and Dussadee, N. Cultivation of Green Microalga, Chlorella vulgaris for Biogas Purification. International Journal of New Technology and Research. 2 (2016) 117-122.
Hirani, A. H., Javed, N., Asif, M., Basu, S. K. and Kumar, A. in Biofuels: Greenhouse Gas Mitigation and Global Warming: Next Generation Biofuels and Role of Biotechnology: A Review on First- and Second-Generation Biofuel Productions, Ch. 8, Springer India, (2018) 141-154, doi: https://doi.org/10.1007/978-81-322-3763-1_8.
Chia, S. R., Chew, K. W., Show, P. L., Yap, Y. J., Ong, H. C., Ling, T. C. and Chang, J. S., Analysis of economic and environmental aspects of microalgae biorefinery for biofuels production: a review. Biotechnology journal. 13 (2018) 1700618, doi: https://doi.org/10.1002/biot.201700618.
Sjulander, N. and Kikas, T., Origin, impact and control of lignocellulosic inhibitors in bioethanol production—A review. Energies. 13 (2020) 4751, doi: https://doi.org/10.3390/en13184751.
Olsson, L. and Hahn-Hägerdal, B., Fermentation of lignocellulosic hydrolysates for ethanol production. Enzyme and Microbial technology. 18 (1996) 312-331, doi: https://doi.org/10.1016/0141-0229(95)00157-3.
Luo, X., Zeng, B., Zhong, Y. and Chen, J., Production and Detoxification of Inhibitors during the Destruction of Lignocellulose Spatial Structure. BioResources. 17 (2022) 1939-1961, doi: https://doi.org/10.15376/biores.17.1.Luo.
Saleh, S. H. and Abd Halim, N. in Fuel Ethanol Production from Sugarcane: Sugarcane Bagasse Pretreatment Methods for Ethanol Production. Ch. 4, IntechOpen, (2018) 63-80, doi: https://doi.org/10.5772/intechopen.81656.
Mosier, N., Wyman, C., Dale, B., Elander, R., Lee, Y. Y., Holtzapple, M. and Ladisch, M., Features of promising technologies for pretreatment of lignocellulosic biomass. Bioresource technology. 96 (2005) 673-686, doi: https://doi.org/10.1016/j.biortech.2004.06.025.
Jönsson, L. J., Alriksson, B. and Nilvebrant, N.-O., Bioconversion of lignocellulose: inhibitors and detoxification. Biotechnology for biofuels. 6 (2013) 1-10, doi: https://doi.org/10.1186/1754-6834-6-16.
Luo, X., Zeng, B., Zhong, Y. and Chen, J., Production and Detoxification of Inhibitors during the Destruction of Lignocellulose Spatial Structure. BioResources. 17 (2022) 1939-1961, doi: https://doi.org/10.15376/biores.17.1.Luo.
Singh, O. V., Silva, S. and Chandel, A. in Biofuel Production - Recent Developments and Prospects: Detoxification of Lignocellulosic Hydrolysates for Improved Bioethanol Production. Ch. 10, IntechOpen, (2011) 225-246, doi: https://doi.org/10.5772/16454.
Dąbrowski, A., Podkościelny, P., Hubicki, Z. and Barczak, M., Adsorption of phenolic compounds by activated carbon—a critical review. Chemosphere. 58 (2005) 1049-1070, doi: https://doi.org/10.1016/j.chemosphere.2004.09.067.
Haghseresht, F., Finnerty, J. J., Nouri, S. and Lu, G. Q., Adsorption of Aromatic Compounds onto Activated Carbons: Effects of the Orientation of the Adsorbates. Langmuir. 18 (2002) 6193-6200, doi: https://doi.org/10.1021/la025541b.
Huang, C.-F., Lin, T. -H., Guo, G. -L. and Hwang, W. -S., Enhanced ethanol production by fermentation of rice straw hydrolysate without detoxification using a newly adapted strain of Pichia stipitis. Bioresource technology. 100 (2009) 3914-3920, doi: https://doi.org/10.1016/j.biortech.2009.02.064.
