Enhanced Antibacterial and Anticancer Activities of Soy Protein Isolate through Conjugation with Iron Oxide and Zinc Oxide Nanoparticles

Ramy M. Romeilah; Solafa Abdelhakim; Mohamed Raafat Atteya; Sayed E. Younis; Khaled M. A. Ramadan; Hossam S. El-Beltagi; Gamal Enan; Ahmed Maher Gaber; Mahmoud M. El-saber; Nada Ali AboZaid; Ali Osman

Authors

Ramy M. Romeilah Department of Chemistry, College of Science, University of Hail, Saudi Arabia
Solafa Abdelhakim Department of Plant Production (Microbiology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt
Mohamed Raafat Atteya Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia
Sayed E. Younis Department of Plant Production (Plant Pathology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt
Khaled M. A. Ramadan Central Laboratories, Department of Chemistry, King Faisal University, Al-Ahsa 31982, Saudi Arabia
Hossam S. El-Beltagi Agricultural Biotechnology Department, College of Agriculture and Food Sciences, King Faisal University, Al-Ahsa 31982, Saudi Arabia
Gamal Enan Department of Botany and Microbiology, Faculty of Science, Zagazig University, Zagazig, 44519, Egypt
Ahmed Maher Gaber Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia
Mahmoud M. El-saber Biochemistry Unit, Genetic Resources Department, Desert Research Center, El-Matarya, Cairo 11753, Egypt
Nada Ali AboZaid Department of Health Management, College of Public Health and Health Informatics, University of Hail, Hail, Saudi Arabia
Ali Osman Department of Biochemistry, Faculty of Agriculture, Zagazig University, Zagazig, 44519, Egypt

Keywords:

Green synthesis, Nanocomposites, Apoptosis, Caspase activation, Membrane disruption, Biocompatible carrier

Abstract

Soy protein isolate (SPI) was utilized as a biocompatible matrix for the development of Fe₃O₄ and ZnO nanoparticle composites with enhanced biological activities. The synthesized nanocomposites were characterized using various analytical methods that confirmed the successful nanoparticle formation and conjugation with SPI. Antibacterial activity was evaluated against L. monocytogenes, E. coli, and B. cereus using agar well diffusion, minimum inhibitory concentration (MIC), growth kinetics, crystal violet uptake, and cell lysis assays. Both nanocomposites exhibited superior antibacterial efficacy compared with SPI alone, with ZnO-based composites demonstrating the strongest broad-spectrum activity. Nanoformulation reduced MIC values by up to eightfold and significantly enhanced membrane permeability and bacteriolytic effects. The anticancer potential of tested materials was assessed against A549 and HCT116 cell lines. ZnO-NPs-SPI showed the highest cytotoxicity, producing IC₅₀ values of 27.8 and 31.25 µg/mL against A549 and HCT116 cells, respectively. Furthermore, treatment induced significant activation of caspase-3 and caspase-9, indicating apoptosis-mediated cell death. These findings demonstrate that SPI-based nanocomposites, particularly those containing ZnO-NPs, represent promising multifunctional platforms for antimicrobial and anticancer applications.

Author Biographies

Ramy M. Romeilah , Department of Chemistry, College of Science, University of Hail, Saudi Arabia

Department of Chemistry, College of Science, University of Hail, Saudi Arabia

Solafa Abdelhakim , Department of Plant Production (Microbiology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt

Department of Plant Production (Microbiology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt

Mohamed Raafat Atteya , Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia

Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia

Sayed E. Younis , Department of Plant Production (Plant Pathology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt

Department of Plant Production (Plant Pathology Specialization), Faculty of Development and Technology, Zagazig University, Zagazig, 44519, Egypt

Khaled M. A. Ramadan , Central Laboratories, Department of Chemistry, King Faisal University, Al-Ahsa 31982, Saudi Arabia

Central Laboratories, Department of Chemistry, King Faisal University, Al-Ahsa 31982, Saudi Arabia

Gamal Enan , Department of Botany and Microbiology, Faculty of Science, Zagazig University, Zagazig, 44519, Egypt

Department of Botany and Microbiology, Faculty of Science, Zagazig University, Zagazig, 44519, Egypt

Ahmed Maher Gaber , Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia

Department of Physical Therapy, College of Applied Medical Sciences, University of Hail, Hail 81451, Saudi Arabia

Mahmoud M. El-saber , Biochemistry Unit, Genetic Resources Department, Desert Research Center, El-Matarya, Cairo 11753, Egypt

Biochemistry Unit, Genetic Resources Department, Desert Research Center, El-Matarya, Cairo 11753, Egypt

Nada Ali AboZaid , Department of Health Management, College of Public Health and Health Informatics, University of Hail, Hail, Saudi Arabia

Department of Health Management, College of Public Health and Health Informatics, University of Hail, Hail, Saudi Arabia

Ali Osman , Department of Biochemistry, Faculty of Agriculture, Zagazig University, Zagazig, 44519, Egypt

Department of Biochemistry, Faculty of Agriculture, Zagazig University, Zagazig, 44519, Egypt