Optimization of Process Conditions for Production of Succinic Acid and Lactic Acids from Cake Waste Using Response Surface Methodology
DOI:
https://doi.org/10.52151/jae2026632.2008Keywords:
Box-Behnken design, food waste valorization, microbial fermentation, statistical modeling, sustainable bio-processingAbstract
Food waste valorization into bio-based chemicals is an eco-friendly strategy to address environmental pollution and reduce reliance on fossil-based resources. This study explores the feasibility of bioconversion of cake waste into succinic acid (SA) and lactic acid (LA), through enzymatic hydrolysis and microbial fermentation. The compositional analysis revealed that, on a dry weight basis, cake waste contains 572.0 mg g⁻¹ carbohydrates, 495.0 mg g⁻¹ starch, and 69.3 mg g⁻¹ protein, making it a nutritionally rich substrate for fermentation and then it was hydrolyzed using α-amylase, glucoamylase, and protease to release fermentable sugars and amino nitrogen. Enzymatic hydrolysis yielded 41.50 g L⁻¹ glucose and 4.56 g L⁻¹ free amino nitrogen. The enzymatic hydrolysate was subsequently fermented using Escherichia coli and Bacillus coagulans for SA and LA production, respectively. High performance liquid chromatography (HPLC) was used to quantify organic acid concentrations. Process optimization was performed using Box Behnken Design under response surface methodology (RSM), with three independent variables, namely, temperature, pH, and incubation time. The highest succinic acid yield (0.2194 g g⁻¹ cake waste) was obtained at a temperature of 40°C, pH of 7.5, and an incubation time of 144 h, while the maximum lactic acid yield (0.3178 g g⁻¹ cake waste) was recorded at a temperature of 47°C, pH of 5.6, and an incubation time of 94 h. Results demonstrate that cake waste can serve as a promising low-cost substrate for sustainable production of succinic and lactic acids, highlighting its potential for efficient food waste valorization through microbial bioprocessing.
Downloads
References
Abdel-Rahman, M. A., Tashiro, Y., & Sonomoto, K. (2013). Recent advances in lactic acid production by microbial fermentation processes. Biotechnology Advances, 31(6), 877–902. https://doi.org/10.1016/j.biotechadv.2013.04.002 DOI: https://doi.org/10.1016/j.biotechadv.2013.04.002
Ahn, J. H., Seo, H., Park, W., Seok, J., Lee, J. A., Kim, W. J., Kim, G. B., Kim, K.-J., & Lee, S. Y. (2020). Enhanced succinic acid production by Mannheimia employing optimal malate dehydrogenase. Nature Communications, 11(1), 1970. https://doi.org/10.1038/s41467-020-15839-z DOI: https://doi.org/10.1038/s41467-020-15839-z
Alam, M. S., Khaira, H., Pathania, S., Kumar, S. & Singh, B. (2015). Extrusion process optimization for soy-carrot pomace powder incorporated wheat-based snacks. Journal of Agricultural Engineering (India), 52(1), 1–13. https://doi.org/10.52151/jae2015521.1565 DOI: https://doi.org/10.52151/jae2015521.1565
Alexandri, M., Blanco-Catalá, J., Schneider, R., Turon, X., & Venus, J. (2020). High L(+)-lactic acid productivity in continuous fermentations using bakery waste and lucerne green juice as renewable substrates. Bioresource Technology, 316, 123949. https://doi.org/10.1016/j.biortech.2020.123949 DOI: https://doi.org/10.1016/j.biortech.2020.123949
Ali, S., Narwari, I. S., & Faisal, S. (2025). Response Surface Optimization of Refractance Window Drying of Bitter Gourd Slices. Journal of Agricultural Engineering (India), 61(1), 52–66. https://doi.org/10.52151/jae2024611.1829 DOI: https://doi.org/10.52151/jae2024611.1829
Arancon, R. A. D., Lin, C. S. K., Chan, K. M., Kwan, T. H., & Luque, R. (2013). Advances on waste valorization: new horizons for a more sustainable society. Energy Science & Engineering, 1(2), 53–71. https://doi.org/10.1002/ese3.9 DOI: https://doi.org/10.1002/ese3.9