Germec, M., Kartal, F. K., Bilgic, M., Ilgin, M., Ilhan, E., Güldali, H., Isci, A. and Turhan, I., Ethanol production from rice hull using Pichia stipitis and optimization of acid pretreatment and detoxification processes. Biotechnology progress. 32 (2016) 872-882, doi: https://doi.org/10.1002/btpr.2275.
Prasad, S., Malav, M. K., Kumar, S., Singh, A., Pant, D. and Radhakrishnan, S., Enhancement of bio-ethanol production potential of wheat straw by reducing furfural and 5-hydroxymethylfurfural (HMF). Bioresource Technology Reports. 4 (2018) 50-56, doi: https://doi.org/10.1016/j.biteb.2018.09.007.
Miller, G. L., Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical chemistry. 31 (1959) 426-428, doi: https://doi.org/10.1021/ac60147a030.
Nielsen, S. S. Phenol-sulfuric acid method for total carbohydrates. Nielsen, S.S. edn, Springer, Boston, MA, 2010, doi: https://doi.org/10.1007/978-1-4419-1463-7_6.
Martinez, A., Rodriguez, M. E., York, S. W., Preston, J. F. and Ingram, L. O., Use of UV absorbance to monitor furans in dilute acid hydrolysates of biomass. Biotechnology progress. 16 (2000) 637-641, doi: https://doi.org/10.1021/bp0000508.
Blainski, A., Lopes, G. C. and De Mello, J. C. P., Application and analysis of the folin ciocalteu method for the determination of the total phenolic content from Limonium brasiliense L. Molecules. 18 (2013) 6852-6865, doi: https://doi.org/10.3390/molecules18066852.
Myers, J. A., Curtis, B. S. and Curtis, W. R., Improving accuracy of cell and chromophore concentration measurements using optical density. BMC biophysics. 6 (2013) 1-15, doi: https://doi.org/10.1186/2046-1682-6-4.
Seo, H.-B., Kim, H.-J., Lee, O.-K., Ha, J.-H., Lee, H.-Y. and Jung, K.-H., Measurement of ethanol concentration using solvent extraction and dichromate oxidation and its application to bioethanol production process. Journal of industrial Microbiology and Biotechnology. 36 (2009) 285-292, doi: https://doi.org/10.1007/s10295-008-0497-4.
Martín, C., Galbe, M., Nilvebrant, NO. and Jönsson, L. J. Comparison of the fermentability of enzymatic hydrolyzates of sugarcane bagasse pretreated by steam explosion using different impregnating agents. Applied Biochemistry and Biotechnology. 98 (2002) 699-716, doi: https://doi.org/10.1385/abab:98-100:1-9:699.
Ravindran, R. and Jaiswal, A. K., A comprehensive review on pre-treatment strategy for lignocellulosic food industry waste: challenges and opportunities. Bioresource technology. 199 (2016) 92-102, doi: https://doi.org/10.1016/j.biortech.2015.07.106.
Hong, J.-W., Gam, D.-H., Kim, J.-H., Jeon, S.-J., Kim, H.-S. and Kim, J.-W., Process development for the detoxification of fermentation inhibitors from acid pretreated microalgae hydrolysate. Molecules. 26 (2021) 2435, doi: https://doi.org/10.3390/molecules26092435.
Tesfaw, A. A. and Tizazu, B. Z., Reducing sugar production from Teff straw biomass using dilute sulfuric acid hydrolysis: Characterization and optimization using response surface methodology. International Journal of Biomaterials. 2021 (2021) 2857764, doi: https://doi.org/10.1155/2021/2857764.
Yaya, O. L., Appiah, K. E. K., Doudjo, S., Nassirou, A. K. M., Didier, F. G., Benjamin, Y. K., Drogui, A. P. and Tyagi, D. R., Multi-response and multi-criteria optimization of acid hydrolyzate detoxification of cocoa pod husks: Effect on the content of phenolic compounds and fermentable sugars. Heliyon. 9 (2023) 1-11, doi: https://doi.org/10.1016/j.heliyon.2023.e15409.
