Using photoelectron elliptical dichroism (PEELD) to determine real-time variation of enantiomeric excess

In this work, the photoionization of chiral molecules by an elliptically polarized, high repetition rate, femtosecond laser is probed. The resulting 3D photoelectron angular distribution shows a strong forward–backward asymmetry, which is highly dependent not only on the molecular structure but also on the ellipticity of the laser pulse. By continuously varying the laser ellipticity, we can observe molecular and enantiomer changes in real time at a previously unseen speed and precision. The technique allows enantiomeric excess of a pure compound to be measured with a 5% precision within 3 s, and a 10-min acquisition yields a precision of 0.4%. The isomers camphor and fenchone can be easily distinguished, unlike with conventional mass spectrometry. Preliminary results for the pharmaceutically interesting ibuprofen are also given, showing the capability of photoionization as a means of distinguishing larger molecular systems.     

Molecular cloning and expression of a Bacillus subtilis β-glucanase gene in Escherichia coli

A Bacillus subtilis gene coding for an endo-β-1,3-1,4-glucanase has been transferred to Escherichia coli by molecular cloning using bacteriophage λ and plasmid vectors. The gene is contained within a 1.6-kb EcoRI-PvuI DNA fragment and directs the synthesis in E. coli of a β-glucanase which specifically degrades barley glucan and lichenan. A novel dye-staining method has been developed to detect β-glucanase activity in colonies on agar plates.     

Quantifying spatiotemporal changes in a sagebrush ecosystem in relation to energy development

Energy development has been occurring in the intermountain western United States for over a century, yet few studies have attempted to spatially quantify the impacts of this disturbance on native ecosystems. We used temporal remotely sensed data for the Pinedale Anticline Project Area (PAPA) in western Wyoming, a region that has experienced increased natural gas development within the past 10 yr, to quantify the spatiotemporal distribution of Wyoming big sagebrush Artemisia tridentata, natural gas development, and other landcover types. Our analyses included 5 Landsat Thematic Mapper (TM) images of the PAPA over a 22‐yr period (1985–2006). We determined whether Wyoming big sagebrush spatiotemporal patterns were associated with natural gas development or other landcover types. We also developed a footprint model to determine the direct and indirect impacts of natural gas development on the distribution of Wyoming big sagebrush habitats. Over the 22‐yr period, we observed an inverse relationship between the amount of Wyoming big sagebrush habitat and natural gas development. During this time, Wyoming big sagebrush habitat declined linearly at a rate of 0.2% yr^?1 (4.5% total net loss), whereas natural gas development increased exponentially at a rate of 20% yr^?1 (4800% total net increase). Our evaluation indicated that, by 2006, natural gas development directly impacted 2.7% (1750 ha) of original Wyoming big sagebrush habitat. Indirect impacts, quantified to account for degraded habitat quality, affected as much as 58.5% (assuming 1000‐m buffers) of the original Wyoming big sagebrush habitat. Integrating assessments of the direct and indirect impacts will yield a better elucidation of the overall effects of disturbances on ecosystem function and quality.     

Isolation of deacetoxycephalosporin C from fermentation broths of Penicillium chrysogenum transformants: construction of a new fungal biosynthetic pathway.

Deacetoxycephalosporin C (DAOC), a precursor of cephalosporins excreted by Cephalosporium and Streptomyces species, has been produced in Penicillium chrysogenum transformed with DNA containing a hybrid penicillin N expandase gene (cefEh) and a hybrid isopenicillin N epimerase gene (cefDh). DAOC from a P. chrysogenum transformant was identified by ultraviolet light (UV), high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) and mass spectrum analyses. P. chrysogenum transformed with DNA containing cefEh without cefDh did not produce DAOC. Untransformed P. chrysogenum produced penicillin V (phenoxymethylpenicillin) but not DAOC. Transformants also produced penicillin V but, in general, less than untransformed P. chrysogenum. The cefEh and cefDh genes were constructed by replacing the open reading frame (ORF) of cloned P. chrysogenum pcbC and penDE genes with the ORF of the Streptomyces clavuligerus expandase gene, cefE, and the ORF of the Streptomyces lipmanii epimerase gene, cefD, respectively. Analyses of representative transformants suggested that production of DAOC occurred via cefEh and cefDh genes stably integrated in the P. chrysogenum genome. DNA from untransformed P. chrysogenum did not hybridize to cefE or cefD gene probes.     

Microbial growth on hydrocarbons: terminal branching inhibits biodegradation

A variety of octane-utilizing bacteria and fungi were screened for growth on some terminally branched dimethyloctane derivatives to explore the effects of iso- and anteiso-termini on the biodegradability of such hydrocarbons. Of 27 microbial strains tested, only 9 were found to use any of the branched hydrocarbons tested as a sole carbon source, and then only those hydrocarbons containing at least one iso-terminus were susceptible to degradation. Anteiso-or isopropenyl termini prevented biodegradation. None of the hydrocarbonoclastic yeasts tested was able to utilize branched-hydrocarbon growth sustrates. In the case of pseudomonads containing the OCT plasmid, whole-cell oxidation of n-octane was poorly induced by terminally branched dimethyloctanes. In the presence of a gratuitous inducer of the octane-oxidizing enzymes, the iso-branched 2,7-dimethyloctane was slowly oxidized by whole cells, whereas the anteiso-branched 3,6-dimethyloctane was not oxidized at all. This microbial sampling dramatically illustrated the deleterious effect of alkyl branching, especially anteiso-terminal branching, on the biodegradation of hydrocarbons.