Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis through Reduced ATPase Activity

Translation elongation is mediated by ribosomes and multiple soluble factors, many of which are conserved across bacteria and eukaryotes. During elongation, eukaryotic elongation factor 1A (eEF1A; EF-Tu in bacteria) delivers aminoacylated-tRNA to the A-site of the ribosome, whereas eEF2 (EF-G in bacteria) translocates the ribosome along the mRNA. Fungal translation elongation is striking in its absolute requirement for a third factor, the ATPase eEF3. eEF3 binds close to the E-site of the ribosome and has been proposed to facilitate the removal of deacylated tRNA from the E-site. eEF3 has two ATP binding cassette (ABC) domains, the second of which carries a unique chromodomain-like insertion hypothesized to play a significant role in its binding to the ribosome. This model was tested in the current study using a mutational analysis of the Sac7d region of the chromodomain-like insertion. Specific mutations in this domain result in reduced growth rate as well as slower translation elongation. In vitro analysis demonstrates that these mutations do not affect the ability of eEF3 to interact with the ribosome. Kinetic analysis revealed a larger turnover number for ribosomes in comparison to eEF3, indicating that the partial reactions involving the ribosome are significantly faster than that of eEF3. Mutations in the chromodomain-like insertion severely compromise the ribosome stimulated ATPase of eEF3, strongly suggesting that it exerts an allosteric effect on the hydrolytic activity of eEF3. The chromodomain-like insertion is, therefore, vital to eEF3 function and may be targeted for developing novel antifungal drugs.     

Chaos game representation for comparison of whole genomes

Background  

Chaos game representation of genome sequences has been used for visual representation of genome sequence patterns as well as alignment-free comparisons of sequences based on oligonucleotide frequencies. However the potential of this representation for making alignment-based comparisons of whole genome sequences has not been exploited.     

Design and synthesis of tricyclic sulfones as γ-secretase inhibitors with greatly reduced Notch toxicity

A novel series of tricyclic γ-secretase inhibitors was designed and synthesized via a conformational analysis of literature compounds. The preliminary results have shown that compounds in this new series have much improved in vitro potency and in vivo profiles. More importantly, they have greatly reduced Notch related toxicity that was associated with previous γ-secretase inhibitors.     

Torrefaction: a sustainable method for transforming of agri-wastes to high energy density solids (biocoal)

Reviews in Environmental Science and Bio/Technology - The depletion of fossil fuel reserves and greenhouse gas emissions led to limit the use of fossil fuels, including natural gas, coal, or...     

S-adenosyl-L-methionine activates actinorhodin biosynthesis by increasing autophosphorylation of the Ser/Thr protein kinase AfsK in Streptomyces coelicolor A3(2)

S-Adenosyl-L-methionine (SAM) is one of the major methyl donors in all living organisms. The exogenous treatment with SAM leads to increased actinorhodin production in Streptomyces coelicolor A3(2). In this study, mutants from different stages of the AfsK-AfsR signal transduction cascade were used to test the possible target of SAM. SAM had no significant effect on actinorhodin production in afsK, afsR, afsS, or actII-open reading frame 4 (ORF4) mutant. This confirms that afsK plays a critical role in delivering the signal generated by exogenous SAM. The afsK-pHJL-KN mutant did not respond to SAM, suggesting the involvement of the C-terminal of AfsK in binding with SAM. SAM increased the in vitro autophosphorylation of kinase AfsK in a dose-dependent manner, and also abolished the effect of decreased actinorhodin production by a Ser/Thr kinase inhibitor, K252a. In sum, our results suggest that SAM activates actinorhodin biosynthesis in S. coelicolor M130 by increasing the phosphorylation of protein kinase AfsK.     

ZnWO4:Eu3+ phosphor with intense blue LED excitation: photoluminescence and electron density distribution analysis

