Molecular Basis of C–N Bond Cleavage by the Glycyl Radical Enzyme Choline Trimethylamine-Lyase

Deamination of choline catalyzed by the glycyl radical enzyme choline trimethylamine-lyase (CutC) has emerged as an important route for the production of trimethylamine, a microbial metabolite associated with both human disease and biological methane production. Here, we have determined five high-resolution X-ray structures of wild-type CutC and mechanistically informative mutants in the presence of choline. Within an unexpectedly polar active site, CutC orients choline through hydrogen bonding with a putative general base, and through close interactions between phenolic and carboxylate oxygen atoms of the protein scaffold and the polarized methyl groups of the trimethylammonium moiety. These structural data, along with biochemical analysis of active site mutants, support a mechanism that involves direct elimination of trimethylamine. This work broadens our understanding of radical-based enzyme catalysis and will aid in the rational design of inhibitors of bacterial trimethylamine production.National Science Foundation (U.S.) (Grant 0645960

A Standard System of Characterization for Olefin Metathesis Catalysts

The success of olefin metathesis has spurred the intense investigation of new catalysts for this transformation. With the development of many different catalysts, however, it becomes increasingly difficult to compare their efficiencies. In this article we introduce a set of six reactions with specific reaction conditions to establish a standard for catalyst comparison in olefin metathesis. The reactions were selected on the basis of their ability to provide a maximum amount of information describing catalyst activity, stability, and selectivity, while being operationally simple. Seven of the most widely used ruthenium-based olefin metathesis catalysts were evaluated with these standard screens. This standard is a useful tool for the comparison and evaluation of new metathesis catalysts

The Potential of Water Saving and Water Capturing Innovations: A Case Study of Albuquerque Single Family Homes

The need for innovative water saving and water capturing strategies in Albuquerque single family homes (SFH) is evident by the citys vulnerability to drought, climate change and population growth. The intrinsic value of modern innovations is that they offer the potential to produce the largest, most cost effective and environmentally sound alternative source of water required to meet current and future demand. Moreover, recognizing the intimate relationship between water use and the energy consumed in conveyance, treatment, end uses and waste treatment, allows policy makers to meet water and energy use reduction goals simultaneously, as well as enhance water and energy security opportunity for current and future populations. Three innovations were examined to demonstrate the potential water, energy and monetary savings possible for the Albuquerque Bernalillo County Water Utility Department and its single family home customers. The usage of these three innovations was projected to 2030 and their associated benefits discussed with regard to their potential for generating future opportunity. Two in-home innovations, the dual-flush toilet and the Shower Water Conservation System (SWCS), the latter an innovation designed by the author of this report, Andrew Funk, as well as one exterior innovation, rain water harvesting, were analyzed for their potential alternative source water production, and energy and monetary savings. Using water saving and water capturing innovations in Albuquerque Bernalillo County Water Utility Department single family homes offers access to a significant volume of least cost alternative source water. These types of innovations empower the water utility and its customers to use water and energy resources more efficiently, save money and further decrease Albuquerque\u27s reliance on groundwater, better equipping the City to manage future drought, climate change and population growth

Warming of the Indian Ocean Threatens Eastern and Southern Africa, but could be Mitigated by Agricultural Development

Since 1980, the number of undernourished people in eastern and southern Africa has more than doubled. Rural development stalled and rural poverty expanded during the 1990s. Population growth remains very high and declining per capita agricultural capacity retards progress towards Millennium Development goals. Analyses of in situ station data and satellite observations of precipitation identify another problematic trend. Main growing season rainfall receipts have diminished by approximately 15% in food insecure countries clustered along the western rim of the Indian Ocean. Occurring during the main growing seasons in poor countries dependent on rain fed agriculture, these declines are societally dangerous. Will they persist or intensify? Tracing moisture deficits upstream to an anthropogenically warming Indian Ocean leads us to conclude that further rainfall declines are likely. We present analyses suggesting that warming in the central Indian Ocean disrupts onshore moisture transports, reducing continental rainfall. Thus late 20th century anthropogenic Indian Ocean warming has probably already produced societally dangerous climate change by creating drought and social disruption in some of the world's most fragile food economies. We quantify the potential impacts of the observed precipitation and agricultural capacity trends by modeling millions of undernourished people as a function of rainfall, population, cultivated area, seed and fertilizer use. Persistence of current tendencies may result in a 50% increase in undernourished people. On the other hand, modest increases in per capita agricultural productivity could more than offset the observed precipitation declines. Investing in agricultural development can help mitigate climate change while decreasing rural poverty and vulnerability

