Enhancing Sweetness of Natural Juices Using Partially Purified Invertase from Prune Plum Fruit

Alsoufi, Mohammed

doi:Under indexing

Proceedings of International Conference on Applied Innovation in IT · 2026/06/12 · Vol. 14 · Issue 4 · pp. 819–828

Enhancing Sweetness of Natural Juices Using Partially Purified Invertase from Prune Plum Fruit

Mohammed Abdulrazzaq Alsoufi

📄 Download PDF DOI: Under indexing

Abstract

This study extracted, purified, and immobilized invertase from prune plum (Prunus domestica L.) fruit to increase sweetness by hydrolysis of sucrose and improve the taste of natural orange juice. Invertase (INV) purification process, including the use of ammonium sulfate precipitation at 40-80%, followed by DEAE-cellulose and Sephacryl S-200 chromatography. At the end of purification, the specific activity of INV was 12.84 U/mg, with a 6.83-fold purification and a final yield of 20.41%. INV was immobilized on sodium alginate beads with 86% efficiency. The immobilized invertase (Im-INV) worked best and was most stable at pH 5 and 45°C, and it was stable between 25 and 50°C. It kept full activity for 14 d at 4°C, then dropped to 35% after 30 d. The enzyme also showed strong operational stability, keeping full activity after 17 reuses and 81% activity after 30 reuses. The optimal conditions to increase the sweetness in natural juice are to incubate 100 mL of orange juice with 1 g of Im-INV at 45°C for 30 min.

Keywords

Immobilized Enzyme Invertase Sensory Evaluation Sodium Alginate Sucrose Hydrolysis