Becker, J., Lange, A., Fabarius, J., & Wittmann, C. (2015). Top value platform chemicals: bio-based production of organic acids. Current Opinion in Biotechnology, 36, 168–175. https://doi.org/10.1016/j.copbio.2015.08.022 DOI: https://doi.org/10.1016/j.copbio.2015.08.022
Behera, S. K., Meena, H., Chakraborty, S., & Meikap, B. C. (2018). Application of response surface methodology (RSM) for optimization of leaching parameters for ash reduction from low-grade coal. International Journal of Mining Science and Technology, 28(4), 621–629. https://doi.org/10.1016/j.ijmst.2018.04.014 DOI: https://doi.org/10.1016/j.ijmst.2018.04.014
Bibwe, B., Mani, I., Kar, A., Samuel, D. V. K., & Iquebal, M. A. (2024). Optimization of oil loading and starch-protein ratio for encapsulation of flaxseed oil using response surface methodology. Journal of Agricultural Engineering (India), 56(2), 80–90. https://doi.org/10.52151/jae2018551.1680 DOI: https://doi.org/10.52151/jae2018551.1680
Borham, A., Haroun, M., Saleh, I. A., Zomot, N., Okla, M. K., Askar, M., Elmasry, M., Elshahat, A., Liu, L., Zhao, C., Wang, J., & Qian, X. (2024). A statistical optimization for almost-complete methylene blue biosorption by Gracilaria bursa-pastoris. Heliyon, 10(15), e34972. https://doi.org/10.1016/j.heliyon.2024.e34972 DOI: https://doi.org/10.1016/j.heliyon.2024.e34972
Chikkanna, G. S., Samuel, D. V. K., & Jha, S. K. (2015). Optimization of extrusion process for preparation of ready-to-eat product from maize-rice-anola. Journal of Agricultural Engineering (India), 52(2), 28–36. https://doi.org/10.52151/jae2015522.1575 DOI: https://doi.org/10.52151/jae2015522.1575
Coma, M., Martinez-Hernandez, E., Abeln, F., Raikova, S., Donnelly, J., Arnot, T. C., Allen, M. J., Hong, D. D., & Chuck, C. J. (2017). Organic waste as a sustainable feedstock for platform chemicals. Faraday Discussions, 202, 175–195. https://doi.org/10.1039/C7FD00070G DOI: https://doi.org/10.1039/C7FD00070G
Du, C., Lin, S. K. C., Koutinas, A., Wang, R., Dorado, P., & Webb, C. (2008). A wheat biorefining strategy based on solid-state fermentation for fermentative production of succinic acid. Bioresource Technology, 99(17), 8310–8315. https://doi.org/10.1016/j.biortech.2008.03.019 DOI: https://doi.org/10.1016/j.biortech.2008.03.019
Fard Masoumi, H. R., Basri, M., Kassim, A., Kuang Abdullah, D., Abdollahi, Y., Abd Gani, S. S., & Rezaee, M. (2013). Statistical Optimization of Process Parameters for Lipase‐Catalyzed Synthesis of Triethanolamine‐Based Esterquats Using Response Surface Methodology in 2‐Liter Bioreactor. The Scientific World Journal, 2013(1). https://doi.org/10.1155/2013/962083 DOI: https://doi.org/10.1155/2013/962083
Faulks, R. M., & Timms, S. B. (1985). A rapid method for determining the carbohydrate component of dietary fibre. Food Chemistry, 17(4), 273–287. https://doi.org/10.1016/0308-8146(85)90036-6 DOI: https://doi.org/10.1016/0308-8146(85)90036-6
Förster, A. H., & Gescher, J. (2014). Metabolic Engineering of Escherichia coli for Production of Mixed-Acid Fermentation End Products. Frontiers in Bioengineering and Biotechnology, 2. https://doi.org/10.3389/fbioe.2014.00016 DOI: https://doi.org/10.3389/fbioe.2014.00016
Han, W., Xu, X., Gao, Y., He, H., Chen, L., Tian, X., & Hou, P. (2019). Utilization of waste cake for fermentative ethanol production. Science of The Total Environment, 673, 378–383. https://doi.org/10.1016/j.scitotenv.2019.04.079 DOI: https://doi.org/10.1016/j.scitotenv.2019.04.079
Hiramoto, K., Kubo, S., Tsuji, K., Sugiyama, D., Iizuka, Y., & Hamano, H. (2023). Bacillus coagulans (species of lactic acid-forming Bacillus bacteria) ameliorates azoxymethane and dextran sodium sulfate-induced colon cancer in mice. Journal of Functional Foods, 100, 105406. https://doi.org/10.1016/j.jff.2023.105406 DOI: https://doi.org/10.1016/j.jff.2023.105406
Hodge, J. E., & Hofreiter, B. T. (1962). Determination of reducing sugars and carbohydrates. In: R. L. Whistler, & M. L. Wolfrom (Eds.), Methods in carbohydrate chemistry (pp. 380-394). Academic Press, New York.
