Production and optimisation of bacterial cellulose from banana (musa spp.) banana rachis juice pre-treated with ozone using a box–behnken design

Authors

DOI:

https://doi.org/10.26439/ing.ind2026.n50.8611

Keywords:

bacterial cellulose, banana rachis, ozone, optimization, response surface, scale-up

Abstract

The valorisation of banana rachis (BR) is a key strategy within the circular bioeconomy of tropical regions. This study evaluated the production of bacterial cellulose (BC) by Komagataeibacter hansenii ATCC 23769 using banana rachis juice (BRJ) pretreated with ozone (600 mg O2 h-¹). Through a Box-Behnken design, the BRJ volume fraction (25-75 % v/v) and fermentation time (7-21 days) were identified as the critical factors affecting yield, whilst ozonation time (10-30 min) exhibited a limited effect. Under optimal conditions (75 % v/v BRJ, 21 days of fermentation), a maximum yield of 6,00 ± 0,56 g L-¹ was achieved, with a response surface model exhibiting R² = 0,7894. Scale-up to a 20 L static bioreactor confirmed the robustness of the system, maintaining approximately 6 g L-¹. Characterisation via FTIR-ATR, XRD, and TGA confirmed type I cellulose of high structural purity. These findings position ozone-pretreated BRJ as a sustainable substrate for BC production, thereby contributing to the valorisation of agro-industrial waste.

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Author Biographies

  • Andres Joel Zeas Sesme, Facultad de Posgrado, Universidad Estatal de Milagro, Ecuador

    Ingeniero en Biotecnología por la Universidad Estatal de Milagro en Ecuador, institución donde actualmente cursa su maestría en Biotecnología. Su trayectoria profesional incluye el desempeño como asistente técnico de investigación y asistente de laboratorio en la misma casa de estudios, donde participó en proyectos enfocados en la bioeconomía circular y el diseño experimental de bioprocesos. En la actualidad, ejerce como docente en la Unidad Educativa Albert Einstein en Milagro, Ecuador. Su labor científica se especializa en la optimización de la producción de celulosa bacteriana a partir de jugo de raquis de banano mediante pretratamiento con ozono, utilizando modelos avanzados como ANN y RSM.

  • Thaily Jazmin Martínez Castillo , Facultad de Posgrado, Universidad Estatal de Milagro, Ecuador

    Ingeniera en Biotecnología por la Universidad Estatal de Milagro en Ecuador, donde actualmente cursa la maestría en Biotecnología. Su formación académica se ha orientado a la aplicación de la biotecnología en sistemas productivos, con énfasis en el control de calidad y el aseguramiento de procesos. Actualmente, se desempeña en el Área de Control de Calidad de la empresa camaronera Tropack, donde es responsable de la verificación, monitoreo y cumplimiento de los estándares aplicables a los procesos productivos, garantizando la inocuidad y la calidad del producto final.

  • Manuel Alejandro Fiallos Cárdenas, Facultad de Salud y Servicios Sociales, Universidad Estatal de Milagro, Ecuador

    Ingeniero químico por la Universidad de Guayaquil y biotecnólogo por la Escuela Superior Politécnica del Litoral (ESPOL), especializado en la transformación de residuos agroindustriales en bioproductos de alto valor. Como profesor e investigador en la Universidad Ikiam, lidera iniciativas de sostenibilidad y bioeconomía circular en la Amazonía. Es un experto reconocido en la producción de nanocelulosa bacteriana y su aplicación en soluciones de empaque ecológicas, donde integra análisis técnicos, económicos y ambientales

References

Abdel Hakim, A., & Mourad, R. (2023). Nanocellulose and its polymer composites: Preparation, characterization, and applications. Russian Chemical Reviews, 92(4). https://doi.org/10.57634/RCR5076

Abdoussalami, A., Hu, Z., Islam, A. R. M. T., & Wu, Z. (2023). Climate change and its impacts on banana production: A systematic analysis. Environment, Development and Sustainability, 25(11), 12217-12246. https://doi.org/10.1007/s10668-023-03168-2

Al-Baarri, A. N., Legowo, A. M., Abduh, S. B. M., Mawarid, A. A., Farizha, K. M., & Silvia, M. (2019). Production of ozone and the simple detection using potassium iodide titration method. IOP Conference Series: Earth and Environmental Science, 292(1), 12062. https://doi.org/10.1088/1755-1315/292/1/012062

Almeida, D. M., Aparecida Prestes, R., Fonseca, A. F. da, Woiciechowski, A. L., & Wosiacki, G.

