Low-Power Ultrasound-Assisted Methanesulfonic Acid–Formic Acid Pretreatment Coupled with Simultaneous Saccharification and Fermentation for Bioethanol Production from Sugarcane Leaves
Abstract
This study investigates an energy-efficient and environmentally sustainable approach to enhancing bioethanol production from sugarcane leaves through synergistic pretreatment using low-power ultrasound and an organic acid co-solvent system. To reduce the energy demand and environmental impact associated with conventional pretreatment methods, methanesulfonic acid (MSA) and formic acid (FA) were combined under low-power ultrasonic conditions. A Box–Behnken design (BBD) based on response surface methodology (RSM) was employed to optimize the FA concentration, ultrasonication time, and ultrasound power. The optimal conditions were identified as 29.17% FA, 18.70 min of ultrasonication, and an ultrasound power of 16.61 W, with the MSA concentration maintained at 5%. These conditions promoted the disruption of the lignocellulosic structure through acoustic cavitation and acid-assisted modification, thereby enhancing cellulose accessibility while minimizing energy consumption. Subsequent simultaneous saccharification and fermentation (SSF) using Saccharomyces cerevisiae at 32 °C with an enzyme loading of 50 FPU/g achieved an ethanol yield of 73.22% of the theoretical yield (on
a cellulose basis), corresponding to an ethanol concentration of 12.602 mg/mL after 48 h. Extending the fermentation time to 72 h resulted in only a marginal increase in ethanol yield of 0.8%, suggesting that a 48 h incubation period represents a potentially techno-economically optimal operating window by reducing processing time and associated production costs. Compared with separate hydrolysis and fermentation (SHF), SSF achieved a comparable ethanol concentration while shortening the overall processing time and eliminating the need for a separate hydrolysis step at 50 °C. These findings demonstrate that low-power ultrasound-assisted MSA–FA co-solvent pretreatment
is a promising strategy for converting sugarcane leaf residues into bioethanol under mild and potentially
cost-effective operating conditions.
Keywords
[1] S. Pan, H. M. Zabed, Y. Wei, and X. Qi, “Technoeconomic and environmental perspectives of biofuel production from sugarcane bagasse: Current status, challenges and future outlook,” Industrial Crops and Products, vol. 188, Nov. 2022, Art. no. 115684, doi: 10.1016/j.indcrop.2022.115684.
[2] N. A. M. Liyakathali, P. D. Muley, G. Aita, and D. Boldor, “Effect of frequency and reaction time in focused ultrasonic pretreatment of energy cane bagasse for bioethanol production,” Bioresource Technology, vol. 200, pp. 262–271, Jan. 2016, doi: 10.1016/j.biortech.2015.10.028.
[3] E. Olguin-Maciel, I. A. Jimenez-Villarreal, T. Toledano-Thompson, L. Alzate-Gaviria, and R. Tapia-Tussell, “Effect of ultrasound pretreatment combined with enzymatic hydrolysis of non-conventional starch on sugar yields to bioethanol conversion,” Biomass Conversion and Biorefinery, vol. 14, pp. 11469–11477, May 2024, doi: 10.1007/s13399-022-03217-2.
[4] Y. W. Sitotaw, N. G. Habtu, A. Y. Gebreyohannes, and T. Van Gerven, “Effect of alkaline combined with ultrasonic pretreatment to enhance enzymatic saccharification of sugar cane bagasse and Enset fiber (Ensete ventricosum),” Biomass Conversion and Biorefinery, vol. 14, pp. 3897–3915, Feb. 2024, doi: 10.1007/s13399-022-03021-y.
[5] V. Z. Ong and T. Y. Wu, “An application of ultrasonication in lignocellulosic biomass valorisation into bio-energy and bio-based products,” Renewable and Sustainable Energy Reviews, vol. 132, Oct. 2020, Art. no. 109924, doi: 10.1016/j.rser.2020.109924.
[6] K. M. Lee, M. F. Zanil, K. K. Chan, Z. P. Chin, Y. C. Liu, and S. Lim, “Synergistic ultrasound-assisted organosolv pretreatment of oil palm empty fruit bunches for enhanced enzymatic saccharification: An optimization study using artificial neural networks,” Biomass and Bioenergy, vol. 139, Aug. 2020, Art. no. 105621, doi: 10.1016/j.biombioe.2020.105621.
[7] K. Olofsson, M. Bertilsson, and G. Lidén, “A short review on SSF – an interesting process option for ethanol production from lignocellulosic feedstocks,” Biotechnology for Biofuels, vol. 1, 2008, Art. no. 7, doi: 10.1186/1754-6834-1-7.
[8] M. Ask et al., “Challenges in enzymatic hydrolysis and fermentation of pretreated Arundo donax revealed by a comparison between SHF and SSF,” Process Biochemistry, vol. 47, no. 10, pp. 1452–1459, Oct. 2012, doi: 10.1016/j.procbio.2012.05.016.
