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Feb 17, 2025
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May 14, 2025
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Abstract
The objective of this study was to compare the properties and performance of invertase enzyme isolated from baker's yeast, both in free and immobilized form on a starch-copper nanocomposite (SCN). The SCN was synthesized using starch as a reducing agent for the biological production of copper nanoparticles (CuNPs). The Characterization of SCN was performed using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), and X-ray diffraction to confirm nanoparticle formation and structural properties. The immobilization of invertase onto SCN was optimized by varying nanoparticle concentration, pH, incubation time, and temperature to maximize enzyme attachment and activity. Enzyme activity was measured for both free and immobilized forms to determine the immobilization efficiency. The study found that the high levels of enzyme immobilization were observed at pH = 9, temperature T = 30, and 3% SCN concentration. For both free and immobilized invertase, the ideal reaction temperatures were 35°C and 40°C, with corresponding pH values of 5 and 4.5. Reusability experiments revealed that the immobilized enzyme retained 49% of its activity after ten cycles, demonstrating improved stability and potential for repeated use. The results suggest that enzyme immobilization on SCN occurs through non-covalent interactions, providing a practical and sustainable approach for biocatalytic applications. This research highlights the potential of starch-based nanocomposites for enzyme stabilization, offering a cost-effective and environmentally friendly solution for industrial and biotechnological applications.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
How to Cite
Chouia, M., & Derouiche, S. (2025). Comparative study of immobilized enzyme on nano-composite (SCN) and free enzyme of invertase isolated from baker’s yeast. Journal of Agriculture and Applied Biology, 6(2), 251-263. https://doi.org/10.11594/jaab.06.02.09
References
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Ferreira, M. V., Rossler, A. F., Toralles, R. P., Ruiz, W. A., & Rombaldi, C. V. (2018). Optimized extraction and partial purification of invertase isolated from S. cerevisiae in peach puree. Brazilian Journal of Fruit Growing, 40, e-489. CrossRef
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Maghraby, Y. R., El-Shabasy, R. M., Ibrahim, A. H., & Azzazy, H. M. E.-S. (2023). Enzyme immobilization technologies and industrial applications. ACS Omega, 8, 5184–5196. CrossRef
Malhotra, I., & Basir, S. F. (2020). Application of invertase immobilized on chitosan using glutaraldehyde or tris(hydroxymethyl)phosphine as cross-linking agent to produce bioethanol. Applied Biochemistry and Biotechnology, 191, 838–851. CrossRef
Marquez, L. D. S., Cabral, B. V., Freitas, F. F., Cardoso, V. L., & Ribeiro, E. J. (2008). Optimization of invertase immobilization by adsorption in ionic exchange resin for sucrose hydrolysis. Journal of Molecular Catalysis B: Enzymatic, 51, 86–92. CrossRef
Martis, P., Fonseca, A., Mekhalif, Z., & Delhalle, J. (2010). Optimization of cuprous oxide nanocrystals deposition on multiwalled carbon nanotubes. Journal of Nanoparticle Research, 12, 439–448. CrossRef
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428. CrossRef
Nadeem, H., Rashid, M. H., Siddique, M. H., Azeem, F., Muzammil, S., Javed, M. R., & Riaz, M. (2015). Microbial invertases: A review on kinetics, thermodynamics, physiochemical properties. Process Biochemistry, 50(8), 1202–1210. CrossRef
Naqash, F., Masoodi, F. A., & Rather, S. A. (2019). Food enzymes and nanotechnology. In M. Kuddus (Ed.), Enzymes in food biotechnology (pp. 769–784). Academic Press. CrossRef
Ouidad, A., Sara, C., & Samir, D. (2020). Biological properties and acute toxicity study of copper oxide nanoparticles prepared by aqueous leaves extract of Portulaca oleracea (L). Asian Journal of Pharmaceutical Research, 10, 89–94. CrossRef
Parapouli, M., Vasileiadis, A., Afendra, A. S., & Hatziloukas, E. (2020). Saccharomyces cerevisiae and its industrial applications. AIMS Microbiology, 6(1), 1–31. CrossRef
Pedrol, N., & Tamayo, P. (2001). Protein content quantification by Bradford method. In J. M. S. Cabral, M. Kalil, & A. L. Carvalho (Eds.), Immobilization of Enzymes and Cells (pp. 283–295). Springer. CrossRef
Razzaghi, M., Homaei, A., Vianello, F., Azad, T., Sharma, T., Nadda, A. K., Stevanato, R., Bilal, M., & Iqbal, H. M. N. (2022). Industrial applications of immobilized nano-biocatalysts. Bioprocess and Biosystems Engineering, 45, 237–256. CrossRef
Remy, E., Niño-González, M., Godinho, C. P., Cabrito, T. R., Matos, R. G., dos Santos, T. C., Sánchez-Fernández, R., & Sa-Correia, I. (2017). Heterologous expression of the yeast Tpo1p or Pdr5p membrane transporters in Arabidopsis confers plant xenobiotic tolerance. Scientific Reports, 7, 4529. CrossRef
Rouzer, C. A., & Marnett, L. J. (2020). Structural and chemical biology of the interaction of cyclooxygenase with substrates and non-steroidal anti-inflammatory drugs. Chemical Reviews, 120, 7592–7641. CrossRef
Sachin, H. R., Hosamani, P., & Ganeshprasad, D. N., & Sneharani, A. H. (2020). Immobilization of trypsin enzyme on silver nanoparticles. Biomedicine, 40(2), 188–191. Direct Link.