The class III ribonucleotide reductase from Neisseria bacilliformis can utilize thioredoxin as a reductant

The class III anaerobic ribonucleotide reductases (RNRs) studied to date couple the reduction of ribonucleotides to deoxynucleotides with the oxidation of formate to CO[subscript 2]. Here we report the cloning and heterologous expression of the Neisseria bacilliformis class III RNR and show that it can catalyze nucleotide reduction using the ubiquitous thioredoxin/thioredoxin reductase/NADPH system. We present a structural model based on a crystal structure of the homologous Thermotoga maritima class III RNR, showing its architecture and the position of conserved residues in the active site. Phylogenetic studies suggest that this form of class III RNR is present in bacteria and archaea that carry out diverse types of anaerobic metabolism.Singapore. Agency for Science, Technology and ResearchNational Science Foundation (U.S.). Graduate Research Fellowship Program (Grant 0645960)United States. Dept. of Energy. Office of Basic Energy Sciences (Contract DE-AC02-06CH11357)National Institutes of Health (U.S.) (Grant GM29595

Biophysical Characterization of Fluorotyrosine Probes Site-Specifically Incorporated into Enzymes:

Fluorinated tyrosines (F[subscript n]Y's, n = 2 and 3) have been site-specifically incorporated into E. coli class Ia ribonucleotide reductase (RNR) using the recently evolved M. jannaschii Y-tRNA synthetase/tRNA pair. Class Ia RNRs require four redox active Y's, a stable Y radical (Y·) in the β subunit (position 122 in E. coli), and three transiently oxidized Y's (356 in β and 731 and 730 in α) to initiate the radical-dependent nucleotide reduction process. F[subscript n]Y (3,5; 2,3; 2,3,5; and 2,3,6) incorporation in place of Y₁₂₂-β and the X-ray structures of each resulting β with a diferric cluster are reported and compared with wt-β2 crystallized under the same conditions. The essential diferric-F[subscript n]Y· cofactor is self-assembled from apo F[subscript n]Y-β2, Fe ²⁺, and O₂ to produce ∼1 Y·/β2 and ∼3 Fe ³⁺ /β2. The F[subscript n]Y· are stable and active in nucleotide reduction with activities that vary from 5% to 85% that of wt-β2. Each F[subscript n] Y·-β2 has been characterized by 9 and 130 GHz electron paramagnetic resonance and high-field electron nuclear double resonance spectroscopies. The hyperfine interactions associated with the 19 F nucleus provide unique signatures of each F[subscript n]Y· that are readily distinguishable from unlabeled Y·'s. The variability of the abiotic F[subscript n]Y pK a 's (6.4 to 7.8) and reduction potentials (-30 to +130 mV relative to Y at pH 7.5) provide probes of enzymatic reactions proposed to involve Y·'s in catalysis and to investigate the importance and identity of hopping Y·'s within redox active proteins proposed to protect them from uncoupled radical chemistry.National Institutes of Health (U.S.) (Grant GM29595)National Science Foundation (U.S.) (Grant 0645960

Prevalence of Yersinia enterocolitica across phases of swine production

The objective of this study was to describe the prevalence of Yersinia enterocolitica (YE) in different swine production phases. In this cross-sectional study, individual pigs on eight farrow-to-finish farms were sampled for YE by collection of both feces and oral-pharyngeal swab

Effect of cleaning and subtherapeutic chlortetracycline on Salmonella

The objective of this study was to evaluate the effect of stringent cleaning and subtherapeutic chlortetracycline (CTC) on Salmonella enterica (SE) prevalence in market age swine

Critical reflection of the advanced rehabilitation of an elite rugby league player sustaining a posterior Bankart lesion

The following is a critical description and discussion of the successful assessment and rehabilitation of a right shoulder posterior Bankart repair in an elite rugby league player. The rehabilitation follows accelerated, goal based guidelines, widely adopted in current sports practice but not well documented in the literature [Funk and Snow, 2007] and [Park et al., 2004]). The study serves to be the first critical discussion of such a regime