Immobilization of Kluyveromyces lactis for xylose production using different lignocellulosic materials as carbon sources

Authors

  • Dharmitha Chandran Faculty of Industrial Sciences and Technology, University Malaysia Pahang Al-Sultan Abdullah, Lebuh Persiaran Tun Khalil Yaakob, 26300 Kuantan, Pahang, Malaysia , Universiti Malaysia Pahang Al-Sultan Abdullah image/svg+xml https://orcid.org/0009-0007-5551-0728
  • Mohd Azwan Jenol Faculty of Industrial Sciences and Technology, University Malaysia Pahang Al-Sultan Abdullah, Lebuh Persiaran Tun Khalil Yaakob, 26300 Kuantan, Pahang, Malaysia , Universiti Malaysia Pahang Al-Sultan Abdullah image/svg+xml
  • Shoriya Aruni Abdul Manaf Faculty of Industrial Sciences and Technology, University Malaysia Pahang Al-Sultan Abdullah, Lebuh Persiaran Tun Khalil Yaakob, 26300 Kuantan, Pahang, Malaysia , Universiti Malaysia Pahang Al-Sultan Abdullah image/svg+xml https://orcid.org/0000-0001-7255-6890

DOI:

https://doi.org/10.15282/cst.v6i1.13288

Keywords:

Kluyveromyces lactis, Cell immobilization, Lignocellulosic biowaste

Abstract

Lignocellulosic biowaste is a renewable and abundant resource with significant potential for the production of value-added products such as xylose which is a major intermediary in the biofuel, bioplastic, and pharmaceutical sectors. However, problems such as inefficient bioconversion and instability of free-cell fermentation systems limit its industrial application. This research investigates the use of immobilized recombinant Kluyveromyces lactis for xylose production using various lignocellulosic biowastes such as sugarcane bagasse, corn stover, rice straw, oil palm fronds, and banana leaves. The biowastes were washed, dried, ground, and pre-treated with 1% sodium hydroxide (NaOH) solution. K. lactis cells were cultivated in yeast peptone dextrose (YPD) medium and immobilized in 2% sodium alginate beads cross-linked with 0.1 M calcium chloride (CaCl2). Fermentation was carried out at 30 °C and 150 rpm for 24 hours using 1 g of pre-treated biomass and immobilized yeast beads in 100 mL of YPD medium. Reducing sugar concentration was determined using the dinitro salicylic acid (DNS) method, while yeast growth and pH changes were monitored. Results demonstrated that immobilized K. lactis could produce xylose production across all biowaste types, with sugarcane bagasse and corn stover yielding the highest reducing sugar concentrations. However, a further optimization is needed to increase the enzyme production from the immobilized cell systems. The findings highlight a cost-effective and environmentally friendly approach for improving industrial xylose production through K. lactis immobilization on lignocellulosic biowastes.

References

[1] M. Mujtaba, L. F. Fraceto, M. Fazeli, et al., “Lignocellulosic biomass from agricultural waste to the circular economy: A review with focus on biofuels, biocomposites and bioplastics,” Journal of Cleaner Production, vol. 402, p. 136815, 2023, https://doi.org/10.1016/j.jclepro.2023.136815.

[2] E. Palmqvist and B. Hahn-Hägerdal, “Fermentation of lignocellulosic hydrolysates. I: Inhibition and detoxification,” Bioresource Technology, vol. 74, no. 1, pp. 17–24, 2000, https://doi.org/10.1016/S0960-8524(99)00160-1.

[3] B. B. Ying, J. Cai, X. Gao, et al., “Isolation, identification, and tolerance analysis of yeast during the natural fermentation process of Sidamo coffee beans,” Archives of Microbiology, vol. 206, no. 6, p. 279, 2024, https://doi.org/10.1007/s00203-024-04017-0.

[4] J. Ruchala and A. A. Sibirny, “Pentose metabolism and conversion to biofuels and high-value chemicals in yeasts,” FEMS Microbiology Reviews, vol. 45, no. 4, pp. 1–44, 2021, https://doi.org/10.1093/femsre/fuaa069.

