Characterization of a Tryptophan 6-Halogenase from Streptomyces albus and Its Regioselectivity Determinants

Tryptophan halogenases are found in diverse organisms and catalyze regiospecific halogenation. They play an important role in the biosynthesis of halogenated indole alkaloids, which are biologically active and of therapeutic importance. Here, a tryptophan 6-halogenase (SatH) from Streptomyces albus was characterized by using a whole-cell reaction system in Escherichia coli. SatH showed substrate specificity for chloride and bromide ions, leading to regiospecific halogenation at the C6-position of l -tryptophan. In addition, SatH exhibited higher performance in bromination than that of previously reported tryptophan halogenases in the whole-cell reaction system. Through structure-based protein mutagenesis, it has been revealed that two consecutive residues, A78/V79 in SatH and G77/I78 in PyrH, are key determinants in the regioselectivity difference between tryptophan 6- and 5-halogenases. Substituting the AV with GI residues switched the regioselectivity of SatH by moving the orientation of tryptophan. These data contribute to an understanding of the key residues that determine the regioselectivity of tryptophan halogenases.     

Enhancing cation exchange capacity of chars through ozonation

The use of ozone to increase the cation exchange capacity (CEC) of two chars produced from pyrolysis of Douglas fir (Pseudotsuga menziessii) and a control bituminous coal activated carbon (AC) is reported. Chars were produced from the wood fraction of Douglas fir (DFWC) and the bark (DFBC) at 500 °C using an auger driven reactor with a nitrogen sweep gas under mild vacuum. Five ozone treatment times, ranging from 5 min to 60 min, were investigated. The initial properties of each char were found to differ significantly from the other samples in terms of surface area, proximate composition, and elemental composition. DFWC did not show significant mass loss or temperature variation during ozone treatment; however, after 1 h of oxidation both DFBC and AC samples resulted in 20% and 30% mass loss, respectively, and reactor temperatures in excess of 60 °C. Analysis of the pore size distribution of each treatment shows that ozone treatment did not significantly affect small micropores after 30 min of treatment for any material, but did reduce the apparent surface area of mesopores. Increases in carboxylic groups were identified with ozone treatment and found to correlate strongly with changes in measured CEC. The formation of lactone was found to correlate positively with reactor temperature during oxidation. These results indicate that the properties of chars, including surface area, pore structure, and chemical composition, as well as reactor conditions strongly affect the ozone oxidation of chars.     

Comparative evaluation of the mesophilic and thermophilic biohydrogen production at optimized conditions using tequila vinasses as substrate

The objective of this work was to comparatively evaluate the production of biohydrogen (bio-H₂) from tequila vinasses at optimized mesophilic and thermophilic conditions and to elucidate the main metabolic routes involved. Optimal temperatures of 35 °C and 55 °C, and pH of 5.5 maximized the bio-H₂ production rates, 25.5 ± 0.01 NmL h⁻¹ and 169.9 ± 8.9 NmL h⁻¹ in the mesophilic and thermophilic regimens, respectively. During the operation of anaerobic sequencing batch reactors, the thermophilic process allowed a volumetric bio-H₂ production rate of 519 ± 13 NmL-H₂ L⁻¹ d⁻¹ equivalent to 750 ± 19 NmL-H₂ L_vinasse⁻¹, while the mesophilic one 448 ± 23 NmL-H₂ L⁻¹ d⁻¹ and 647 ± 33 NmL-H₂ L_vinasse⁻¹, respectively. Furthermore, the gas produced under thermophilic conditions showed high hydrogen content (86.5%). Finally, formate degradation and glucose fermentation to acetic and butyric acids were the main metabolic routes involved in bio-H₂ production under thermophilic conditions, while at mesophilic conditions, the lactate and formate degradation pathways governed.     