References

M. Chalella Mazzocato and J.-C. Jacquier, “Recent advances and perspectives on food-grade immobilisation systems for enzymes,” Foods, vol. 13, no. 13, Art. no. 2127, Jul. 2024, [Online]. Available: https://doi.org/10.3390/foods13132127.
G. Sanjay and S. Sugunan, “Enhanced pH and thermal stabilities of invertase immobilized on montmorillonite K-10,” Food Chem., vol. 94, no. 4, pp. 573-579, Mar. 2006, [Online]. Available: https://doi.org/10.1016/j.foodchem.2004.12.043.
E. Dokuzparmak, “Industrial approach to invertase production from fruit waste for enhanced efficiency and conservation,” ACS Omega, vol. 9, no. 24, pp. 26183-26194, Jun. 2024, [Online]. Available: https://doi.org/10.1021/acsomega.4c01732.
C. C. Liu, L. C. Huang, C. T. Chang, and H. Y. Sung, “Purification and characterization of soluble invertase from suspension-cultured bamboo (Bambusa edulis) cells,” Food Chem., vol. 96, no. 4, pp. 621-631, Jun. 2006, [Online]. Available: https://doi.org/10.1016/j.foodchem.2005.02.044.
F. Amin, H. N. Bhatti, and M. Asgher, “Partial purification and characterization of an acid invertase from Saccharum officinarum L.,” Pak. J. Bot., vol. 42, no. 4, pp. 2531-2540, Aug. 2010, [Online]. Available: https://www.researchgate.net/publication/260096759_Partial_purification_and_characterization_of_an_acid_invertase_from_Saccharum_officinarum_L.
G. Bayramoglu, S. Akgol, A. Bulut, H. Yavuz, and A. Denizli, “Improvement stability and performance of invertase via immobilization onto silanized and polymer brush grafted magnetic nanoparticles,” Food Chem., vol. 221, pp. 1442-1450, Apr. 2017, [Online]. Available: https://doi.org/10.1016/j.foodchem.2016.11.007.
L. M. O. Arruda and M. Vitolo, “Characterization of invertase entrapped into calcium alginate beads,” Appl. Biochem. Biotechnol., vol. 81, no. 1, pp. 23-33, Jul. 1999, [Online]. Available: https://doi.org/10.1385/ABAB:81:1:23.
T. Long, C. C. Milbourn, A. Smith, K. L. S. Khin, A. J. Page, I. Idris, Q. Yang, R. L. Young, and S. Eldeghaidy, “Neural mechanisms and alterations of sweet sensing: Insights from functional magnetic resonance imaging studies,” Life, vol. 15, no. 7, Art. no. 1075, Jul. 2025, [Online]. Available: https://doi.org/10.3390/life15071075.
M. Peng, R. Ginieis, S. Abeywickrema, J. McCormack, and J. Prescott, “Rejection thresholds for sweetness reduction in a model drink predict dietary sugar intake,” Food Qual. Prefer., vol. 110, Art. no. 104965, Aug. 2023, [Online]. Available: https://doi.org/10.1016/j.foodqual.2023.104965.
M. M. Bradford, “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding,” Anal. Biochem., vol. 72, no. 1-2, pp. 248-254, May 1976, [Online]. Available: https://doi.org/10.1016/0003-2697(76)90527-3.
M. J. Alam, M. H. Rahman, and S. M. Abu Sayem, “Extraction, partial purification and characterization of tomato invertase,” Proc. Pakistan Acad. Sci., vol. 44, no. 2, pp. 97-103, Jan. 2007, [Online]. Available: https://www.researchgate.net/publication/284503635_Extraction_partial_purification_and_characterization_of_tomato_invertase.
M. A. Al-Soufi, “Using of immobilized naringinase and pectinase to improvement properties of the natural juice,” Diyala J. Agric. Sci., vol. 8, no. 1, pp. 259-270, Jun. 2016, [Online]. Available: https://journal.djas.uodiyala.edu.iq/index.php/dasj/article/view/1429.
K. Uzun, E. Cevik, and M. Senel, “Invertase immobilization on a metal chelated triazole-functionalized Eupergit® C,” Am. J. Chem., vol. 1, no. 1, pp. 16-21, Oct. 2011, [Online]. Available: https://doi.org/10.5923/j.chemistry.20110101.03.
S. Durakli Velioglu, U. Bağcı, K. G. Güner, H. A. Güleç, and H. M. Velioğlu, “Green purification of invertase from ultrasonicated sifted baker’s yeast by membrane filtration: A comparative study,” Molecules, vol. 30, no. 12, Art. no. 2663, Jun. 2025, [Online]. Available: https://doi.org/10.3390/molecules30122663.
T. L. Avila, R. P. Toralles, E. T. Jansen, M. V. Ferreira, C. R. Kuhn, and W. A. Ruiz, “Extraction, purification and characterization of invertase from Candida guilliermondii isolated from peach solid wastes,” Rev. Bras. Frutic., vol. 44, no. 2, Art. no. e-849, Mar. 2022, [Online]. Available: https://doi.org/10.1590/0100-29452022849.
K. Çetin, I. Perçin, F. Denizli, and A. Denizli, “Tentacle-type immobilized metal affinity cryogel for invertase purification from Saccharomyces cerevisiae,” Artif. Cells Nanomed. Biotechnol., vol. 45, no. 7, pp. 1431-1439, 2017, [Online]. Available: https://doi.org/10.1080/21691401.2016.1243549.
G. Zhou, C. Peng, X. Liu, F. Chang, Y. Xiao, J. Liu, and Z. Fang, “Identification and immobilization of an invertase with high specific activity and sucrose tolerance ability of Gongronella sp. W5 for high fructose syrup preparation,” Front. Microbiol., vol. 11, Art. no. 633, Apr. 2020, [Online]. Available: https://doi.org/10.3389/fmicb.2020.00633.
A. Tanriseven and E. Doğan, “Immobilization of invertase within calcium alginate gel capsules,” Process Biochem., vol. 36, no. 11, pp. 1081-1083, May 2001, [Online]. Available: https://doi.org/10.1016/S0032-9592(01)00146-7.
D. Polanía Melo, A. Hernández Bravo, J. C. Cruz, and L. H. Reyes, “Invertase immobilization on magnetite nanoparticles for efficient fructooligosaccharide generation: A comprehensive kinetic analysis and reactor design strategy,” ChemEngineering, vol. 7, no. 3, Art. no. 55, Jun. 2023, [Online]. Available: https://doi.org/10.3390/chemengineering7030055.
P. P. Waifalkar, S. B. Parit, A. D. Chougale, S. K. Mahajan, and V. A. Bapat, “Immobilization of invertase on chitosan coated γ-Fe₂O₃ magnetic nanoparticles to facilitate magnetic separation,” J. Colloid Interface Sci., vol. 482, pp. 159-164, Nov. 2016, [Online]. Available: https://doi.org/10.1016/j.jcis.2016.07.082.
O. Hakkoymaz and H. Mazı, “Termostable and effective immobilized invertase for sucrose determination in fruit juices,” Anal. Biochem., vol. 690, Art. no. 115515, Jul. 2024, [Online]. Available: https://doi.org/10.1016/j.ab.2024.115515.