(2013). Minerals consumption by Acetobacter xylinum on cultivation medium on coconut water. Brazilian Journal of Microbiology, 44(1), 197-206. https://doi.org/10.1590/S1517-83822013005000012

Alzate Acevedo, S., Díaz Carrillo, Á. J., Flórez-López, E., & Grande-Tovar, C. D. (2021). Recovery of banana waste-loss from production and processing: A contribution to a circular economy. Molecules, 26(17), 5282. https://doi.org/10.3390/molecules26175282

Arias-Roblero, M., Mora-Villalobos, V., & Velazquez-Carrillo, C. (2021). Evaluation of fed-batch fermentation for production of polyhydroxybutyrate with a banana pulp juice substrate from an agro industrial by-product. Frontiers in Sustainable Food Systems, 5. https://doi.org/10.3389/fsufs.2021.681596

Barbadillo Jove, F. (2015). Estudio cinético de degradación térmica de poliuretanos mediante análisis termogravimétrico (TGA) [Tesis doctoral, Universidad de La Coruña]. Repositorio Instucional da Universidade da Coruña. http://hdl.handle.net/2183/14521

Bhattacharya, A., Sadaf, A., Dubey, S., Singh, R. P., & Khare, S. K. (2021). Production and characterization of Komagataeibacter xylinus SGP8 nanocellulose and its calcite based composite for removal of Cd ions. Environmental Science and Pollution Research, 28, 46423-46430. https://doi.org/10.1007/s11356-020-08845-7

Bonifazi, G., Gasbarrone, R., & Serranti, S. (2023). Evaluation of sugar content in hopped wort of artisanal beer by shortwave infrared spectroscopy. En M. S. Kim & B.-K. Cho (Eds.), Sensing for agriculture and food quality and safety XV. SPIE Defense + Commercial Sensing. https://doi.org/10.1117/12.2663382

Borsoi, C., Zimmernnam, M. V. G., Zattera, A. J., Santana, R. M. C., & Ferreira, C. A. (2016). Thermal degradation behavior of cellulose nanofibers and nanowhiskers. Journal of Thermal Analysis and Calorimetry, 126, 1867-1878. https://doi.org/10.1007/s10973-016-5653-x

Burgos Montañez, L. J. (2020). Cuantificación de azúcares reductores del sustrato en residuos de piña con el método del ácido 3,5-dinitrosalicílico. Revista de Investigación, 13(1), 57-66. https://doi.org/10.29097/23461098.308

Choudhary, A., Kumar, A., Kandpal, R., Gupta, A. K., Jha, A. K., Naik, B., Kumar, V., Rustagi, S., Chutia, H., & Khan, J. M. (2024). Evaluation of secondary metabolites, nutraceutical potential and amino acid profile of fresh dates (Phoenix dactylifera) alcoholic beverage. Discover Food, 4, Artículo 53. https://doi.org/10.1007/s44187-024-00137-0

Deumaga, M. F. T., Emaga, T. H., Tchokouassom, R., Vanderghem, C., Aguedo, M., Gillet, S., Jacquet, N., Danthine, S., Magali, D., & Richel, A. (2015). Genotype contribution to the chemical composition of banana rachis and implications for thermo/biochemical conversion. Biomass Conversion and Biorefinery, 5, 409-416. https://doi.org/10.1007/s13399-015-0158-6

Fiallos-Cardenas, M., Gavin, C., Huilcarema-Enríquez, K., Cumbicus-Bravo, A., & Pozo, F. (2025). Innovation in obtaining bacterial nanocellulose from banana rachis: Effects of ozone treatment. Case Studies in Chemical and Environmental Engineering, 11, 101044. https://doi.org/10.1016/j.cscee.2024.101044

