Successful use of Nicotiana tabacum hairy roots for the recombinant production of Cecropin A peptide

Biotechnol Appl Biochem. 2022 Jun;69(3):876-886. doi: 10.1002/bab.2158. Epub 2021 May 13.

Abstract

Cecropin A, as an antimicrobial peptide (AMP), is possible to use in medical and agricultural fields as a new and safe biocontrol agent. Therefore, it is highly necessary to find a cost-effective and scalable approach to generate a large scale of it. In this research, the Agrobacterium rhizogenes strain ATCC 15834 was used to transfer the Cecropin A gene to the Nicotiana tabacum. After confirmation of transgenic hairy roots, the antibacterial activity of purified Cecropin A peptide was measured using the agar gel diffusion method. Successful transforming of Cecropin A was confirmed at the RNA and protein levels in hairy root cells using RT-PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The highest Cecropin A amount was detected in line 4 of the transgenic lines using ELISA in comparison with the nontransgenic line. Subsequently, the antimicrobial activity of Cecropin A extracted from line 4 showed the highest inhibition activity against Aspergillus niger. Besides, this activity was stable against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans pathogens after 7 days. The recombinant production of Cecropin A AMP had a yield of 63.81 μg/g of fresh weight. According to a significant yield, this system can be used to produce the Cecropin A peptide for pharmacological and food science applications.

Keywords: Cecropin A; antimicrobial activity; hairy roots; recombinant peptide; stability.

MeSH terms

  • Antimicrobial Cationic Peptides* / pharmacology
  • Escherichia coli / metabolism
  • Nicotiana* / genetics
  • Nicotiana* / metabolism
  • Plant Roots / metabolism

Substances

  • Antimicrobial Cationic Peptides
  • cecropin A