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. 2020 Jul 31;25(15):3478.
doi: 10.3390/molecules25153478.

Fractionation of Biomolecules in Withania coagulans Extract for Bioreductive Nanoparticle Synthesis, Antifungal and Biofilm Activity

Murtaza Hasan  1   2 , Ayesha Zafar  2 , Irum Shahzadi  2 , Fan Luo  1 , Shahbaz Gul Hassan  3 , Tuba Tariq  2 , Sadaf Zehra  4 , Tauseef Munawar  5 , Faisal Iqbal  5 , Xugang Shu  1
Affiliations

Fractionation of Biomolecules in Withania coagulans Extract for Bioreductive Nanoparticle Synthesis, Antifungal and Biofilm Activity

Murtaza Hasan et al. Molecules. .

Abstract

Withania coagulans contains a complex mixture of various bioactive compounds. In order to reduce the complexity of the plant extract to purify its phytochemical biomolecules, a novel fractionation strategy using different solvent combination ratios was applied to isolate twelve bioactive fractions. These fractions were tested for activity in the biogenic synthesis of cobalt oxide nanoparticles, biofilm and antifungal activities. The results revealed that plant extract with bioactive fractions in 30% ratio for all solvent combinations showed more potent bioreducing power, according to the observed color changes and the appearance of representative absorption peaks at 500-510 nm in the UV-visible spectra which confirm the synthesis of cobalt oxide nanoparticles (Co3O4 NPs). XRD diffraction was used to define the crystal structure, size and phase composition of the products. The fractions obtained using 90% methanol/hexane and 30% methanol/hexane showed more effectiveness against biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus so these fractions could potentially be used to treat bacterial infections. The 90% hexane/H2O fraction showed excellent antifungal activity against Aspergillus niger and Candida albicans, while the 70% methanol/hexane fraction showed good antifungal activity for C. albicans, so these fractions are potentially useful for the treatment of various fungal infections. On the whole it was concluded that fractionation based on effective combinations of methanol/hexane was useful to investigate and study bioactive compounds, and the active compounds from these fractions may be further purified and tested in various clinical trials.

Keywords: antibiofilm; biomolecules; fractionation; microbial infection; reducing activity.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic illustration of W. coagulans biomolecules and their applications.
Figure 2
Figure 2
Bioreducing potential of W. coagulans based on: (a) methanol (b) hexane (c) methanol/hexane (mixture) fractions for Co3O4 NPs synthesis.
Figure 3
Figure 3
Bioreducing potential of Withania coagulans based on methanol, hexane and methanol/hexane (mixtures) using (a) 30% fraction, (b) 50% fraction, (c) 70% fraction, (d) 90% fraction.
Figure 4
Figure 4
XRD analysis of Co3O4 nanoparticles based on solvent fractions: (a) methanol fraction (b) hexane fraction, (c) methanol/hexane fraction.
Figure 5
Figure 5
Variation in Co3O4 nanoparticle shape based on solvent fractions: (a) methanol fraction (b) hexane fraction, (c) methanol/hexane fraction.
Figure 6
Figure 6
Antibiotic selectivity of (a) clindamycin, moxifloxacin (b) penicillin, ciprofloxacin against P. aeruginosa (c) penicillin, ciprofloxacin (d) clindamycin, moxifloxacin against S. aureus.
Figure 7
Figure 7
Biofilm activity of W. coagulans 12 fraction against P. aeruginosa with concentration (a) 5 mg/mL (b) 10 mg/mL.
Figure 8
Figure 8
Biofilm activity of W. coagulans 12 fraction against S. aureus with concentration (a) 5 mg/mL (b) 10 mg/mL.
Figure 9
Figure 9
Antifungal activity of 12 W. coagulans fractions against A. niger measured by zone of inhibition: (a) Positive control; (b) negative control; (c) methanol fractions; (d) hexane fraction 30%, 50%; (e) hexane fraction 70%, 90%; (f) methanol/hexane fraction 30%, 50% (g); methanol/hexane fraction 70%, 90% (h).
Figure 10
Figure 10
Antifungal activity of 12 W. coagulans fractions against C. albicans (zone of inhibition): (a) Positive control (b) negative control (c) methanol fractions (d) hexane fraction 30%, 50% (e) hexane fraction 70%, 90% (f) methanol/hexane fraction 30%, 50% (g) methanol/hexane fraction 70%, 90% (h).
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