[9] D. Dahnum, S. O. Tasum, E. Triwahyuni, M. Nurdin, and H. Abimanyu, “Comparison of SHF and SSF processes using enzyme and dry yeast for optimization of bioethanol production from empty fruit bunch,” Energy Procedia, vol. 68, pp. 107–116, Apr. 2015, doi: 10.1016/j.egypro.2015.03.238.
[10] H. Zabed, J. N. Sahu, A. Suely, A. N. Boyce, and G. Faruq, “Bioethanol production from renewable sources: Current perspectives and technological progress,” Renewable and Sustainable Energy Reviews, vol. 71, pp. 475–501, May 2017, doi: 10.1016/j.rser.2016.12.076.
[11] R. Wu, Y. Li, X. Wang, Y. Fu, M. Qin, and Y. Zhang, “In-situ lignin sulfonation for enhancing enzymatic hydrolysis of poplar using mild organic solvent pretreatment,” Bioresource Technology, vol. 369, Art. no. 128410, Feb. 2023, doi: 10.1016/j.biortech.2022.128410.
[12] K. Stenberg, M. Bollók, K. Réczey, M. Galbe, and G. Zacchi, “Effect of substrate and cellulase concentration on simultaneous saccharification and fermentation of steam-pretreated softwood for ethanol production,” Biotechnology and Bioengineering, vol. 68, no. 2, pp. 204–210, Apr. 2000, doi: 10.1002/(SICI)1097-0290(20000420)68:23.0.CO;2-4.
[13] G. L. Miller, “Use of dinitrosalicylic acid reagent for determination of reducing sugar,” Analytical Chemistry, vol. 31, no. 3, pp. 426–428, Mar. 1959, doi: 10.1021/ac60147a030.
[14] A. Kaoloun, P. Tantayotai, L. Tabtimmai, N. Kottam, and A. Tawai, “Enhancing enzymatic hydrolysis of sugarcane leaves through sulfonation-based pretreatment with a reusable organic solvent under mild conditions for bioethanol production,” Applied Science and Engineering Progress, vol. 19, no. 1, 2026, Art. no. 7831, doi: 10.14416/j.asep.2025.06.009.
[15] R. Poludasu, B. Piyarat, and C. Benjamas, “Optimized ultrasound-assisted peracetic acid pretreatment approach for enhanced sugars and decreased inhibitors for bio-ethanol production from OPEFB,” Vietnam Journal of Science and Technology, vol. 62, no. 6, pp. 1159–1172, 2024, doi: 10.15625/2525-2518/18645.
[16] C. Panjapornpon, A. Kaoloun, M. Sriariyanun, K. Katam, S. K. Maity, and A. Tawai, “Simulation-based process design and preliminary techno-economic assessment of sulfonation-based pretreatment of sugarcane leaves under mild conditions,” Waste and Biomass Valorization, vol. 17, pp. 5107–5121, Sep. 2026, doi: 10.1007/s12649-025-03428-4.
[17] X. Zhuang, Y. Liu, Z. Wu, Y. Feng, X. Pan, and H. Qiao, “Ultrasonic-formic acid pretreatment coupled with metal ion/deep eutectic synergistic catalysis: Efficient conversion of biomass to 5-hydroxymethylfurfural,” Polymers, vol. 18, no. 2, 2026, Art. no. 218, doi: 10.3390/polym18020218.
[18] P. Pathak, A. Gupta, N. K. Bhardwaj, A. Goyal, and V. S. Moholkar, “Impact of mild and harsh conditions of formic acid-based organosolv pretreatment on biomass fractionation of sugarcane tops,” Biomass Conversion and Biorefinery, vol. 11, pp. 2027–2040, Oct. 2021, doi: 10.1007/s13399-020-00629-w.
[19] A. A. Schmatz, A. M. Salazar-Bryam, J. Contiero, C. Sant’Anna, and M. Brienzo, “Pseudo-lignin content decreased with hemicellulose and lignin removal, improving cellulose accessibility, and enzymatic digestibility,” BioEnergy Research, vol. 14, pp. 106–121, Mar. 2021, doi: 10.1007/s12155-020-10187-8.
[20] K. Öhgren, R. Bura, G. Lesnicki, J. Saddler, and G. Zacchi, “A comparison between simultaneous saccharification and fermentation and separate hydrolysis and fermentation using steam-pretreated corn stover,” Process Biochemistry, vol. 42, no. 5, pp. 834–839, May 2007, doi: 10.1016/j.procbio.2007.02.003.
DOI: 10.14416/j.asep.2026.11.005
Refbacks
- There are currently no refbacks.
Applied Science and Engineering Progress