Fiallos-Cárdenas, M., Pérez-Martínez, S., & Ramirez, A. D. (2022). Prospectives for the development of a circular bioeconomy around the banana value chain. Sustainable Production and Consumption, 30, 541-555. https://doi.org/10.1016/j.spc.2021.12.014

Fiallos-Cárdenas, M., Ramirez, A. D., Pérez-Martínez, S., Bonilla, H. R., Ordoñez-Viñan, M., Ruiz-Barzola, O., & Reinoso, M. A. (2021). Bacterial nanocellulose derived from banana leaf extract: Yield and variation factors. Resources, 10(12), 121. https://doi.org/10.3390/resources10120121

García-Cubero, M. T., González-Benito, G., Indacoechea, I., Coca, M., & Bolado, S. (2009). Effect of ozonolysis pretreatment on enzymatic digestibility of wheat and rye straw. Bioresource Technology, 100(4), 1608-1613. https://doi.org/10.1016/j.biortech.2008.09.012

Guevara, K. M., Martínez-Valenzuela, G., Sánchez-Vásquez, V., Guerrero-Ruiz, K., & Fiallos-Cárdenas, M. (2025). Trends and perspectives on bacterial nanocellulose: A comprehensive analysis from the three helixes of innovation. Materials Today Sustainability, 30, Artículo 101090. https://doi.org/10.1016/j.mtsust.2025.101090

Huang, C., Guo, H.-J., Xiong, L., Wang, B., Shi, S.-L., Chen, X.-F., Lin, X.-Q., Wang, C., Luo, J., & Chen, X.-D. (2016). Using wastewater after lipid fermentation as substrate for bacterial cellulose production by Gluconacetobacter xylinus. Carbohydrate Polymers, 136, 198-202. https://doi.org/10.1016/j.carbpol.2015.09.043

Irving, D., Bakhshandeh, S., Tran, T. K. A., & McBratney, A. B. (2024). A cost-effective method for quantifying soil respiration. Soil Security, 16, 100162. https://doi.org/10.1016/j.soisec.2024.100162

Jahed, E., Khodaparast, M. H. H., & Mousavi Khaneghah, A. (2014). Bentonite, temperature and pH effects on purification indexes of raw sugar beet juice to production of inverted liquid sugar. Applied Clay Science, 102, 155-163. https://doi.org/10.1016/j.clay.2014.09.036

Jeridi, M., Siddiqui, S., Siddiqua, A., Moneim, D. A., Aika, E. A. M., Zahrani, F., Essenidi, M., & Ferchichi, A. (2023). Nutritional analysis of fresh banana fruits (Musa spp.) grown in South Tunisia. Bangladesh Journal of Botany, 52(2), 253-260. https://doi.org/10.3329/bjb.v52i2.67010

Kumari, P., Ravi Kiran, B., & Venkata Mohan, S. (2022). Polyhydroxybutyrate production by Chlorella sorokiniana SVMIICT8 under nutrient-deprived mixotrophy. Bioresource Technology, 354, Artículo 127135. https://doi.org/10.1016/j.biortech.2022.127135

Lima, H. L. S., Nascimento, E. S., Andrade, F. K., Brígida, A. I. S., Borges, M. F., Cassales, A. R., Muniz, C. R., Souza Filho, M. D. S. M., Morais, J. P. S., & Rosa, M. D. F. (2017). Bacterial cellulose production by Komagataeibacter hansenii ATCC 23769 using sisal juice—An agroindustry waste. Brazilian Journal of Chemical Engineering, 34(3), 671-680. https://doi.org/10.1590/0104-6632.20170343s20150514

Martínez-Solórzano, G. E., & Rey-Brina, J. C. (2021). Bananos (Musa AAA): importancia, producción y comercio en tiempos de Covid-19. Agronomía Mesoamericana, 32(3), 1034-1046. https://doi.org/10.15517/am.v32i3.43610

Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31(3), 426-428. https://doi.org/10.1021/ac60147a030

Minardi, C., Bersanetti, D., Sarlin, E., Santala, V., & Mangayil, R. (2024). Optimization of citrus pulp waste-based medium for improved bacterial nanocellulose production. Microorganisms, 12(10), 2095. https://doi.org/10.3390/microorganisms12102095

