Molecular mass calibration in size-exclusion chromatography of coal derivatives

A number of procedures have been investigated for the calibration of a size-exclusion chromatography (s.e.c.) column in the determination of molecular mass (MM) distributions of coal derivatives. The behaviour of narrow fractions of coal extracts in the MM range 200–3000 was compared with a variety of the more generally available calibration standards. Calibration with preparative s.e.c. subfractions of materials similar to those under study has been recommended. Polystyrene standards are satisfactory, however, for MM < 1000, but above this range other polymer standards should be sought. Universal and molar volume calibration do not apply to coal-derived materials.     

Comparison of the modelled thyroxine binding site in TBG with the experimentally determined site in transthyretin

The structure of cleaved thyroxine-binding globulin (TBG) hasbeen modelled on the crystal structure of cleaved 1-antitrypsin(a member of the serine proteinase inhibitor, serpin, superfamily)based on the high sequence homology exhibited by the two proteins.Particular attention was paid to the identification and modelledcharacteristics of the thyroxine binding site. The primary aimof the study was to compare the site qualitatively with thecrystallographically determined binding site of transthyretin,the other major transporter of thyroxine, in an attempt to explainthe higher binding affinity of the site compared with the knownthyrox ine binding site in transthyretin (10¹⁰ versus 10⁸ M^–1).The proposed binding site shares some similar characteristicswith the transthyretin binding site but also includes a clusterof aromatic residues which are entirely absent in transthyretin.It is proposed that this might account for the substantial differencein binding affinities.     

HDPE/LLDPE reactor blends with bimodal microstructures—Part II: rheological properties

Reactor blends of polyethylene/poly(ethylene-co-1-octene) resins with bimodal molecular weight and bimodal short chain branching distributions were synthesized in a two-step polymerization process. The compositions of these blends range from low molecular weight (LMW) homopolymer to high molecular weight (HMW) copolymer and vice versa HMW homopolymer to LMW copolymer. The shear flow characteristics of these polymers in the typical processing range mostly depend on the molecular weight and MWD of the polymer and are independent of the short chain branch content. From oscillatory shear measurements, it was observed that the viscosity of HMW polymers was reduced with the addition of LMW material. For the polymers produced with this two-step polymerization process, the LMW homopolymer and HMW copolymer blends and HMW homopolymer and LMW copolymer blends were melt miscible, despite the large viscosity differences of the pure components.     

Isolation and Characterisation of a Ferrirhodin Synthetase Gene from the Sugarcane Pathogen Fusarium sacchari

FSN1, a gene isolated from the sugar‐cane pathogen Fusarium sacchari, encodes a 4707‐residue nonribosomal peptide synthetase consisting of three complete adenylation, thiolation and condensation modules followed by two additional thiolation and condensation domain repeats. This structure is similar to that of ferricrocin synthetase, which makes a siderophore that is involved in intracellular iron storage in other filamentous fungi. Heterologous expression of FSN1 in Aspergillus oryzae resulted in the accumulation of a secreted metabolite that was identified as ferrirhodin. This siderophore was found to be present in both mycelium and culture filtrates of F. sacchari, whereas ferricrocin is found only in the mycelium, thus suggesting that ferricrocin is an intracellular storage siderophore in F. sacchari, whereas ferrirhodin is used for iron acquisition. To our knowledge, this is the first report to characterise a ferrirhodin synthetase gene functionally.     

Stepwise optimisation of enzyme production in solid state fermentation of waste bread pieces

When it is not consumed, bread presents a major source of food waste, both in terms of the amount and its economic value. However, bread also possesses the characteristics of an ideal substrate for solid state fermentation. Yet nearly all wasted bread ends up in landfill sites, where it is converted into methane by anaerobic digestion. Governments are finally taking action and, according to the EU Landfill Directive, for example, biodegradable municipal waste disposed into landfills must be decreased to 35% of 1995 levels, by 2020. Solid state fermentation of waste bread for the production of value added products is a novel idea, which could help with the achievement of this target. In this study, glucoamylase and protease production from waste bread pieces, via solid state fermentation, was investigated in detail. The optimum fermentation conditions for enzyme production were evaluated as, 20 mm particle size, 1.8 (w/w, db) initial moisture ratio, and duration of 144 h. Under these conditions, glucoamylase and protease activities reached up to 114.0 and 83.2 U/g bread (db), respectively. This study confirms that waste bread could be successfully utilised as a primary raw material in cereal based biorefineries.     