Jantama, K., Zhang, X., Moore, J. C., Shanmugam, K. T., Svoronos, S. A., & Ingram, L. O. (2008). Eliminating side products and increasing succinate yields in engineered strains of Escherichia coli C. Biotechnology and Bioengineering, 101(5), 881–893. https://doi.org/10.1002/bit.22005 DOI: https://doi.org/10.1002/bit.22005
Jiang, S., Xu, P., & Tao, F. (2019). l-Lactic acid production by Bacillus coagulans through simultaneous saccharification and fermentation of lignocellulosic corncob residue. Bioresource Technology Reports, 6, 131–137. https://doi.org/10.1016/j.biteb.2019.02.005 DOI: https://doi.org/10.1016/j.biteb.2019.02.005
Lee, J., Chen, W.-H., & Park, Y.-K. (2022). Recent achievements in platform chemical production from food waste. Bioresource Technology, 366, 128204. https://doi.org/10.1016/j.biortech.2022.128204 DOI: https://doi.org/10.1016/j.biortech.2022.128204
Lee, S. J., Lee, D.-Y., Kim, T. Y., Kim, B. H., Lee, J., & Lee, S. Y. (2005). Metabolic engineering of Escherichia coli for enhanced production of succinic acid, based on genome comparison and in silico gene knockout simulation. Applied and Environmental Microbiology, 71(12), 7880–7887. https://doi.org/10.1128/AEM.71.12.7880-7887.2005 DOI: https://doi.org/10.1128/AEM.71.12.7880-7887.2005
Lin, C. S. K., Koutinas, A. A., Stamatelatou, K., Mubofu, E. B., Matharu, A. S., Kopsahelis, N., Pfaltzgraff, L. A., Clark, J. H., Papanikolaou, S., Kwan, T. H., & Luque, R. (2014). Current and future trends in food waste valorization for the production of chemicals, materials and fuels: a global perspective. Biofuels, Bioproducts and Biorefining, 8(5), 686–715. https://doi.org/10.1002/bbb.1506 DOI: https://doi.org/10.1002/bbb.1506
Liu, Z., & Pan, J. (2017). A practical method for extending the biuret assay to protein determination of corn-based products. Food Chemistry, 224, 289–293. https://doi.org/10.1016/j.foodchem.2016.12.084 DOI: https://doi.org/10.1016/j.foodchem.2016.12.084
Loewus, F. A. (1952). Improvement in Anthrone Method for Determination of Carbohydrates. Analytical Chemistry, 24(1), 219–219. https://doi.org/10.1021/ac60061a050 DOI: https://doi.org/10.1021/ac60061a050
Ma, K., Hu, G., Pan, L., Wang, Z., Zhou, Y., Wang, Y., Ruan, Z., & He, M. (2016). Highly efficient production of optically pure l-lactic acid from corn stover hydrolysate by thermophilic Bacillus coagulans. Bioresource Technology, 219, 114–122. https://doi.org/10.1016/j.biortech.2016.07.100 DOI: https://doi.org/10.1016/j.biortech.2016.07.100
Manmai, N., Unpaprom, Y., & Ramaraj, R. (2021). Bioethanol production from sunflower stalk: application of chemical and biological pretreatments by response surface methodology (RSM). Biomass Conversion and Biorefinery, 11(5), 1759–1773. https://doi.org/10.1007/s13399-020-00602-7 DOI: https://doi.org/10.1007/s13399-020-00602-7
Marsden, W. L., Gray, P. P., Nippard, G. J., & Quinlan, M. R. (1982). Evaluation of the DNS method for analysing lignocellulosic hydrolysates. Journal of Chemical Technology and Biotechnology, 32(7–12), 1016–1022. https://doi.org/10.1002/jctb.5030320744 DOI: https://doi.org/10.1002/jctb.5030320744
