Aquaculture Sciences

Aquaculture Sciences

Evaluation of the antimicrobial activity of recombinant Goldfish (Carassius auratus) Pro-Hepcidin against gram-positive and gram-negative bacteria

Document Type : Original Article

Authors
Maryam Gheisari 1
Fatemeh Peyravii Ghadikolaii 1
Rahem Khoshbakht 2
Poulin Shohreh 3
Hami Kaboosi 4
1 Department of Biology, QaS.C., Islamic Azad University, Qaemshahr, Iran.
2 Department of Pathobiology, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran.
3 Department of Clinical Sciences, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran.
4 Department of Microbiology, Am.C., Islamic Azad University, Amol, Iran.
Abstract
The increasing prevalence of antibiotic-resistant pathogenic bacteria has highlighted the urgent need for alternative antimicrobial agents. Antimicrobial peptides, including hepcidin, are key components of the innate immune system of vertebrates and represent promising candidates in this context. The present study aimed to produce full-length recombinant pro-hepcidin from goldfish (Carassius auratus) and to evaluate its antibacterial activity against selected Gram-positive and Gram-negative bacteria. For this purpose, the complete pro-hepcidin gene sequence was retrieved, codon-optimized, fused with a histidine tag, cloned into the pET28a expression vector, and expressed in Escherichia coli (BL21) Rosetta. The recombinant protein was subsequently purified and its antimicrobial activity was assessed against Salmonella enterica (Gram-negative), Staphylococcus aureus, and Lactococcus garvieae (Gram-positive) using the disk diffusion method and determination of the minimum inhibitory concentration (MIC). The results demonstrated that recombinant pro-hepcidin exhibited significant inhibitory effects against all tested bacteria, with MIC values of 50, and 25 µg/mL for S. enterica, S. aureus, and L. garvieae, respectively. Moreover, clear and statistically significant inhibition zones were observed in the disk diffusion assay, even against bacterial strains showing resistance to conventional antibiotics. Overall, these findings indicate that recombinant goldfish pro-hepcidin possesses broad-spectrum antibacterial activity and may serve as a promising candidate for the development of novel antimicrobial agents as alternatives to antibiotics, particularly in medical and aquaculture application.
Keywords
Pro-Hepcidin
Gold fish
Aquaculture pathogens
Antibacterial activity
Alternative to antibiotics
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Aquaculture Sciences
Volume 13, Issue 2
October 2025
Pages 91-99