Controlled oxidation of hydrocarbons by the membrane-bound methane monooxygenase: the case for a tricopper cluster

[Figure: see text]. The growing need for inexpensive methods to convert methane to methanol has sparked considerable interest in methods that catalyze this process. The integral membrane protein particulate methane monooxygenase (pMMO) mediates the facile conversion of methane to methanol in methanotrophic bacteria. Most evidence indicates that pMMO is a multicopper enzyme, and these copper ions support redox, dioxygen, and oxo-transfer chemistry. However, the exact identity of the copper species that mediates the oxo-transfer chemistry remains an area of intense debate. This highly complex enzyme is notoriously difficult to purify because of its instability outside the lipid bilayer and tendency to lose its essential metal cofactors. For this reason, pMMO has resisted both initial identification and subsequent isolation and purification for biochemical and biophysical characterization. In this Account, we describe evidence that pMMO is a multicopper protein. Its unique trinuclear copper cluster mediates dioxygen chemistry and O-atom transfer during alkane hydroxylation. Although a recent crystal structure did not show this tricopper cluster, we provide compelling evidence for such a cluster through redox potentiometry and EPR experiments on the "holo" enzyme in pMMO-enriched membranes. We also identify a site in the structure of pMMO that could accommodate this cluster. A hydrophobic pocket capable of harboring pentane, the enzyme's largest known substrate, lies adjacent to this site. In addition, we have designed and synthesized model tricopper clusters to provide further chemical evidence that a tricopper cluster mediates the enzyme's oxo-transfer chemistry. These biomimetic models exhibit similar spectroscopic properties and chemical reactivity to the putative tricopper cluster in pMMO. Based on computational analysis using density functional theory (DFT), triangular tricopper clusters are capable of harnessing a "singlet oxene" upon activation by dioxygen. An oxygen atom is then inserted via a concerted process into the C-H bond of an alkane in the transition state during hydroxylation. The turnover frequency and kinetic isotope effect predicted by DFT show excellent agreement with experimental data.     

The Multifunctional Sactipeptide Ruminococcin C1 Displays Potent Antibacterial Activity In Vivo as Well as Other Beneficial Properties for Human Health

The world is on the verge of a major antibiotic crisis as the emergence of resistant bacteria is increasing, and very few novel molecules have been discovered since the 1960s. In this context, scientists have been exploring alternatives to conventional antibiotics, such as ribosomally synthesized and post-translationally modified peptides (RiPPs). Interestingly, the highly potent in vitro antibacterial activity and safety of ruminococcin C1, a recently discovered RiPP belonging to the sactipeptide subclass, has been demonstrated. The present results show that ruminococcin C1 is efficient at curing infection and at protecting challenged mice from Clostridium perfringens with a lower dose than the conventional antibiotic vancomycin. Moreover, antimicrobial peptide (AMP) is also effective against this pathogen in the complex microbial community of the gut environment, with a selective impact on a few bacterial genera, while maintaining a global homeostasis of the microbiome. In addition, ruminococcin C1 exhibits other biological activities that could be beneficial for human health, as well as other fields of applications. Overall, this study, by using an in vivo infection approach, confirms the antimicrobial clinical potential and highlights the multiple functional properties of ruminococcin C1, thus extending its therapeutic interest.     

Challenges for plasma etch integration of ferroelectric capacitors in FeRAM's and DRAM's

Abstract

The plasma etch process requirements are different for etching 2μm ferroelectric capacitor structures in FeRAM's (SRAM) vs. the smaller capacitor sizes (0.2–0.5 μm) of DRAM's. Plasma etch integration of ferroelectric capacitors presents three major differences between FeRAM's and DRAM's. The first difference is in the ferroelectric capacitor structure. FeRAM's use planar capacitors with top side metal contacts to vias while DRAM's use vertical capacitor structures with bottom side contact to a poly post structure. The second major difference is in material selected and thickness of layers. FeRAM's use thicker electrodes of Pt or Ir and a thicker PZT or Y1 dielectric layer. FeRAM's use a thick bottom electrode (and a thin top electrode) consisting of Pt, Ru or Ir and a thin BST dielectric layer. The third major difference is the plasma etch process requirements for the two devices. FeRAM's require a clean etch process and no corrosion. Profile is not critical but should be maintained at greater than 60° for 2μm bottom post electrode. An HRe^? (Highly Density Reflected Electron) etch system is used to develop process trends for ferroelectric capacitor applications.     

Elucidation of structural difference in theaflavins for modulation of starch digestion

The relationship between structure and activity of theaflavins against human pancreatic α-Amylase was investigated by in vitro and in silico methods. The IC₅₀ and total energy value showed that inhibitory effects followed the order: theaflavin-3, 3’-di-O-gallate > theaflavin-3’-O-gallate > theaflavin-3-O-gallate > theaflavin. Inhibitory activity was depended on hydroxyl groups and galloyl moieties of theaflavins to interact with the catalytic residues of the active site of α-Amylase by hydrogen bonds and π–π (aromatic–aromatic) interactions. The galloylated theaflavin has higher binding affinity with α-Amylase than non-galloylated theaflavin. The study showed that theaflavins might act as natural enzyme inhibitors with potential health benefits, which provide a foundation for designing novel functional food for effective controlling of starch digestion and postprandial glucose levels.     

Experimental study of the burner for FAA fire test: NexGen burner

The NexGen (Sonic) burner is the new burner developed by the Federal Aviation Administration, FAA, to replace old oil burners used for the required fire certification tests on power plant‐related materials, as it provides the capability to control both air and fuel flow rates. During a fire test, the burner is supposed to simulate a certain fire condition, so the flame properties should be robust and repeatable. The NexGen burner can achieve this due to the precise fuel and air controls. However, the current calibration criterion (ISO2685:1998 and AC20‐135) may not be good enough to ensure consistent flame properties. In the presented work, the sensitivity of the burner performance to air and fuel flow rate, as measured by the temperature and heat flux for calibration purposes, was studied. Additionally, the influence of the turbulator and the thermocouple size used for flame calibration was also studied. The impact of varying fuel/air ratio and thermocouple sizes was studied by conducting fire tests on aluminum samples, to show the inadequacies in the current calibration standards.     

Three-dimensional distinct element modelling and dynamic runout analysis of a landslide in gneissic rock, British Columbia, Canada

The McAuley Creek Landslide is a 6?million m³ gneissic rock slope failure that occurred in British Columbia (Canada) in late May–early June 2002. The geological strength index was used to characterize the quality of the overall rock mass and its reduced (damaged) quality near tectonic structures and alteration zones. Potential slope failure mechanisms were investigated using four analysis techniques including: kinematic analysis, surface wedge limit equilibrium (combination) analysis, block theory and three-dimensional distinct element models. Results from all four analyses suggested that the dominant slope failure mechanism was wedge sliding along the intersection of the gneissic foliation and a steeply dipping discontinuity set striking perpendicular to the slope. Of the 6?million m³ of material involved in the landslide, an estimated 5 million?m³ was deposited immediately below the source area against the opposite valley wall, with the remaining 1 million m³ travelling an additional 1.6?km downstream. The runout behaviour was investigated using a three-dimensional dynamic analysis code.