Molecular Identification and Characterization of Some Knottin Antimicrobial Peptides From White Mustard (Sinapis alba L.) Plant

Bahramnejad, Nastaran; Sohrabi, Seyyed Mohsen; Mohammadi, Mohsen

doi:10.22034/pgr.2025.2082488.1028

Molecular Identification and Characterization of Some Knottin Antimicrobial Peptides From White Mustard (Sinapis alba L.) Plant

Document Type : Original Article

Authors

Nastaran Bahramnejad ¹

Seyyed Mohsen Sohrabi ²

Mohsen Mohammadi ¹

¹ Department Pharmacognosy and Pharmaceutical Biotechnology, Faculty of Pharmacy, Lorestan University of Medical Sciences, Khorramabd, Iran

² Department of Production Engineering and Plant Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

10.22034/pgr.2025.2082488.1028

Abstract

Antimicrobial peptides (AMPs) are a diverse group of low-molecular-weight bioactive molecules that serve as the primary defense line of the innate immune system against infections in all multicellular organisms. In plants, these peptides exhibit high biodiversity and belong to several families, being produced both constitutive and inducible in response to environmental stresses. Knottin antimicrobial peptides are a class of peptides found in plants, animals, and insects which are cysteine-rich peptides. This study aimed to identify and characterize some Knottin antimicrobial peptides from white mustard (Sinapis alba L.). In this research, the gene sequences of seven Knottin antimicrobial peptides from white mustard were predicted and identified using bioinformatics and laboratory techniques. Initially, bioinformatics methods were used to predict the encoding sequences of the target peptides from the transcriptome of white mustard. Subsequently, these encoding sequences were isolated and identified using polymerase chain reaction (PCR). The PCR products were sequenced and analyzed using various bioinformatics tools. The physicochemical properties of the seven identified Knottin peptides from white mustard were predicted, with molecular weights ranging from 9.92 to 11.05 kDa, isoelectric pH values from 4.46 to 7.89, instability index from 38.06 to 63.9, aliphatic index from 62.36 to 81.82, and GRAVY values ranging from -0.249 to 0.18. It was also found that the Knottin peptides of white mustard contain four intramolecular disulfide bonds formed by eight conserved cysteine residues, contributing to their unique structure and stability. Additionally, bioinformatics analysis revealed that all the identified Knottin peptides in white mustard have potentially antimicrobial activities. Given that plants produce a wide range of antimicrobial peptides, and considering that purification methods can be complex, costly, and time-consuming, these peptides can be identified, designed, and chemically synthesized or produced recombinantly using bioinformatics tools and after validation through further experiments, could be a promising therapeutic potential to develop as new antimicrobial agents against drug-resistant pathogens.

Keywords

Bioinformatics

Anti-microbial peptides

Sinapis alba L

Knottin

PCR

Subjects

Bioinformatics

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