[5] M. Baptista and L. Domingues, “Kluyveromyces marxianus as a microbial cell factory for lignocellulosic biomass valorization,” Biotechnology Advances, vol. 60, p. 108027, 2022, https://doi.org/10.1016/j.biotechadv.2022.108027.

[6] W. Tischer and F. Wedekind, “Immobilized enzymes: Methods and applications,” in Biocatalysis — From Discovery to Application (Topics in Current Chemistry, vol. 200), Berlin, Germany: Springer, 1999, pp. 95–126, https://doi.org/10.1007/3-540-68116-7_4.

[7] M. Becerra, B. Baroli, A. M. Fadda, J. B. Méndez, and M. G. Siso, “Lactose bioconversion by calcium-alginate immobilization of Kluyveromyces lactis cells,” Enzyme and Microbial Technology, vol. 29, no. 8–9, pp. 506–512, 2001, https://doi.org/10.1016/S0141-0229(01)00409-4.

[8] M. N. Kumar, A. I. Gialleli, J. B. Masson, P. Kandylis, A. Bekatorou, A. A. Koutinas, and M. Kanellaki, “Lactic acid fermentation by cells immobilised on various porous cellulosic materials and their alginate/poly-lactic acid composites,” Bioresource Technology, vol. 165, pp. 332–335, 2014, https://doi.org/10.1016/j.biortech.2014.02.110.

[9] J. Zou and X. Chang, “Past, present, and future perspectives on whey as a promising feedstock for bioethanol production by yeast,” Journal of Fungi, vol. 8, no. 4, p. 395, 2022, https://doi.org/10.3390/jof8040395.

[10] S. Vijayaram, R. Sinha, C. Faggio, E. Ringø, and C. C. Chou, “Biopolymer encapsulation for improved probiotic delivery: Advancements and challenges,” AIMS Microbiology, vol. 10, no. 4, p. 986, 2024, https://doi.org/10.3934/microbiol.2024043.

[11] Y. C. Park and J. S. Kim, “Comparison of various alkaline pretreatment methods of lignocellulosic biomass,” Energy, vol. 47, no. 1, pp. 31–35, 2012, https://doi.org/10.1016/j.energy.2012.08.010.

[12] S. F. Z. M. Fuzi, N. M. Mahadi, J. M. Jahim, et al., “Development and validation of a medium for recombinant endo-β-1,4-xylanase production by Kluyveromyces lactis using a statistical experimental design,” Annals of Microbiology, vol. 62, no. 1, pp. 283–292, 2012, https://doi.org/10.1007/s13213-011-0258-x.

[13] I. S. Snoek and H. Y. Steensma, “Why does Kluyveromyces lactis not grow under anaerobic conditions? Comparison of essential anaerobic genes of Saccharomyces cerevisiae with the Kluyveromyces lactis genome,” FEMS Yeast Research, vol. 6, no. 3, pp. 393–403, 2006, https://doi.org/10.1111/j.1567-1364.2005.00007.x.

[14] D. Portugal-Nunes, V. Sànchez i Nogué, S. R. Pereira, S. C. Craveiro, A. J. Calado, and A. M. Xavier, “Effect of cell immobilization and pH on Scheffersomyces stipitis growth and fermentation capacity in rich and inhibitory media,” Bioresources and Bioprocessing, vol. 2, no. 1, p. 13, 2015, https://doi.org/10.1186/s40643-015-0042-z.

[15] M. Heitger and F. Baltar, “Respiration, production, and growth efficiency of marine pelagic fungal isolates,” Journal of Fungi, vol. 9, no. 4, p. 417, 2023, https://doi.org/10.3390/jof9040417.

[16] I. P. Wood, A. Elliston, P. Ryden, I. Bancroft, I. N. Roberts, and K. W. Waldron, “Rapid quantification of reducing sugars in biomass hydrolysates: Improving the speed and precision of the dinitrosalicylic acid assay,” Biomass and Bioenergy, vol. 44, pp. 117–121, 2012, https://doi.org/10.1016/j.biombioe.2012.05.003.

[17] X. Yang, M. Tu, R. Xie, S. Adhikari, and Z. Tong, “A comparison of three pH control methods for revealing effects of undissociated butyric acid on specific butanol production rate in batch fermentation of Clostridium acetobutylicum,” AMB Express, vol. 3, p. 3, 2013, https://doi.org/10.1186/2191-0855-3-3.

