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Table 1.

List of phytochemicals selected from B. vulgaris for docking against AChE.

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Table 1 Expand

Fig 1.

Structures of phytochemicals of B. vulgaris a) Ascorbic acid b) Niacin c) Pyridoxine d) Folic acid e) Gallic acid f) Catechol g) p-coumaric acid h) Ferulic acid.

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Fig 1 Expand

Fig 2.

Structures of phytochemicals of B. vulgaris a) o-coumaric acid b) Cinnamic acid c) Myricetin d) Naringenin e) Kaempferol f) Apigenin g) Betanin h) Glycine betaine.

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Fig 2 Expand

Table 2.

Results of phytochemicals examined for Lipinski rule.

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Table 2 Expand

Fig 3.

a) Enzyme ligand interactions within the binding domain of AChE for folic acid (FA) and myricetin (MC) b) Enzyme ligand interactions within the active binding domain of AChE for betanin (BE) and donepezil (DP).

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Fig 3 Expand

Table 3.

Interaction details of phytochemicals in the proposed site of AChE enzyme.

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Table 3 Expand

Fig 4.

Percentage inhibition of AChE activity from 12.5–400 μM concentration of betanin, glycine betaine and the reference standard drug, donepezil.

Results are presented as mean ± SEM for experimental triplicates ****P < 0.0001; **P = 0.0014.

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Fig 4 Expand

Fig 5.

Percentage inhibition of AChE activity at 100 μM concentration of betanin, glycine betaine and the reference standard drug, donepezil.

Results are presented as mean ± SEM for experimental triplicates.

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Fig 5 Expand

Fig 6.

a) Dose response analysis of betanin was performed using serial dilutions (12.5 μM– 400 μM). Nonlinear regression analysis depicted 19.34 μM concentration as IC50. b) Dose response analysis of glycine betaine was performed using serial dilutions (12.5 μM– 400 μM). Nonlinear regression analysis depicted 16.41 μM concentration as IC50.

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Fig 6 Expand

Fig 7.

Dose response analysis of standard reference donepezil was performed using serial dilutions (12.5 μM– 400 μM).

Nonlinear regression analysis depicted 14.27 μM concentration as IC50.

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Fig 7 Expand

Fig 8.

MD simulation interaction diagrams of 20 ns trajectory showing RMSD plot for complex acetyl cholinesterase- Betanin (A), acetyl cholinesterase- Betaine (B), acetyl cholinesterase- Myricetin (C) and acetyl cholinesterase-Donepezil (D) respectively.

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Fig 8 Expand

Fig 9.

Protein Root Mean Square Fluctuation (RMSF) plots (Angstrom).

(A) RMSF trajectory plot of acetyl cholinesterase- Betanine complex showing residue-wise fluctuation, (B) RMSF trajectory plot for acetyl cholinesterase- Betaine complex showing residue-wise fluctuation, (C) RMSF trajectory plot of acetyl cholinesterase with Myricetin complex showing residue-wise fluctuation, (D) RMSF trajectory plot of acetyl cholinesterase with Donepezil complex showing residue-wise fluctuation.

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Fig 9 Expand

Fig 10.

Histogram (stacked bar chart) showing forming H-bonds interactions (green color), hydrophobic interactions (gray violet color), and water bridges (blue color) during 50 ns simulation for complex acetyl cholinesterase-Betanin (A), acetyl cholinesterase- Betaine (B), acetyl cholinesterase- Myricetin (C) and acetyl cholinesterase-Donepezil (D).

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Fig 10 Expand

Fig 11.

The time frame evolution against the radius of gyration (Rg) (A, C, E and G) displayed on left and the SASA plots of docked complexes over 50 ns MD simulation (B, D, F and H) displayed on right.

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Fig 11 Expand

Table 4.

IC50 values of phytochemicals along with reference standard in AChE inhibitory assays.

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Table 4 Expand