TINJAUAN BIOKIMIA ENZIM INVERTASE PADA HIDROLISIS SUKROSA: Mekanisme Katalitik, Kinetika, Faktor Fisikokimia, dan Implikasi Aplikatif
Abstract
Invertase (beta-fructofuranosidase; EC 3.2.1.26) is an enzyme that catalyzes the hydrolysis of sucrose into glucose and fructose and plays a crucial role in carbohydrate metabolism, fermentation, the food industry, and biotechnology. This review article synthesizes studies on invertase classification, the sucrose hydrolysis reaction, the catalytic mechanism, the Michaelis-Menten kinetic model, the determination of kinetic parameters such as Km, Vmax, kcat, and activation energy, as well as the influence of physicochemical factors such as pH, temperature, metal ions, inhibitors, and enzyme shape on its activity and stability. In general, invertase belongs to the glycoside hydrolase family 32 and operates through a retaining double-displacement mechanism involving aspartate and glutamate residues in the active site. Changing substrate concentration increases the reaction rate to near Vmax, but real-world systems can deviate due to substrate inhibition, product inhibition, diffusion barriers, and the influence of the reaction matrix. Increasing temperature increases the reaction rate constant up to the optimum temperature, but excessive temperatures cause denaturation and deviation from the Arrhenius model. Most invertases exhibit an optimum pH in the weak acid range, although alkaline invertases and low-temperature active invertases with different characteristics exist. Various analytical methods, such as DNS, Nelson-Somogyi, Bradford, chromatography, SDS-PAGE, as well as spectroscopic and crystallographic approaches, have been used to assess the activity, purity, and molecular properties of invertases. Overall, this review demonstrates that invertase performance is a result of the interaction between protein structure, reaction conditions, and the chemical environment. Therefore, an integrated understanding of these aspects is essential for optimizing laboratory and industrial applications(Manoochehri et al., 2020 ; Lammens et al., 2009 ; Potrich & Amaral, 2018).
Keywords: invertase, sucrose, hydrolysis, enzyme kinetics, physicochemical factors
Downloads
References
Adnadjevic, B. K., & Jovanovic, J. (2012). A comparative kinetics study on the isothermal heterogeneous acid-catalyzed hydrolysis of sucrose under conventional and microwave heating. Journal of Molecular Catalysis A: Chemical, 70-77. https://doi.org/10.1016/j.molcata.2011.12.027
Al Salhen, K. (2022). Biochemical study on the kinetic properties of the invertase produced by Saccharomyces cerevisiae. Scientific Journal for Faculty of Science-Sirte University, 2(2), 1-11. https://doi.org/10.37375/sjfssu.v2i2.75
Al-Odat, I. (2024). Educational activity of enzyme kinetics in an undergraduate biochemistry course: Invertase enzyme as a model. Journal of Microbiology & Biology Education, 25(2), e00050-24. https://doi.org/10.1128/jmbe.00050-24
Avila, T., et al. (2022). Extraction, purification and characterization of invertase from pineapple. Revista Brasileira de Fruticultura, 44(2). https://doi.org/10.1590/0100-29452022849
Awad, G. E. A., Amer, H., El-Gammal, E. W., Helmy, W. A., Esawy, M. A., & Elnashar, M. M. M. (2013). Production optimization of invertase by Lactobacillus brevis Mm-6 and its immobilization on alginate beads. Carbohydrate Polymers, 93(2), 740–746. https://doi.org/10.1016/j.carbpol.2012.12.039
Bassetti, F. J., Bergamasco, R., de Moraes, F. F., & Zanin, G. M. (2000). Thermal stability and deactivation energy of free and immobilized invertase. Brazilian Journal of Chemical Engineering, 17(4-7). https://doi.org/10.1590/S0104-66322000000400050
Bergamasco, R., Bassetti, F. J., de Moraes, F. F., & Zanin, G. M. (2000). Characterization of free and immobilized invertase regarding activity and energy of activation. Brazilian Journal of Chemical Engineering, 17(4-7). https://doi.org/10.1590/S0104-66322000000400051
Chen, Z., Zhang, D., Liu, H., et al. (2021). Identification of an invertase with high specific activity for raffinose hydrolysis and its application in soymilk treatment. Frontiers in Microbiology, 12. https://doi.org/10.3389/fmicb.2021.646801
Cho, Y.-S., & Lim, H.-S. (2018). Comparison of various estimation methods for the parameters of Michaelis-Menten equation based on in vitro elimination kinetic simulation data. Translational and Clinical Pharmacology, 26(1), 39-47. https://doi.org/10.12793/tcp.2018.26.1.39
Choi, B., Rempala, G. A., & Kim, J. K. (2017). Beyond the Michaelis-Menten equation: Accurate and efficient estimation of enzyme kinetic parameters. Scientific Reports, 7(1), 17018. https://doi.org/10.1038/s41598-017-17072-z
Cornish-Bowden, A. (2015). One hundred years of Michaelis-Menten kinetics. Perspectives in Science, 4, 3-9. https://doi.org/10.1016/j.pisc.2014.12.002
Davidia, D., Noorb, E., Liebermeisterc, W., Bar-Evend, A., Flamholze, A., Tummlerf, K., Barenholza, U., Goldenfelda, M., Shlomig, T., & Miloa, R. (2016). Global characterization of in vivo enzyme catalytic rates and their correspondence to in vitro kcat measurements. Proceedings of the National Academy of Sciences of the United States of America, 113(12), 3401–3406. https://doi.org/10.1073/pnas.1514240113
2.png)
