Purification and MIC analysis of antimicrobial proteins from Cucumis sativus L. seeds

Abstract

BackgroundCucumis sativus L. (cucumber), from the family Cucurbitaceae, is a therapeutic plant with various pharmacological benefits, broadly utilized as a part of complementary medicine (e.g., Unani, Ayurveda, Siddha, and Traditional Chinese). In light of past research discoveries, this plant had been chosen to consider its potential antibacterial action.MethodsExtracts were purified by dialysis and ion exchange chromatography strategy and then assayed for antibacterial activity against four standard pathogenic bacterial strains known to cause foodborne infections and spoilage of food and herbal drugs. Antimicrobial peptides were extracted from seeds using a sodium phosphate citrate (pH 7.2) - CTAB cradle (pH 6.0).ResultsThe highest protein concentration was seen with elute fractions 1 and 3 (370 mg/mL) compared with elute fractions 2 and 4 (340 mg/mL). Among the bacteria utilized, E. coli was clearly the most sensitive out of selected four strains.ConclusionOur results suggest that Cucumis sativus L seeds extracts have significant potentials as new antimicrobial agents.

Corynebacterium accolens Has Antimicrobial Activity against Staphylococcus aureus and Methicillin-Resistant S. aureus Pathogens Isolated from the Sinonasal Niche of Chronic Rhinosinusitis Patients

“…Inactivation experiments of CFCS were carried out using Proteinase K and heat treatment as previously described [37] with minor amendments. An aliquot of CFCS from selected C. accolens strains were treated with proteinase K (Sigma-Aldrich, St Louis, MO, USA), at a final concentration of 1 mg/mL at 37 °C for 5 h. After incubation, the samples were subjected to heat treatment at 55 °C for 30 min to inactivate protease enzymes.…”

Characterising four Sarconesiopsis magellanica (Diptera: Calliphoridae) larval fat body-derived antimicrobial peptides

“…The plate was incubated for 19 h at 37ºC, read at OD 620 nm wavelength and percentage growth calculated using the following formula: % survival = (450 nm Abs culture medium -450 nm Abs filtrate or fraction extract / 450 nm Abs culture medium -450 nm Abs bacterial suspension) x 100. (26) Minimum inhibitory concentration (MIC) measurements -The assays for determining MIC were started from a concentrated 20 μL solution of each synthetic AMP, followed by dilutions at different concentrations (250, 125, 62.5, 31.25, 15.62, 7.81, 3.90, 1.95 and 0.97 μM). The four bacterial strains had been grown for 18-24 h at 37ºC in 5% CO 2 and then 20 μL of each solutions was taken and mixed with 80 μL bacterial suspension (1.5 x 10 8 CFU/mL) from each selected strain in a 96-well plate; 100 μL Mueller Hinton broth was used as growth control and 20 μL antibiotic mixed with 80 μL bacterial suspension as inhibition control.…”

Antibacterial potential of Antimicrobial peptides containing whole proteins of two Adiantum species from Himalaya against selected human bacterial pathogens

“…125 mM solution of urea in water served the solubilization purpose nicely, hence further high concentrations of urea were not used. (Akeel et al, 2018) also compared two different solubilization buffers for extraction of antimicrobial proteins from seeds of Cucumis sativus L.…”

“…Although they did not study the effect of different buffers on solubilization of proteins for antimicroassay, but the requirement of better protein solubilization buffer was the reason for using two different buffers by (Akeel et al, 2018). Results of antimicrobial activity after solubilization of proteins in SLB and AU are shown in Tables 2 and 3, respectively.…”