Misiou, O., Zourou, C., & Koutsoumanis, K. (2021). Development and validation of a predictive model for the effect of temperature, pH and water activity on the growth kinetics of Bacillus coagulans in non-refrigerated ready-to-eat food products. Food Research International, 149, 110705. https://doi.org/10.1016/j.foodres.2021.110705 DOI: https://doi.org/10.1016/j.foodres.2021.110705
Onyeaka, H., Mansa, R. F., Wong, C. M. V. L., & Miri, T. (2022). Bioconversion of starch base food waste into bioethanol. Sustainability, 14(18), 11401. https://doi.org/10.3390/su141811401 DOI: https://doi.org/10.3390/su141811401
Packiyadhas, P., Sivaperumal, S. K., & Murugesan, S. (2025). A comprehensive review of food waste: Composition, current management, thermal treatment, valorization into bioproducts and sustainable development goals linkages. Journal of Material Cycles and Waste Management, 27, 777–795. https://doi.org/10.1007/s10163-024-02153-9 DOI: https://doi.org/10.1007/s10163-024-02153-9
Parekh, S., Vinci, V. A., & Strobel, R. J. (2000). Improvement of microbial strains and fermentation processes. Applied Microbiology and Biotechnology, 54(3), 287–301. https://doi.org/10.1007/s002530000403 DOI: https://doi.org/10.1007/s002530000403
Pleissner, D., Demichelis, F., Mariano, S., Fiore, S., Navarro Gutiérrez, I. M., Schneider, R., & Venus, J. (2017). Direct production of lactic acid based on simultaneous saccharification and fermentation of mixed restaurant food waste. Journal of Cleaner Production, 143, 615–623. https://doi.org/10.1016/j.jclepro.2016.12.065 DOI: https://doi.org/10.1016/j.jclepro.2016.12.065
Raghuramulu, N., Nair, K. M., & Kalyanasundaram, S. (1983). A manual of laboratory techniques. National Institute of Nutrition, Indian Council of Medical Research, Hyderabad, India.
Rahimi-Gorji, M., Pourmehran, O., Hatami, M., & Ganji, D. D. (2015). Statistical optimization of microchannel heat sink (MCHS) geometry cooled by different nanofluids using RSM analysis. The European Physical Journal Plus, 130(2), 22. https://doi.org/10.1140/epjp/i2015-15022-8 DOI: https://doi.org/10.1140/epjp/i2015-15022-8
Rastogi, A., & Banerjee, R. (2020). Statistical optimization of bacterial cellulose production by Leifsonia soli and its physico-chemical characterization. Process Biochemistry, 91, 297–302. https://doi.org/10.1016/j.procbio.2019.12.021 DOI: https://doi.org/10.1016/j.procbio.2019.12.021
Sivarajasekar, N., Balasubramani, K., Mohanraj, N., Prakash Maran, J., Sivamani, S., Ajmal Koya, P., & Karthik, V. (2017). Fixed-bed adsorption of atrazine onto microwave irradiated Aegle marmelos Correa fruit shell: Statistical optimization, process design and breakthrough modeling. Journal of Molecular Liquids, 241, 823–830. https://doi.org/10.1016/j.molliq.2017.06.064 DOI: https://doi.org/10.1016/j.molliq.2017.06.064
Solzin, J., Eppler, K., Knapp, B., Buchner, H., & Bluhmki, E. (2023). Optimising cell-based bioassays via integrated design of experiments (ixDoE) - A practical guide. SLAS Discovery, 28(1), 29–38. https://doi.org/10.1016/j.slasd.2022.10.004 DOI: https://doi.org/10.1016/j.slasd.2022.10.004
Song, H., & Lee, S. Y. (2006). Production of succinic acid by bacterial fermentation. Enzyme and Microbial Technology, 39(3), 352–361. https://doi.org/10.1016/j.enzmictec.2005.11.043 DOI: https://doi.org/10.1016/j.enzmictec.2005.11.043