Significant population genetic structuring in Rhyzopertha dominica across Turkey: Biogeographic and practical implications

Rhyzopertha dominica is a key pest of stored grain. Understanding the movement of this beetle on broad geographic scales is crucial, particularly when developing strategies to prevent the spread of phosphine resistance. We assessed population genetic structuring in this pest across Turkey, using a combination of mitochondrial (cytochrome oxidase I) and microsatellite markers. In addition, we screened samples for Wolbachia, as this endosymbiont has previously been suggested to be associated with low mitochondrial genetic diversity in this beetle. Mitochondrial genetic diversity was low, with only six haplotypes identified. The genetic diversity was, however, substantially higher than that previously found in Australia or India, suggesting that R. dominica may have originated in the Middle East. Wolbachia were detected only at a single site, indicating they are not impacting the mitochondrial genetic diversity of R. dominica across Turkey. Microsatellite markers indicated there is significant geographic genetic structuring across Turkey, even among sites less than 100 km apart, suggesting there is little movement of beetles across regions within the country. This contrasts with the significantly higher levels of gene-flow found in Australia and the United States. We suggest that the limited movement of beetles across Turkey may be due to a combination of the historically localised agricultural practices (which limits anthropogenic movement among regions), and the mountainous landscape (which limits active flight among regions). Our results demonstrate that the movement of stored product pests may differ significantly across studies conducted in different countries. As a consequence, phosphine resistance management strategies must incorporate region specific information on the extent of beetle movement.     

Synchrotron X-ray microdiffraction analysis of proton irradiated polycrystalline diamond films

X-ray microdiffraction is a non-destructive technique that allows for depth-resolved, strain measurements with submicron spatial resolution. These capabilities make this technique promising for understanding the mechanical properties of MicroElectroMechancial Systems (MEMS). This investigation examined the local strain induced by irradiating a polycrystalline diamond thin film with a dose of 2 × 10¹⁷ H⁺/cm² protons. Preliminary results indicate that a measurable strain, on the order of 10^− 3, was introduced into the film near the End of Range (EOR) region of the protons.     

Solid–liquid phase equilibria of binary indole mixtures with some aromatic compounds using a solid–liquid–vapor equation-of-state

The separation of useful chemicals, such as indole from coal tar, is an important subject of research in the field of fossil energy, renewable energy, and utilization of byproducts and waste materials. High-pressure crystallization is one of the proposed separation methods. To understand the process requires detailed knowledge of solid–liquid phase behaviors. Solid–liquid phase equilibrium data of various binary mixtures of indole have been reported in the literature. In the present report, we analyze some of the experimental data with our unified solid–liquid–vapor equation-of-state to see whether our model can be useful for the calculation of solid–liquid equilibria in these organic systems. Good correlations of the data and predictions of phase behavior, at very high pressures, are demonstrated.     

Deformation behaviour of iron-doped alumina

Compressive creep tests in air were carried out on 1 cat.% Fe-doped alumina at a temperature T=1400 °C. Iron doping affected the plastic deformation by different ways in relation with Fe²⁺ cations population. Fe²⁺ cations sped up the deformation rates. FeAl₂O₄ spinel precipitates were identified and they were found (i) to interact with alumina grain boundaries (ii) to limit the grain growth within a range of strain. The Fe²⁺ cations underwent oxidation and this resulted in the dissolution of the some precipitates and in the decrease of deformation rates. It was suggested that deformation sped up this evolution through mass transport and that time was not a dominating parameter.     

An objective comparison of the cost effectiveness of three testing methods

Branch testing is a well established method for exercising software. JJ-path testing, whilst employed by some practitioners, is less popular, and the testing of JJ-pairs finds few adherents. In this paper an objective, practical study of the cost-effectiveness of these three testing methods is reported. The effectiveness of each method is assessed, in the presence of infeasible paths, not only on its ability to cover the specific structural element of code that it targets, but also on its ability to cover the structural elements targeted by the other two methods – the collateral coverage it achieves. The assessment is based on the results derived from experiments in which each of the three methods is applied to 35 units of program code.