Exchanging the substrate specificities of pyruvate decarboxylase from Zymomonas mobilis and benzoylformate decarboxylase from Pseudomonas putida

Pyruvate decarboxylase from Zymomonas mobilis (PDC) and benzoylformate decarboxylase from Pseudomonas putida (BFD) are thiamine diphosphate-dependent enzymes that decarboxylate 2-keto acids. Although they share a common homotetrameric structure they have relatively low sequence similarity and different substrate spectra. PDC prefers short aliphatic substrates whereas BFD favours aromatic 2-keto acids. These preferences are also reflected in their carboligation reactions. PDC catalyses the conversion of benzaldehyde and acetaldehyde to (R)-phenylacetylcarbinol and predominantly (S)-acetoin, whereas (R)-benzoin and mainly (S)-2-hydroxypropiophenone are the products of BFD catalysis. Comparison of the X-ray structures of both enzymes identified two residues in each that were likely to be involved in determining substrate specificity. Site-directed mutagenesis was used to interchange these residues in both BFD and PDC. The substrate range and kinetic parameters for the decarboxylation reaction were studied for each variant. The most successful variants, PDCI472A and BFDA460I, catalysed the decarboxylation of benzoylformate and pyruvate, respectively, although both variants now preferred the long-chain aliphatic substrates, 2-ketopentanoic and 2-ketohexanoic acid. With respect to the carboligase activity, PDCI472A proved to be a real chimera between PDC and BFD whereas BFDA460I/F464I provided the most interesting result with an almost complete reversal of the stereochemistry of its 2-hydroxypropiophenone product.     

Obesity: Prevalence,Theories, Medical Consequences,Management, and Research Directions

Obesity and its associated disorders are a growing epidemic across the world. Many genetic, physiological, and behavioral factors play a role in the etiology of obesity. Diet and exercise are known to play a valuable role in the treatment and prevention of obesity and associated disorders such as hypertension, heart disease, and diabetes. Therefore, the purpose of this review is to examine the prevalence, etiology, consequences, and treatment of obesity.     

Electro-Oxidation of Carbonate in Aqueous Solution on a Platinum Rotating Ring Disk Electrode

The kinetics of electro-oxidation of carbonate aqueous solution on a platinum electrode were examined by a rotating ring (platinum) – disk (platinum) electrode in aqueous sodium carbonate/bicarbonate solutions (25 °C, pH = 10.75) with the peroxide stabilizing additives sodium silicate, magnesium sulphate and sodium DTPA such as are used in brightening mechanical wood pulp. A theoretical model based on the experimental data is proposed to describe this electro-oxidation process. The model shows consistency between the experimental results and the expectation from the proposed reaction sequence. The first-order rate constants at 25 °C, pH = 10.75 in 1.13 M (Na₂CO₃ + NaHCO₃) for carbonate electro-oxidation to percarbonate (a.k.a. peroxidicarbonate C₂O₆^2-), percarbonate hydrolysis to hydrogen peroxide and hydrogen peroxide electro-oxidation to oxygen are estimated from the experimental data, respectively, as (1.43±0.1) × 10⁻¹¹cm s⁻¹ (extrapolated to the equilibrium electrode potential which is estimated as ca. 0.21 (vs SCE)), (1.1 ± 0.2) × 10⁻² s⁻¹ and (5.6±0.5) × 10⁻⁸cm s⁻¹ (extrapolated to the equilibrium electrode potential of −0.019 V (vs SCE)).     

Responses of the Crab <Emphasis Type="Italic">Heterozius rotundifrons</Emphasis>to Heterospecific Chemical Alarm Cues: Phylogeny vs. Ecological Overlap

The big-handed brachyuran crab Heterozius rotundifronsextends the time spent in its anti-predator posture, limb extended posture, if exposed to chemical cues from crushed conspecifics. In this study, we tested whether crabs also respond to chemical cues from crushed heterospecific crabs, and if so, whether phylogenetic relations or ecological overlap is more important in influencing the duration of the anti-predator posture. Chemical cues from two other brachyuran crabs (Cyclograpsus lavauxi and Hemigrapsus sexdentatus), which do not overlap directly in ecological distribution with H. rotundifrons, elicited a duration of the anti-predator posture that was indistinguishable from that produced by conspecific chemical cues. In contrast, chemical cues from two anomuran crabs (Petrolisthes elongatus and Pagurus novizealandiae) that overlap in ecological distribution with H. rotundifrons, elicited durations of the antipredator posture that were significantly shorter than those of either conspecifics or more closely related crab species. Thus, phylogenetic relationship seems to be more important than ecological overlap in influencing anti-predator behavior in H. rotundifrons.