Successful use of Nicotiana tabacum hairy roots for the recombinant production of Cecropin A peptide
Biotechnol Appl Biochem. 2021 Mar 31. doi: 10.1002/bab.2158. Online ahead of print.
Cecropin A, as an antimicrobial peptide (AMP), is possible to use in medical and agricultural fields as a new and safe bio-control 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 ELISA, respectively. The highest Cecropin A amount was detected in line 4 of the transgenic lines using ELISA in comparison to the non-transgenic 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 antimicrobial peptide 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. This article is protected by copyright. All rights reserved.