Samir, D., Serin, B., Manal, B., Lina, B., & Djoumana, A. (2022). Eco-friendly phytosynthesis of copper nanoparticles using Medicago sativa extract: A biological activity and acute toxicity evaluation. World Journal of Environmental Biosciences, 11, 9–15. CrossRef
Sjölin, M., Djärf, M., Ismail, M., Schagerlöf, H., Wallberg, O., Hatti-Kaul, R., & Sayed, M. (2024). Investigating the Inhibitory Factors of Sucrose Hydrolysis in Sugar Beet Molasses with Yeast and Invertase. Catalysts, 14(5), 330. CrossRef
Wang, Z., Li, H., & Weng, Y. (2024). A neutral invertase controls cell division besides hydrolysis of sucrose for nutrition during germination and seed setting in rice. iScience, 27(7), 110217. CrossRef
Waseda, Y., Matsubara, E., & Shinoda, K. (2011). X-ray diffraction crystallography: Introduction, examples and solved problems. Springer Science & Business Media. CrossRef
Amaya-Delgado, L., Hidalgo-Lara, M. E., & Montes-Horcasitas, M. C. (2006). Hydrolysis of sucrose by invertase immobilized on nylon-6 microbeads. Food Chemistry, 99, 299–304. CrossRef
Aram Akram Mohammed, Tariq Abubakr Ahmad, Ibrahim Maaroof Noori, Rasul Rafiq Aziz, & Karim Ahmad. (2020). Application of baking yeast to induce rooting in hardwood cuttings of olive (Olea europaea L.) cv. Sorani. Euphrates Journal of Agriculture Science, 12(2), 274–280. Direct Link.
Bayda, S., Adeel, M., Tuccinardi, T., Cordani, M., & Rizzolio, F. (2019). The history of nanoscience and nanotechnology: From chemical-physical applications to nanomedicine. Molecules, 25(1), 112. CrossRef
Fenangad, D., & Orge, R. F. (2015). Simple seed coating technology for improved seedling establishment in direct-seeded rice. OIDA International Journal of Sustainable Development, 8(11), 35-42. Direct Link.
Fernandez-San Millan, A., Farran, I., Larraya, L., Ancin, M., Arregui, L. M., & Veramendi, J. (2020). Plant growth-promoting traits of yeasts isolated from Spanish vineyards: Benefits for seedling development. Microbiological Research, 237, 126480. CrossRef
Ferreira, M. V., Rossler, A. F., Toralles, R. P., Ruiz, W. A., & Rombaldi, C. V. (2018). Optimized extraction and partial purification of invertase isolated from S. cerevisiae in peach puree. Brazilian Journal of Fruit Growing, 40, e-489. CrossRef
Gawande, M. B., Goswami, A., Felpin, F. X., Asefa, T., Huang, X., Silva, R., Zou, X., Zboril, R., & Varma, R. S. (2016). Cu and Cu-based nanoparticles: Synthesis and applications in catalysis. Chemical Reviews, 116, 3722–3811. CrossRef
Hakkoymaz, O., & Mazi, H. (2020). An immobilized invertase enzyme for the selective determination of sucrose in fruit juices. Analytical Biochemistry, 611, 27. CrossRef
Hartemann, P., Hoet, P., Proykova, A., Fernandes, T., Baun, A., De Jong, W., Filser, J., Hensten, A., Kneuer, C., Maillard, J. Y., Norppa, H., Scheringer, M., & Wijnhoven, S. (2015). Nanosilver: Safety, health and environmental effects and role in antimicrobial resistance. Materials Today, 18, 122–123. CrossRef
Khan, A., Rashid, A., Younas, R., & Chong, R. (2016). A chemical reduction approach to the synthesis of copper nanoparticles. International Nano Letters, 6, 21–26. CrossRef
Kulp, K. (1975). Carbohydrases. In G. Reed (Ed.), Enzymes in food processing (2nd ed., pp. 53–122). Academic Press. CrossRef
Kumar, A., Asthana, M., Jain, K. G., & Singh, V. (2017). Microbial production of enzymes: An overview. In G. Vijai Kumar, S. Zuhlke, E. X. F. Filho, M. C. D. Duarte-de-Brito, & D. P. (Eds.), Microbial applications (pp. 107–138). De Gruyter. CrossRef
Kumar, V. V. (2016). Plant growth-promoting microorganisms: Interaction with plants and soil. In K. Hakeem, M. Akhtar, & S. Abdullah (Eds.), Plant, Soil and Microbes (pp. 1-20). Spring-er. CrossRef
