Molecular farming of antimicrobial peptides: available platforms and strategies for improving protein biosynthesis using modified virus vectors

Abstract

The constant demand for new antibiotic drugs has driven efforts by the scientific community to prospect for peptides with a broad spectrum of action. In this context, antimicrobial peptides (AMPs) have acquired great scientific importance in recent years due to their ability to possess antimicrobial and immunomodulatory activity. In the last two decades, plants have attracted the interest o f the scientific community and industry as regards their potential as biofactories of heterologous proteins. One of the most promising approaches is the use of viral vectors to maximize the transient expression of drugs in the leaves of the plant Nicotiana benthamiana. recently, the MagnifectionTM expression system was launched. This sophisticated commercial platform allows the assembly of the viral particle in leaf cells and the systemic spread of heterologous protein biosynthesis in green tissues caused by Agrobacterium tumefaciens "gene delivery method". The system also presents increased gene expression levels mediated by potent viral expression machinery. These characteristics allow the mass recovery of heterologous proteins in the leaves of N. benthamiana in 8 to 10 days. This system was highly efficient for the synthesis of different classes of pharmacological proteins and contains enormous potential for the rapid and abundant biosynthesis of AMPs.

Transient Expression of Dengue Virus NS1 Antigen in Nicotiana benthamiana for Use as a Diagnostic Antigen

“…Plants have numerous advantages as a platform for the production of proteins used as diagnostic reagents, vaccines, and drugs, also called plant-made pharmaceuticals (PMPs) (Leite et al, 2019). This platform is easily scalable, requiring only soil, water, and light for the production of a large amount of biomass.…”

“…High levels of protein accumulation can be achieved by targeting the heterologous proteins to the appropriate subcellular compartment (Ahmad et al, 2010). The type of cell compartment in which they are found directly influences the folding, assembling, and posttranslational modifications processes, as well as preventing immediate degradation and interference of the polypeptides with cell metabolism (Ahmad et al, 2010;Leite et al, 2019). Typically, the main compartments to which recombinant proteins are targeted include the apoplast, the vacuole, the endoplasmic reticulum, the chloroplasts, and the cytosol (Pereira el at., 2014;Viegas et al, 2015;Miletic et al, 2016) It has also been shown that adding fusion tags, such as elastinlike polypeptides (ELP), hydrophobins (HFBI), and Zera (an Nproline-rich region of gamma zein) to recombinant proteins increases their levels of accumulation.…”

Plant Viruses in Plant Molecular Pharming: Toward the Use of Enveloped Viruses

“…Plant molecular pharming has materialized as a reliable and cost-effective platform for the production of pharmaceuticals, vaccines, and biobetter products with a number of plant-produced proteins starting to be commercialized (Ratner, 2010; Yusibov et al, 2011). Simultaneously, the development of viral vectors, together with agroinfection, culminated in rapid transient expression systems that successfully express high levels of large and complex pharmaceutical proteins and antimicrobial peptides (Fischer and Emans, 2000; Chen et al, 2013; Peyret and Lomonossoff, 2013; Loh et al, 2017; Leite et al, 2019) and enabled the production of plant-virus based VLPs against chronic and infectious diseases (Hefferon, 2018; Sahithi et al, 2019). However, as with their comparative systems counterparts, plant-virus based VLP(s) are inherently constrained in their ability to present complex antigens and glycoproteins.…”

Antimicrobial peptides (AMPs): A promising class of antimicrobial compounds

“…Plant bioreactors are low‐cost production systems for the synthesis of large quantities of heterologous polypeptides in various organs of plants since they only require soil, water and light (Davies, 2010; Obembe et al, 2011). Cn‐AMP1, clavanin A, Cm AMP‐5 and parigidina‐br1, which have antimicrobial and insecticidal activities, were expressed in high yields in the leaves of the tobacco plant ( Nicotiana benthamiana ; Leite et al, 2018).…”