In vitro and in silico evaluation of fucosterol from Sargassum horridum as potential human acetylcholinesterase inhibitor.

The fucosterol has been reported numerous biological activities. In this study, the activity in vitro of the fucosterol from Sargassum horridum as potential human acetylcholinesterase inhibitor was evaluated. The structural identification was obtained by nuclear magnetic resonance (NMR) spectroscopy and based on experimental data, we combined docking and molecular dynamics simulations coupled to the molecular-mechanics-generalized-born-surface-area approach to evaluating the structural and energetic basis for the molecular recognition of fucosterol and neostigmine at the binding site of acetylcholinesterase (AChE). In addition, the Lineweaver-Burk plot showed the nature of a non-competitive inhibition. The maximum velocity (Vmax) and the constant of Michaelis-Menten (Km) estimated for fucosterol (0.006 µM) were 0.015 1/Vo (ΔA/h and 6.399 1/[ACh] mM-1, respectively. While, for neostigmine (0.14 µM), the Vmax was 0.022 1/Vo (ΔA/h) and Km of 6.726 1/[ACh] mM-1, these results showed a more effective inhibition by fucosterol respect to neostigmine. Structural analysis revealed that neostigmine reaches the AChE binding site reported elsewhere, whereas fucosterol can act as a no-competitive and competitive acetylcholinesterase inhibitor, in agree with kinetic enzymatic experiments. Binding free energy calculations revealed that fucosterol reaches the acetylcholinesterase binding site with higher affinity than neostigmine, which is according to experimental results. Whereas the per-residue decomposition free energy analysis let us identify crucial residues involved in the molecular recognition of ligands by AChE. Results corroborate the ability of theoretical methods to provide crucial information at the atomic level about energetic and structural differences in the binding interaction and affinity from fucosterol with AChE. Communicated by Ramaswamy H. Sarma.

Screening of extracts of algae from Baja California sur, Mexico as reversers of the antibiotic resistance of some pathogenic bacteria.

BACKGROUND AND OBJECTIVES: Sixty ethanol extracts of marine flora of Baja California Sur (Mexico) were screened to evaluate the reversing effect of the bacterial resistance to antibiotics in combination with a sublethal concentration of ampicillin or erythromycin. MATERIALS AND METHODS: The activity was assayed by using a modification of the classical agar-diffusion method against 3 resistant, pathogenic bacteria; Escherichia coil (ATCC BAA196), Staphylococcus aureus (ATCC BAA42), and Streptococcus pyogenes (ATCC BAA946). RESULTS: From the 60 ethanolic extracts, 12 (20%) of them in combination with ampicillin were able to reverse the resistance of Staphylococcus aureus and 8 (13%) with erythromycin yielded the same reversal with Streptococcus pyogenes. An extract from Sargassum horridum was the only one that reversed the resistance to antibiotics against both Staphylococcus aureus and Streptococcus pyogenes. CONCLUSIONS: Our findings suggest that some algae may be source of compounds with the potential to reverse the antibiotic resistance of some bacteria. In addition, of the assayed extracts, 35 (57%) showed inhibitory activity against Staphylococcus aureus, 48 (78%) were active against Streptococcus pyogenes, but none was active against Escherichia coil. The most active extracts were from Laurencia spp., Gelidium robustum, Chnoospora implexa, Padina mexicana, Gracilaria subsecundata, and Dictyopteris undulata.