Extraction of hemoglobin with calixarenes and biocatalysis in organic media of the complex with pseudoactivity of peroxidase

The present work involves the use of p-tert-butylcalix[4,6,8]arene carboxylic acid derivatives ((t)Butyl[4,6,8]CH2COOH) for selective extraction of hemoglobin. All three calixarenes extracted hemoglobin into the organic phase, exhibiting extraction parameters higher than 0.90. Evaluation of the solvent accessible positively charged amino acid side chains of hemoglobin (PDB entry 1XZ2) revealed that there are 8 arginine, 44 lysine and 30 histidine residues on the protein surface which may be involved in the interactions with the calixarene molecules. The hemoglobin-(t)Butyl[6]CH2COOH complex had pseudoperoxidase activity which catalysed the oxidation of syringaldazine in the presence of hydrogen peroxide in organic medium containing chloroform. The effect of pH, protein and substrate concentrations on biocatalysis was investigated using the hemoglobin-(t)Butyl[6]CH2COOH complex. This complex exhibited the highest specific activity of 9.92 x 10(-2) U mg protein(-1) at an initial pH of 7.5 in organic medium. Apparent kinetic parameters (V'(max), K'(m), k'(cat) and k'(cat)/K'(m)) for the pseudoperoxidase activity were determined in organic media for different pH values from a Michaelis-Menten plot. Furthermore, the stability of the protein-calixarene complex was investigated for different initial pH values and half-life (t(1/2)) values were obtained in the range of 1.96 and 2.64 days. Hemoglobin-calixarene complex present in organic medium was recovered in fresh aqueous solutions at alkaline pH, with a recovery of pseudoperoxidase activity of over 100%. These results strongly suggest that the use of calixarene derivatives is an alternative technique for protein extraction and solubilisation in organic media for biocatalysis.

Py-GC/MS(FID) assessed behavior of polysaccharides during kraft delignification of Eucalyptus globulus heartwood and sapwood

Eucalyptus globulus heartwood, sapwood and their delignified samples by kraft pulping at 130, 150 and 170 degrees C along time were characterized in respect to total carbohydrates by Py-GC/MS(FID). No significant differences between heartwood and sapwood were found in relation to pyrolysis products and composition. The main wood carbohydrate derived pyrolysis compounds were levoglucosan (25.1%), hydroxyacetaldehyde (12.5%), 2-oxo-propanal (10.3%) and acetic acid (8.7%). Levoglucosan decreased during the early stages of delignification and increased during the bulk and residual phases. Acetic acid decreased hydroxyacetaldehyde and 2-oxo-propanal increased, and 2-furaldehyde and hydroxypropanone remained almost constant during delignification. The C/L ratio was 3.2 in wood and remained rather constant in the first pulping periods until a loss of 15-25% in carbohydrate and 60% in lignin. Afterwards it increased sharply until 44 that correspond to the removal of 25-35% of carbohydrates and 95% of lignin. The pulping reactive selectivity to lignin vs. polysaccharides was the same for sapwood and heartwood. (C) 2013 Elsevier B.V. All rights reserved.

Exposure to fungi in cork: potential occupational hazard

Cork is a light, porous and impermeable material extracted from the bark of some trees. It is in manufacture of stoppers for wine bottles the main application of cork. It is estimated that the area occupied by cork oaks in the Iberian Peninsula is around 33% in Portugal and 23% in Spain. The world production of cork is focused in the south Europe, with Portugal being the most important producer followed by Spain. According to Companies Directory more than 100 manufactories from Portugal has their branch associated with the preparation and fabrication of cork. Cork workers are at risk for developing diseases of the respiratory tract such as occupational asthma and Suberosis, a form of pulmonary hypersensitivity due to repeated exposure to mouldy cork dust. In this review study papers from 2000 were analyzed to better understand which fungi species are associated with occupational disease in cork workers. The most prevalent fungi species in these workers that are associated with those occupational diseases are Penicilliumglabrum, Chrysoniliasitophila and Trichodermalongibrachiatum. Therefore, a specific knowledge about occupational exposure to fungi in the cork industry is the key to better understand the related diseases and to define preventive measures. Given the importance of this occupational setting in Portugal is essential to evaluate the combined exposure of fungi and particles and their metabolites. Further studies concerning exposure assessment to fungi and particles in the cork industry must be developed.