Maghraby, Y. R., El-Shabasy, R. M., Ibrahim, A. H., & Azzazy, H. M. E.-S. (2023). Enzyme immobilization technologies and industrial applications. ACS Omega, 8, 5184–5196. CrossRef
Malhotra, I., & Basir, S. F. (2020). Application of invertase immobilized on chitosan using glutaraldehyde or tris(hydroxymethyl)phosphine as cross-linking agent to produce bioethanol. Applied Biochemistry and Biotechnology, 191, 838–851. CrossRef
Marquez, L. D. S., Cabral, B. V., Freitas, F. F., Cardoso, V. L., & Ribeiro, E. J. (2008). Optimization of invertase immobilization by adsorption in ionic exchange resin for sucrose hydrolysis. Journal of Molecular Catalysis B: Enzymatic, 51, 86–92. CrossRef
Martis, P., Fonseca, A., Mekhalif, Z., & Delhalle, J. (2010). Optimization of cuprous oxide nanocrystals deposition on multiwalled carbon nanotubes. Journal of Nanoparticle Research, 12, 439–448. CrossRef
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31, 426–428. CrossRef
Nadeem, H., Rashid, M. H., Siddique, M. H., Azeem, F., Muzammil, S., Javed, M. R., & Riaz, M. (2015). Microbial invertases: A review on kinetics, thermodynamics, physiochemical properties. Process Biochemistry, 50(8), 1202–1210. CrossRef
Naqash, F., Masoodi, F. A., & Rather, S. A. (2019). Food enzymes and nanotechnology. In M. Kuddus (Ed.), Enzymes in food biotechnology (pp. 769–784). Academic Press. CrossRef
Ouidad, A., Sara, C., & Samir, D. (2020). Biological properties and acute toxicity study of copper oxide nanoparticles prepared by aqueous leaves extract of Portulaca oleracea (L). Asian Journal of Pharmaceutical Research, 10, 89–94. CrossRef
Parapouli, M., Vasileiadis, A., Afendra, A. S., & Hatziloukas, E. (2020). Saccharomyces cerevisiae and its industrial applications. AIMS Microbiology, 6(1), 1–31. CrossRef
Pedrol, N., & Tamayo, P. (2001). Protein content quantification by Bradford method. In J. M. S. Cabral, M. Kalil, & A. L. Carvalho (Eds.), Immobilization of Enzymes and Cells (pp. 283–295). Springer. CrossRef
Razzaghi, M., Homaei, A., Vianello, F., Azad, T., Sharma, T., Nadda, A. K., Stevanato, R., Bilal, M., & Iqbal, H. M. N. (2022). Industrial applications of immobilized nano-biocatalysts. Bioprocess and Biosystems Engineering, 45, 237–256. CrossRef
Remy, E., Niño-González, M., Godinho, C. P., Cabrito, T. R., Matos, R. G., dos Santos, T. C., Sánchez-Fernández, R., & Sa-Correia, I. (2017). Heterologous expression of the yeast Tpo1p or Pdr5p membrane transporters in Arabidopsis confers plant xenobiotic tolerance. Scientific Reports, 7, 4529. CrossRef
Rouzer, C. A., & Marnett, L. J. (2020). Structural and chemical biology of the interaction of cyclooxygenase with substrates and non-steroidal anti-inflammatory drugs. Chemical Reviews, 120, 7592–7641. CrossRef
Sachin, H. R., Hosamani, P., & Ganeshprasad, D. N., & Sneharani, A. H. (2020). Immobilization of trypsin enzyme on silver nanoparticles. Biomedicine, 40(2), 188–191. Direct Link.
Samir, D., Serin, B., Manal, B., Lina, B., & Djoumana, A. (2022). Eco-friendly phytosynthesis of copper nanoparticles using Medicago sativa extract: A biological activity and acute toxicity evaluation. World Journal of Environmental Biosciences, 11, 9–15. CrossRef
Sjölin, M., Djärf, M., Ismail, M., Schagerlöf, H., Wallberg, O., Hatti-Kaul, R., & Sayed, M. (2024). Investigating the Inhibitory Factors of Sucrose Hydrolysis in Sugar Beet Molasses with Yeast and Invertase. Catalysts, 14(5), 330. CrossRef
Wang, Z., Li, H., & Weng, Y. (2024). A neutral invertase controls cell division besides hydrolysis of sucrose for nutrition during germination and seed setting in rice. iScience, 27(7), 110217. CrossRef
Waseda, Y., Matsubara, E., & Shinoda, K. (2011). X-ray diffraction crystallography: Introduction, examples and solved problems. Springer Science & Business Media. CrossRef