Mulyono, Nurbaiti, Rilyanti, M., & Herasari, D. (2024). Liquid pineapple waste as substrate for the production of bacterial nanocellulose (BNC) by local isolated microbe Kc-D-4. AIP Conference Proceedings, 2970(1), 60013. https://doi.org/10.1063/5.0210870

Muñoz S., K., Ponce G., M., Burgos B., G., Alcívar C., U., & Munizaga P., D. (2025). Capacidad antioxidante y composición fenólica en extractos de cerveza artesanal tipo ale. infoANALÍTICA, 13(1), 45-75. https://doi.org/10.26807/ia.v13i1.283

Nanda, S., Patra, B. R., Patel, R., Bakos, J., & Dalai, A. K. (2022). Innovations in applications and prospects of bioplastics and biopolymers: A review. Environmental Chemistry Letters, 20, 379-395. https://doi.org/10.1007/s10311-021-01334-4

Nascimento, R., Carvalheira, M., Crespo, J., & Neves, L. (2023). Extraction and characterization of cellulose obtained from banana plant pseudostem. Clean Technologies, 5(3), 1028-1043. https://doi.org/10.3390/cleantechnol5030052

Ono, Y., Takeuchi, M., Kimura, S., Puangsin, B., Wu, C.-N., & Isogai, A. (2022). Structures, molar mass distributions, and morphologies of TEMPO-oxidized bacterial cellulose fibrils. Cellulose, 29(9), 4977-4992. https://doi.org/10.1007/s10570-022-04617-3

Pacheco, G., Mello, C. V. de, Chiari-Andréo, B. G., Isaac, V. L. B., Ribeiro, S. J. L., Pecoraro, É., & Trovatti, E. (2018). Bacterial cellulose skin masks-properties and sensory tests. Journal of Cosmetic Dermatology, 17(5), 840-847. https://doi.org/10.1111/jocd.12441

Pereira, B., Li, Z.-J., De Mey, M., Lim, C. G., Zhang, H., Hoeltgen, C., & Stephanopoulos, G. (2016). Efficient utilization of pentoses for bioproduction of the renewable two-carbon compounds ethylene glycol and glycolate. Metabolic Engineering, 34, 80-87. https://doi.org/10.1016/j.ymben.2015.12.004

Peretz, R., Sterenzon, E., Gerchman, Y., Kumar Vadivel, V., Luxbacher, T., & Mamane, H. (2019). Nanocellulose production from recycled paper mill sludge using ozonation pretreatment followed by recyclable maleic acid hydrolysis. Carbohydrate Polymers, 216, 343-351. https://doi.org/10.1016/j.carbpol.2019.04.003

Pineda, L., Caicedo, L. M., & Riascos, C. (2012). Técnicas de fermentación y aplicaciones de la celulosa bacteriana: una revisión. Ingeniería y Ciencia, 8(16), 307-335. https://doi.org/10.17230/ingciencia.8.16.12

Ruiz-Molina, V. E., Soriano-Melgar, L. de A. A., Cortez-Mazatán, G. Y., Hernández-Zárate, G., Castillo-Zamudio, R. I., Flores-Estévez, N., Peralta-Rodríguez, R. D., & Noa-Carrazana, J. C. (2025). The potential effect of banana by-products on the in vitro inhibition of Lasiodiplodia theobromae and Colletotrichum sp. Journal of Agriculture and Food Research, 22, 102055. https://doi.org/10.1016/j.jafr.2025.102055

Saleh, A. K., El-Gendi, H., Ray, J. B., & Taha, T. H. (2021). A low-cost effective media from starch kitchen waste for bacterial cellulose production and its application as simultaneous absorbance for methylene blue dye removal. Biomass Conversion and Biorefinery, 13, 12437-12449. https://doi.org/10.1007/s13399-021-01973-1

Samanta, P., Senapati, T., Dey, S., & Ghosh, A. R. (2023). An overview of biomass conversion from agricultural waste: Address on environmental sustainability. En S. Singh, P. Singh, A. Sharma & M. Choudhury (Eds.), Agriculture Waste Management and Bioresource: The Circular Economy Perspective (pp. 46-77). Wiley. https://doi.org/10.1002/9781119808428.ch3