Yildirim, O., Ozkaya, B., Altinbas, M. and Demir, A., Statistical optimization of dilute acid pretreatment of lignocellulosic biomass by response surface methodology to obtain fermentable sugars for bioethanol production. International Journal of Energy Research. 45 (2021) 8882-8899, doi: https://doi.org/10.1002/er.6423.
Gupta, V. C., Singh, M., Prasad, S. and Mishra, B. N., Minimization of Inhibitor Generation in Rice Straw Hydrolysate Using RSM Optimization Technique. Agriculture. 13 (2023) 1431, doi: https://doi.org/10.3390/agriculture13071431.
Deshavath, N. N., Mohan, M., Veeranki, V. D., Goud, V. V., Pinnamaneni, S. R. and Benarjee, T., Dilute acid pretreatment of sorghum biomass to maximize the hemicellulose hydrolysis with minimized levels of fermentative inhibitors for bioethanol production. 3 Biotech. 7 (2017) 1-12, doi: https://doi.org/10.1007/s13205-017-0752-3.
Baig, M. Z. and Dharmadhikari, S. M., Optimization of detoxification with over liming and charcoal treatment for increasing the fermentability of cotton stalk hydrolyzate. Indian Journal of Applied Research. 4 (2014) 453-455, doi: https://doi.org/10.15373/2249555x/july2014/143.
Srinorakutara, T., Subkaree, Y., Boonvitthya, N. and Bamrungchue, N., Optimum Condition of Rice Straw Hydrolysate Detoxification with Charcoal Powder for Cellulosic Ethanol Production by Pichia stipitis TISTR 5806. Journal of Food Science and Engineering. 6 (2016) 75-81.
Brito, P. L., de Azevedo Ferreira, C. M., Silva, A. F. F., Pantoja, L. d. A., Nelson, D. L. and dos Santos, A. S., Hydrolysis, detoxification and alcoholic fermentation of hemicellulose fraction from palm press fiber. Waste and Biomass Valorization. 9 (2018) 957-968, doi: https://doi.org/10.1007/s12649-017-9882-4.
Germec, M. and Turhan, I., Ethanol production from acid-pretreated and detoxified rice straw as sole renewable resource. Biomass Conversion and Biorefinery. 8 (2018) 607-619, doi: https://doi.org/10.1007/s13399-018-0310-1.
Taherzadeh, M. J. and Karimi, K. Biofuels: Alternative Feedstocks and Conversion Processes. 1st edn., Academic Press, 2011, doi: https://doi.org/10.1016/B978-0-12-385099-7.00012-7.
Palmqvist, E., Almeida, J. S. and Hahn‐Hägerdal, B., Influence of furfural on anaerobic glycolytic kinetics of Saccharomyces cerevisiae in batch culture. Biotechnology and bioengineering. 62 (1999) 447-454, doi: https://doi.org/10.1002/(sici)1097-0290(19990220)62:4%3C447::aid-bit7%3E3.0.co;2-0.
Chen, X., Zhai, R., Li, Y., Yuan, X., Liu, Z.-H. and Jin, M., Understanding the structural characteristics of water-soluble phenolic compounds from four pretreatments of corn stover and their inhibitory effects on enzymatic hydrolysis and fermentation. Biotechnology for biofuels. 13 (2020) 1-13, doi: https://doi.org/10.1186/s13068-020-01686-z.
Mussatto, S. I. and Roberto, I. C., Alternatives for detoxification of diluted-acid lignocellulosic hydrolyzates for use in fermentative processes: a review. Bioresource technology. 93 (2004) 1-10, doi: https://doi.org/10.1016/j.biortech.2003.10.005
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 School of Renewable Energy and Smart Grid Technology (SGtech)

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
All copyrights of the above manuscript, including rights to publish in any media, are transferred to the SGtech.
The authors retain the following rights;
1. All proprietary rights other than copyright.
2. Re-use of all or part of the above manuscript in their work.
3. Reproduction of the above manuscript for author’s personal use or for company/institution use provided that
(a) prior permission of SGtech is obtained,
(b) the source and SGtech copyright notice are indicated, and
(c) the copies are not offered for sale.




