Evaluation of antibacterial properties of nisin peptide expressed in carrots

• Published in: Scientific reports 2023-12, Vol.13 (1), p.22123-22123, Article 22123
• Main Authors: Fallah Ziarani, Masoumeh, Tohidfar, Masoud, Mirjalili, Mohammad Hossein
• Format: Article
• Language: English
• Subjects: 631/326; 631/449; 631/61; Anti-Bacterial Agents - metabolism; Anti-Bacterial Agents - pharmacology; Bacteria; Daucus carota - genetics; E coli; Enzyme-linked immunosorbent assay; Escherichia coli; Escherichia coli - metabolism; Fruit juices; Fruits; Gene expression; Genomes; Gram-Negative Bacteria; Gram-Positive Bacteria; Humanities and Social Sciences; Lactococcus lactis - metabolism; Liquid chromatography; multidisciplinary; Natural & organic foods; Neomycin; Neomycin phosphotransferase; Nisin; Nucleotide sequence; Peptides; Phosphotransferase; Phosphotransferase II; Plant extracts; Plants, Genetically Modified - genetics; Polymerase chain reaction; Preservatives; Science; Science (multidisciplinary); Staphylococcus aureus; Transgenes; Transgenic plants
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-49466-7

Nisin, derived from Lactococcus lactis , is a well-known natural food preservative. In the present study, the gene of nisin was transformed to carrot by Agrobacterium tumefaciens strain LBA4404 harboring the recombinant binary vector pBI121 containing neomycin phosphotransferase II ( npt II) gene, peptide signal KDEL, and Kozak sequence. The integration of nisin and npt II transgenes into the plant genome was confirmed by polymerase chain reaction (PCR) and dot blot analysis. The gene expression was also performed by RT-PCR and Enzyme-Linked Immunosorbent Assay. The level of nisin expressed in one gram of transgenic plant ranged from 0.05 to 0.08 μg/ml. The stability of nisin varied in orange and peach juices depending on the temperature on the 70th day. The leaf protein extracted from the transgenic plant showed a significant preservative effect of nisin in peach and orange juice. A complete inhibition activity against Staphylococcus aureus and Escherichia coli in orange juice was observed within 24 h. After 24 h, log 1 and log 2 were obtained in a peach juice containing Staphylococcus aureus and Escherichia coli , respectively. Results of HPLC indicated that Chlorogenic and Chicoric acid compounds were increased in transgenic plants, but this increase was not significant. The study of determining the genetic stability of transgenic plants in comparison with non-transgenic plants showed high genetic stability between non-transgenic plants and transgenic plants. This study confirmed the significant inhibitory effect of nisin protein on gram-positive and gram-negative bacteria.