Segal, L., Creely, J. J., Martin, A. E. Jr., & Conrad, C. M. (1959). An empirical method for estimating the degree of crystallinity of native cellulose using the X-Ray diffractometer. Textile Research Journal, 29(10), 786-794. https://doi.org/10.1177/004051755902901003

Syafri, E., Jamaluddin, Sari, N. H., Mahardika, M., Amanda, P., & Ilyas, R. A. (2022). Isolation and characterization of cellulose nanofibers from Agave gigantea by chemical-mechanical treatment. International Journal of Biological Macromolecules, 200, 25-33. https://doi.org/10.1016/j.ijbiomac.2021.12.111

Tamo, A. K., Doench, I., Deffo, G., Jiokeng, S. L. Z., Doungmo, G., Ghislain Fotsop, C., Temgoua, R. C. T., Montembault, A., Serghei, A., Njanja, E., Tonle, I. K., & Osorio-Madrazo, A. (2025). Lignocellulosic biomass and its main structural polymers as sustainable materials for (bio)sensing applications. Journal of Materials Chemistry A, 13(30), 24185-24253. https://doi.org/10.1039/D5TA02900G

Tippmann, S., Anfelt, J., David, F., Rand, J. M., Siewers, V., Uhlén, M., Nielsen, J., & Hudson, E. P. (2017). Affibody scaffolds improve sesquiterpene production in Saccharomyces cerevisiae. En ACS Synthetic Biology, 6(1), 19-28. https://doi.org/10.1021/acssynbio.6b00109

Valenzuela-Cobos, J. D., Pérez-Martínez, S., Fiallos-Cárdenas, M., & Guevara-Viejó, F. (2024). Data mining for the characterization of a paper prototype obtained with bacterial cellulose derived from banana and pineapple by-products. Applied Sciences, 14(23), 11426. https://doi.org/10.3390/app142311426

Viri, I. (2025, 8 de agosto). Standard methods for the examination of water and wastewater, 24rd edition (2022). Standardlab. https://www.standardlab.com.ec/post/standard-methods-for-the-examination-of-water-and-wastewater-24rd-edition-2022

Wang, S.-S., Han, Y.-H., Ye, Y.-X., Shi, X.-X., Xiang, P., Chen, D.-L., & Li, M. (2017). Physicochemical characterization of high-quality bacterial cellulose produced by Komagataeibacter sp. Strain W1 and identification of the associated genes in bacterial cellulose production. RSC Advances (71), 45145-45155. https://doi.org/10.1039/c7ra08391b

Xu, L., Wang, Y.-Y., Huang, J., Chen, C.-Y., Wang, Z.-X., & Xie, H. (2020). Silver nanoparticles: Synthesis, medical applications and biosafety. Theranostics, 10(20), 8996-9031. https://doi.org/10.7150/thno.45413

Zahan, K. A., Pa’e, N., & Muhamad, I. I. (2015). Monitoring the effect of pH on bacterial cellulose production and Acetobacter xylinum 0416 growth in a rotary discs reactor. Arabian Journal for Science and Engineering, 40(7), 1881-1885. https://doi.org/10.1007/s13369-015-1712-z

Zaini, H. M., Saallah, S., Roslan, J., Sulaiman, N. S., Munsu, E., Wahab, N. A., & Pindi, W. (2023). Banana biomass waste: A prospective nanocellulose source and its potential application in food industry–A review. Heliyon, 9(8), e18734. https://doi.org/10.1016/j.heliyon.2023.e18734

Zou, D., & Fan, Q. (2022). Present situation of globle banana production and trade and prospect for banana industry. Guangdong Agricultural Sciences, 49(7), 131-140. https://doi.org/10.16768/j.issn.1004-874X.2022.07.017

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Published

2026-06-15

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No. 50 (2026)

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Science and technology

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How to Cite

Zeas Sesme, A. J., Martínez Castillo , T. J. ., & Fiallos Cárdenas, M. A. (2026). Production and optimisation of bacterial cellulose from banana (musa spp.) banana rachis juice pre-treated with ozone using a box–behnken design. Ingeniería Industrial, 50, 281-306. https://doi.org/10.26439/ing.ind2026.n50.8611