[18] S. W. Park, B. H. Kang, H. M. Lee, et al., “Efficient brazzein production in yeast (Kluyveromyces lactis) using a chemically defined medium,” Bioprocess and Biosystems Engineering, vol. 44, no. 4, pp. 913–925, 2021, https://doi.org/10.1007/s00449-020-02499-y.

[19] N. Fukuda, “Apparent diameter and cell density of yeast strains with different ploidy,” Scientific Reports, vol. 13, no. 1, Art. no. 1513, 2023, https://doi.org/10.1038/s41598-023-28800-z.

[20] W. T. Godbey, “Cell growth,” in Biotechnology and Its Applications, 2nd ed. London, U.K.: Academic Press, 2022, pp. 117–150.

[21] P. F. Stanbury, A. Whitaker, and S. J. Hall, Principles of Fermentation Technology, 3rd ed. Oxford, U.K.: Butterworth-Heinemann, 2017, https://doi.org/10.1016/C2013-0-00186-7.

[22] M. Werner-Washburne, E. Braun, G. C. Johnston, and R. A. Singer, “Stationary phase in the yeast Saccharomyces cerevisiae,” Microbiological Reviews, vol. 57, no. 2, pp. 383–401, 1993, https://doi.org/10.1128/mr.57.2.383-401.1993.

[23] C. Duc, M. Pradal, I. Sanchez, J. Noble, C. Tesniere, and B. Blondin, “A set of nutrient limitations trigger yeast cell death in a nitrogen-dependent manner during wine alcoholic fermentation,” PLOS ONE, vol. 12, no. 9, p. e0184838, 2017, https://doi.org/10.1371/journal.pone.0184838.

[24] R. Razmovski and V. Vučurović, “Bioethanol production from sugar beet molasses and thick juice using Saccharomyces cerevisiae immobilized on maize stem ground tissue,” Fuel, vol. 92, no. 1, pp. 1–8, 2012, https://doi.org/10.1016/j.fuel.2011.07.046.

[25] M. F. S. Khan, M. Akbar, Z. Xu, and H. Wang, “A review on the role of pretreatment technologies in the hydrolysis of lignocellulosic biomass of corn stover,” Biomass and Bioenergy, vol. 155, p. 106276, 2021, https://doi.org/10.1016/j.biombioe.2021.106276.

[26] P. H. Pilkington, A. Margaritis, and N. A. Mensour, “Mass transfer characteristics of immobilized cells used in fermentation processes,” Critical Reviews in Biotechnology, vol. 18, no. 2–3, pp. 237–255, 1998, https://doi.org/10.1080/0738-859891224239.

[27] A. E. L. Hesham, Y. S. Mostafa, and L. E. O. AlSharqi, “Optimization of citric acid production by immobilized cells of novel yeast isolates,” Mycobiology, vol. 48, no. 2, pp. 122–132, 2020, https://doi.org/10.1080/12298093.2020.1726854.

[28] Z. Genisheva, J. A. Teixeira, and J. M. Oliveira, “Immobilized cell systems for batch and continuous winemaking,” Trends in Food Science & Technology, vol. 40, no. 1, pp. 33–47, 2014, https://doi.org/10.1016/j.tifs.2014.07.009.

[29] S. A. Abdul Manaf, S. F. Z. Mohamad Fuzi, K. O. Low, et al., “Carbon nanomaterial properties help to enhance xylanase production from recombinant Kluyveromyces lactis through a cell immobilization method,” Applied Microbiology and Biotechnology, vol. 105, no. 21, pp. 8531–8544, 2021, https://doi.org/10.1007/s00253-021-11616-0.

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Published

2026-09-01

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Section

Research Articles

How to Cite

[1]
D. Chandran, M. A. Jenol, and S. A. Abdul Manaf, “Immobilization of Kluyveromyces lactis for xylose production using different lignocellulosic materials as carbon sources”, Curr. Sci. Technol., vol. 6, no. 1, pp. 29–35, Sep. 2026, doi: 10.15282/cst.v6i1.13288.