Functional characterization of a potent antimicrobial and insecticidal chitin binding protein from seeds of Iberis umbellata L.

Saeed, Ahsan; Rafiq, Zahra; Saeed, Qamar; Khaliq, Binish; Ullah, Anwar; Mehmood, Sohaib; Ali, Zahid; Ashraf, Muhammad; Akrem, Ahmed

doi:10.30848/pjb2021-4(16)

Cited by 4 publications

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“…Similar to other sugar-binding proteins like lectins, chitin-binding proteins and chitinases, vicilins also bind to chitin moieties and inhibit fungal and insect growth. 3 - 5 However, the real-time picture of in-vivo molecular interactions between vicilin and chitin molecules is lacking and remains poorly understood. Chitin is an unbranched polymer of amino sugar occurring in the fungal cell wall, in the exoskeleton of invertebrates, peritrophic membrane (PM) and peritrophic gel of the insect gut.…”

Section: Introductionmentioning

confidence: 99%

In-silico Studies Calculated a New Chitin Oligomer Binding Site Inside Vicilin: A Potent Antifungal and Insecticidal Agent

et al. 2022

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Vicilins are major seed storage proteins and show differential binding affinities toward sugar moieties of fungal cell wall and insect gut epithelium. Hence, purpose of study is the thorough in-silico characterization of interactions between vicilin and chitin oligomer followed by fungal and insecticidal bioassays. This work covers the molecular simulation studies explaining the interactions between Pisum sativum vicilin ( PsV) and chitin oligomer followed by protein bioassay against different pathogens. LC-MS/MS of purified PsV (∼50 kDa) generated residual data along high pea vicilin homology (UniProtKB ID; P13918). Predicted model ( PsV) indicated the characteristic homotrimer joined through head-to-tail association and each monomer is containing a bicupin domain. PsV site map analysis showed a new site (Site 4) into which molecular docking confirmed the strong binding of chitin oligomer (GlcNAc)4. Molecular dynamics simulation data (50 ns) indicated that chitin-binding site was comprised of 8 residues (DKEDRNEN). However, aspartate and glutamate significantly contributed in the stability of ligand binding. Computational findings were further verified via significant growth inhibition of Aspergillus flavus, A. niger, and Fusarium oxysporum against PsV. Additionally, the substantial adult population of Brevicoryne brassicae was reduced and different life stages of Tribolium castaneum also showed significant mortality.

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Section: Introductionmentioning

confidence: 99%

In-silico Studies Calculated a New Chitin Oligomer Binding Site Inside Vicilin: A Potent Antifungal and Insecticidal Agent

et al. 2022

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“…It was also observed that another chitin‐binding protein called Iu‐CBP was isolated from Iberis umbellata seeds, and in addition to demonstrating activity against several bacteria, it also showed activity against the fungus Aspergillus flavus . [ 62 ] In addition to the families reported above showing activity against phytopathogenic yeasts and fungi, [ 19 ] other chitin‐binding proteins, such as lectins, vicilins and chitinases, have been reported to be toxic to insects. [ 16,29,63 ]…”

Section: Resultsmentioning

confidence: 99%

Chitin‐binding peptides from Capsicum annuum with antifungal activity and low toxicity to mammalian cells and Galleria mellonella larvae

Gonçalves,

Silva,

dos Santos

et al. 2024

Peptide Science

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In recent years, there have been several reports of the presence of toxic proteins in cultivated or wild plant species, which are implicated in plant defense mechanisms. The existence of these proteins raises the possibility of biotechnological applications originating from the development of new techniques to combat diseases caused by fungi. In this context, there are chitin‐binding proteins. Chitin is an essential component of the fungal cell wall, so chitin‐binding proteins are important in controlling fungal growth. Thus, the objective of this study was to characterize and evaluate the in vitro antimicrobial effect of peptides with chitin binding properties isolated from Capsicum annuum seeds on the growth of the genus Candida. Initially, proteins were extracted in phosphate pH 5.4, and a chitin column was equilibrated with sodium acetate (0.08 M, pH 4.5), where 50 mg of the peptide‐rich heated fraction from each species was applied. Subsequently, the retained material was eluted with 0.1 M HCl. Tricine SDS–PAGE was used to visualize the peptides. After chromatography, two fractions, F1 (not retained in the chitin column) and F2 (retained in the chitin column, named Ca‐F2), were obtained. Electrophoresis showed major protein bands between 3 and 14 kDa. Electrophoresis from chitin affinity chromatography also showed major bands between 3 and 14 kDa, especially for Ca‐F2 retained in the column. One peptide obtained from the F2 fraction was identified by mass spectrometry and showed similarity to seed 2S albumin, named Ca‐Alb2S. Ca‐F2 inhibited the growth of C. albicans and C. tropicalis, was not toxic to mammalian cells and still had a high survival rate when tested in vivo on Galleria mellonella larvae. This is the first report of chitin‐binding peptides isolated from Capsicum seeds through an affinity column and their biological activities. These studies are at an early stage; therefore, other tests are needed to study the mechanism of action of the fraction, since the findings indicate great potential for the development of new antifungal molecules.

