Abstract
Resistance to common drugs by microorganisms and cancers has become a major issue in modern healthcare, increasing the number of deaths worldwide. Novel therapeutic agents with a higher efficiency and less side effects for the treatment of certain diseases are urgently needed. Plant defensins have an integral role in a hosts’ immune system and are attractive candidates for combatting drug-resistant microorganisms. Interestingly, some of these defensins also showed great potential due to their cytotoxic activity toward cancer cells. In this study, a defensin encoding gene was isolated from five legume seeds using 3′ rapid amplification of cDNA ends (3′ RACE) with degenerate primers and cDNA cloning strategies. Bioinformatic tools were used for in silico identification and the characterization of new sequences. To study the functional characteristics of these unique defensins, the gene encoded for Sesbania javanica defensin, designated as javanicin, was cloned into pTXB-1 plasmid and expressed in the Escherichia coli Origami 2 (DE3) strain. Under optimized conditions, a 34-kDa javanicin-intein fusion protein was expressed and approximately 2.5–3.5 mg/L of soluble recombinant javanicin was successfully extracted with over 90% purity. Recombinant javanicin displayed antifungal properties against human pathogenic fungi, including resistant strains, as well as cytotoxic activities toward the human breast cancer cell lines, MCF-7 & MDA-MB-231. Recombinant javanicin holds great promise as a novel therapeutic agent for further medical applications.
Highlights
Resistance to common drugs by microorganisms and cancers has become a major issue in modern healthcare, increasing the number of deaths worldwide
The nucleotide and deduced amino acid sequences of these unique plant defensins from the seeds of V. mungo, C. juncea, L. purpureus, S. javanica and C. gladiata were recorded in GenBank accession No MH045506-MH045510, respectively
The results indicated that these defensin antimicrobial peptides were highly conserved with a 75-amino acids pro-peptide consisting of a 28 amino acids signal sequence analyzed by SignalP 4.1 and the C-terminal 47 residues mature peptide
Summary
Resistance to common drugs by microorganisms and cancers has become a major issue in modern healthcare, increasing the number of deaths worldwide. Plant defensins have an integral role in a hosts’ immune system and are attractive candidates for combatting drugresistant microorganisms Some of these defensins showed great potential due to their cytotoxic activity toward cancer cells. Unique therapeutic agents with higher efficiency and decreased side effects for the treatment of these hard to cure diseases are urgently needed Regarding their multifaceted effects, antimicrobial peptides (AMPs), a natural host defense peptide, are one of the more attractive agents for the treatment of these issues, which are capable combatting both drug-sensitive and drug-resistant microbial pathogens and exhibit antiviral and anticancer activities. AMPs induce modulatory functions in the innate immune response[6] These peptides exact mode of action towards cancer cells is still unclear, several modes of function on microorganisms have. Fusion partners are capable of protecting AMPs from proteolytic cleavage, improving the purification process, increasing the expression efficiency and improving the solubility of recombinant peptides[18]
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