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

Authors :
Hashemi, Samaneh
Niazi, Ali
Baghizadeh, Amin
Taghizadeh, Mohammad Sadegh
Source :
Biotechnology & Applied Biochemistry; Jun2022, Vol. 69 Issue 3, p876-886, 11p
Publication Year :
2022

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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
08854513
Volume :
69
Issue :
3
Database :
Complementary Index
Journal :
Biotechnology & Applied Biochemistry
Publication Type :
Academic Journal
Accession number :
157816487
Full Text :
https://doi.org/10.1002/bab.2158