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“…Since chitin-binding proteins (vicilin) contain abundant positively charged residues (such as arginine), they could interact with the negatively charged components of the cell membrane, causing membrane disruption (depolarization of lipid membranes) and cell lysis (Figure S4) [ 64 ]. It is generally known that chitin found in fungal cell walls interacts with a group of proteins known as chitin binding proteins [ 20 , 53 ], causing growth inhibition in these species, as shown in [ 44 , 65 ]. These findings suggest that legume seed vicilins may interact with organisms that contain glucose, sucrose, chitin, chitin derivatives, or N -acetylglucosamine-containing glycoconjugates in structures exposed to the outside environment [ 66 ].…”

Section: Discussionmentioning

confidence: 99%

“…Survival data was noted after every 3 days up to 9 days and number of surviving adults were noted and compared with control [ 26 ] In the second batch, the rice kernels having the progeny of adults were taken and hatched at an optimum temperature of 27 °C for 35 days of incubation time with the same contaminated rice kernels. The surviving individuals were left for the next progeny to see the protein effectivity on the lifespan of rice weevils after 35 (F1 generation) and 65 (F2 generation) days [ 65 ].…”

Section: Methodsmentioning

confidence: 99%

In Silico Analysis and Functional Characterization of Antimicrobial and Insecticidal Vicilin from Moth Bean (Vigna aconitifolia (Jacq.) Marechal) Seeds

et al. 2022

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Vicilin has nutraceutical potential and different noteworthy medicative health-promoting biotic diversions, and it is remarkable against pathogenic microorganisms and insects. In this study, Vigna aconitifolia vicilin (VacV) has been identified and characterized from the seed of Vigna aconitifolia (Jacq.) Marechal (Moth beans). LC-MS/MS analysis of VacV provided seven random fragmented sequences comprising 238 residues, showing significant homology with already reported Vigna radiata vicilin (VraV). VacV was purified using ammonium sulfate precipitation (60%) followed by size exclusion chromatography on Hi-Load 16/60 Superdex 200 pg column and anion-exchange chromatography (Hi trap Q FF column). Purified VacV showed a major ~50 kDa band and multiple lower bands on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under both reduced and non-reduced conditions. After all, a three-dimensional molecular structure of VacV was predicted, which showed β-sheeted molecular conformation similar to crystallographic structure of VraV. All Vicilins from V. aconitifolia and other plants were divided into six sub-groups by phylogenetic analysis, and VacV shared a high degree of similarity with vicilins of Vigna radiata, Pisum sativum, Lupinus albus, Cicer arietinum and Glycine max. Additionally, VacV (20 μg) has significant growth inhibition against different pathogenic bacteria along strong antifungal activity (50 μg). Likewise, VacV (3.0 mg) produced significant growth reduction in Rice Weevil Sitophilus oryzae larvae after 9 days compared with control. Furthermore, by using MMT assay, the cytotoxicity effect of VacV on the growth of HepG2 liver cancerous cells was tested. VacV showed cytotoxicity against the HepG-2 line and the acquired value was 180 µg after 48 h. Finally, we performed molecular docking against caspase-3 protein (PDB ID: 3DEI) for VacV bioactive receptor interface residues. Hence, our results reveal that VacV, has nutraceutical potential and moth beans can be used as a rich resource of functional foods.

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Functional characterization of a potent antimicrobial and insecticidal chitin binding protein from seeds of Iberis umbellata L.

Cited by 4 publications

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In-silico Studies Calculated a New Chitin Oligomer Binding Site Inside Vicilin: A Potent Antifungal and Insecticidal Agent

In-silico Studies Calculated a New Chitin Oligomer Binding Site Inside Vicilin: A Potent Antifungal and Insecticidal Agent

Chitin‐binding peptides from Capsicum annuum with antifungal activity and low toxicity to mammalian cells and Galleria mellonella larvae

In Silico Analysis and Functional Characterization of Antimicrobial and Insecticidal Vicilin from Moth Bean (Vigna aconitifolia (Jacq.) Marechal) Seeds

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