Ligand-Induced structural changes in TEM‑1 probed by molecular dynamics and relative binding free energy calculations

The TEM family of enzymes has had a crucial impact on the pharmaceutical industry due to their important role in antibiotic resistance. Even with the latest technologies in structural biology and genomics, no 3D structure of a TEM- 1/antibiotic complex is known previous to acylation. Therefore, the comprehension of their capability in acylate antibiotics is based on the protein macromolecular structure uncomplexed. In this work, molecular docking, molecular dynamic simulations, and relative free energy calculations were applied in order to get a comprehensive and thorough analysis of TEM-1/ampicillin and TEM-1/amoxicillin complexes. We described the complexes and analyzed the effect of ligand binding on the overall structure. We clearly demonstrate that the key residues involved in the stability of the ligand (hot-spots) vary with the nature of the ligand. Structural effects such as (i) the distances between interfacial residues (Ser70−Oγ and Lys73−Nζ, Lys73−Nζ and Ser130−Oγ, and Ser70−Oγ−Ser130−Oγ), (ii) side chain rotamer variation (Tyr105 and Glu240), and (iii) the presence of conserved waters can be also influenced by ligand binding. This study supports the hypothesis that TEM-1 suffers structural modifications upon ligand binding.

Convergence to the European Energy Policy in European countries: case studies and comparison

Purpose – Our paper aims at analyzing how different European countries cope with the European Energy Policy, which proposes a set of measures (free energy market, smart meters, energy certificates) to improve energy utilization and management in Europe. Design/methodology/approach – The paper first reports the general vision, regulations and goals set up by Europe to implement the European Energy Policy. Later on, it performs an analysis of how some European countries are coping with the goals, with financial, legal, economical and regulatory measures. Finally, the paper draws a comparison between the countries to present a view on how Europe is responding to the emerging energy emergency of the modern world. Findings – Our analysis on different use cases (countries) showed that European countries are converging to a common energy policy, even though some countries appear to be later than others In particular, Southern European countries were slowed down by the world financial and economical crisis. Still, it appears that contingency plans were put into action, and Europe as a whole is proceeding steadily towards the common vision. Research limitations/implications – European countries are applying yet more cuts to financing green technologies, and it is not possible to predict clearly how each country will evolve its support to the European energy policy. Practical implications – Different countries applied the concepts and measures in different ways. The implementation of the European energy policy has to cope with the resulting plethora of regulations, and a company proposing enhancement regarding energy management still has to possess robust knowledge of the single country, before being able to export experience and know-how between European countries. Originality/Value – Even though a few surveys on energy measures in Europe are already part of the state-of-the-art, organic analysis diagonal to the different topics of the European Energy Policy is missing. Moreover, this paper highlights how European countries are converging on a common view, and provides some details on the differences between the countries, thus facilitating parties interesting into cross-country export of experience and technology for energy management.

Microencapsulation of herbicide MCPA with native beta-cyclodextrin and its methyl and hydroxypropyl derivatives: An experimental and theoretical investigation

When a pesticide is released into the environment, most of it is lost before it reaches its target. An effective way to reduce environmental losses of pesticides is by using controlled release technology. Microencapsulation becomes a promising technique for the production of controlled release agricultural formulations. In this work, the microencapsulation of chlorophenoxy herbicide MCPA with native b-cyclodextrin and its methyl and hydroxypropyl derivatives was investigated. The phase solubility study showed that both native and b-CD derivatives increased the water solubility of the herbicide and inclusion complexes are formed in a stoichiometric ratio of 1:1. The stability constants describing the extent of formation of the complexes have been determined by phase solubility studies. 1H NMR experiments were also accomplished for the prepared solid systems and the data gathered confirm the formation of the inclusion complexes. 1H NMR data obtained for the MCPA/CDs complexes disclosed noticeable proton shift displacements for OCH2 group and H6 aromatic proton of MCPA provided clear evidence of inclusion complexation process, suggesting that the phenyl moiety of the herbicide was included in the hydrophobic cavity of CDs. Free energy molecular mechanics calculations confirm all these findings. The gathered results can be regarded as an essential step to the development of controlled release agricultural formulations containing herbicide MCPA.