A high intensity 464 nm excitable ZnWO₄:Eu³⁺ red‐emitting phosphor for warm white lighting applications was prepared using a solid‐state reaction method by varying the dopant Eu³⁺ concentration. Crystalline purity and phase identification was confirmed and revealed using powder X‐ray diffraction and Rietveld refinement analysis. The surface morphology of Zn_1‐xEu_xWO₄ (x = 0, 0.01, 0.02, 0.03, 0.04 and 0.05) was examined using scanning electron microscopy (SEM) techniques. From SEM analysis, the ZnWO₄:Eu³⁺ phosphor prepared at 1–3% molar Eu³⁺ concentrations exhibited a small pebble‐like morphology with a smooth surface. On increasing the molar concentration of Eu³⁺ to >3%, the pebble stone morphology disappeared and a large, smooth irregular polygon‐shaped granular‐like morphology was obtained. Of the higher mol% Eu³⁺, the 4% Eu³⁺‐doped ZnWO₄ showed the best photoluminescence properties with high intensity and sharp excitation at 395 and 464 nm, followed by red emission centred at 615 nm with excellent CIE coordinates (x = 0.58 and y = 0.41) in the core red region. Elemental composition and chemical state analysis were carried out for the 4% Eu³⁺‐doped ZnWO₄ phosphor using X‐ray photoelectron spectroscopy and energy dispersive X‐ray spectroscopy studies. Based on all the above analyses, the Eu³⁺‐doped ZnWO₄ phosphor was found to be a very efficient red‐emitting phosphor under near‐UV light as well as under visible light excitation and could be used for white LED and field emissive displays applications.     

Epigallocatechin gallate incorporation into lignin enhances the alkaline delignification and enzymatic saccharification of cell walls

ABSTRACT: BACKGROUND: Lignin is an integral component of the plant cell wall matrix but impedes the conversion of biomass into biofuels. The plasticity of lignin biosynthesis should permit the inclusion of new compatible phenolic monomers such as flavonoids into cell wall lignins that are consequently less recalcitrant to biomass processing. In the present study, epigallocatechin gallate (EGCG) was evaluated as a potential lignin bioengineering target for rendering biomass more amenable to processing for biofuel production. RESULTS: In vitro peroxidase-catalyzed polymerization experiments revealed that both gallate and pyrogallyl (B-ring) moieties in EGCG underwent radical cross-coupling with monolignols mainly by beta--O--4-type cross-coupling, producing benzodioxane units following rearomatization reactions. Biomimetic lignification of maize cell walls with a 3:1 molar ratio of monolignols and EGCG permitted extensive alkaline delignification of cell walls (72 to 92 %) that far exceeded that for lignified controls (44 to 62 %). Alkali-insoluble residues from EGCG-lignified walls yielded up to 34 % more glucose and total sugars following enzymatic saccharification than lignified controls. CONCLUSIONS: It was found that EGCG readily copolymerized with monolignols to become integrally cross-coupled into cell wall lignins, where it greatly enhanced alkaline delignification and subsequent enzymatic saccharification. Improved delignification may be attributed to internal trapping of quinone-methide intermediates to prevent benzyl ether cross-linking of lignin to structural polysaccharides during lignification, and to the cleavage of ester intra-unit linkages within EGCG during pretreatment. Overall, our results suggest that apoplastic deposition of EGCG for incorporation into lignin would be a promising plant genetic engineering target for improving the delignification and saccharification of biomass crops.     

Adaptive and cross resistance to cadmium (II) and zinc (II) by Pseudomonas aeruginosa BC15

Cadmium and zinc appear in the combined forms and they are co-pollutants. Cd is the most hazardous metal ion for human beings and causes renal dysfunction, liver and lungs damage, bone degeneration and blood damage. Though Zn is an essential nutrient, excess of Zn is toxic. Biological process was more important because conventional methods fail to remediate these pollutants due to high costs and less affordability. The screening and understanding of the functioning of microorganism plays an important role in removal and recovery of metals from heavy-metal-polluted water and soil. In our study, the strain Pseudomonas aeruginosa BC15 was isolated from oil-mill-treated waste water and it showed to be highly resistant to 6 mM Cd and 20 mM Zn in the solid and liquid media. The growth studies of BC15 strain in the medium without induction exhibited high tolerable capacity when compared to other microbes. Pretreatment of P. aeruginosa BC15 with sub-lethal concentrations of Cd induced adaptive resistance to lethal doses of Cd. Cadmium-induced cells also showed cross resistance to lethal concentration of zinc. The organism had high resistance against Cd and Zn. This has been clearly proven through biosorption studies: Cd was absorbed up to 62% and Zn about 60% in single solution, whereas in binary solution Cd was biosorbed up to 82% and Zn 85%. In conclusion, this study reveals the significance of using the strain P. aeruginosa BC15 in the bioremediation of Cd and Zn from industrial waste water and contaminated soil.