Stat-Ease. (2021). Design-Expert® software (Version 13) [Computer software]. Stat-Ease, Inc. https://www.statease.com
Sun, Z., Li, M., Qi, Q., Gao, C., & Lin, C. S. K. (2014). Mixed Food Waste as Renewable Feedstock in Succinic Acid Fermentation. Applied Biochemistry and Biotechnology, 174(5), 1822–1833. https://doi.org/10.1007/s12010-014-1169-7 DOI: https://doi.org/10.1007/s12010-014-1169-7
Suresh, T., Sivarajasekar, N., Balasubramani, K., Ahamad, T., Alam, M., & Naushad, M. (2020). Process intensification and comparison of bioethanol production from food industry waste (potatoes) by ultrasonic assisted acid hydrolysis and enzymatic hydrolysis: Statistical modelling and optimization. Biomass and Bioenergy, 142, 105752. https://doi.org/10.1016/j.biombioe.2020.105752 DOI: https://doi.org/10.1016/j.biombioe.2020.105752
Uçkun Kiran, E., Trzcinski, A. P., Ng, W. J., & Liu, Y. (2014). Bioconversion of food waste to energy: A review. Fuel, 134, 389–399. https://doi.org/10.1016/j.fuel.2014.05.074 DOI: https://doi.org/10.1016/j.fuel.2014.05.074
Ude, M. U., Oluka, I., & Eze, P. C. (2020). Optimization and kinetics of glucose production via enzymatic hydrolysis of mixed peels. Journal of Bioresources and Bioproducts, 5(4), 283–290. https://doi.org/10.1016/j.jobab.2020.10.007 DOI: https://doi.org/10.1016/j.jobab.2020.10.007
Vera Zambrano, M., Dutta, B., Mercer, D. G., MacLean, H. L., & Touchie, M. F. (2019). Assessment of moisture content measurement methods of dried food products in small-scale operations in developing countries: A review. Trends in Food Science & Technology, 88, 484–496. https://doi.org/10.1016/j.tifs.2019.04.006 DOI: https://doi.org/10.1016/j.tifs.2019.04.006
Vohra, A., & Satyanarayana, T. (2002). Statistical optimization of the medium components by response surface methodology to enhance phytase production by Pichia anomala. Process Biochemistry, 37(9), 999–1004. https://doi.org/10.1016/S0032-9592(01)00308-9 DOI: https://doi.org/10.1016/S0032-9592(01)00308-9
Volkmar, M., Maus, A.-L., Weisbrodt, M., Bohlender, J., Langsdorf, A., Holtmann, D., & Ulber, R. (2023). Municipal green waste as substrate for the microbial production of platform chemicals. Bioresources and Bioprocessing, 10(1), 43. https://doi.org/10.1186/s40643-023-00663-2 DOI: https://doi.org/10.1186/s40643-023-00663-2
Wang, F., Zhou, C., He, W., Zhu, H., Huang, J., & Li, G. (2016). The content variation of fat, protein and starch in kitchen waste under microwave radiation. Procedia Environmental Sciences, 31, 530–534. https://doi.org/10.1016/j.proenv.2016.02.075 DOI: https://doi.org/10.1016/j.proenv.2016.02.075
Wheeldon, I., Christopher, P., & Blanch, H. (2017). Integration of heterogeneous and biochemical catalysis for production of fuels and chemicals from biomass. Current Opinion in Biotechnology, 45, 127–135. https://doi.org/10.1016/j.copbio.2017.02.019 DOI: https://doi.org/10.1016/j.copbio.2017.02.019
Zhang, A. Y., Sun, Z., Leung, C. C. J., Han, W., Lau, K. Y., Li, M., & Lin, C. S. K. (2013). Valorisation of bakery waste for succinic acid production. Green Chemistry, 15(3), 690. https://doi.org/10.1039/c2gc36518a DOI: https://doi.org/10.1039/c2gc36518a
