Search results for: Wayde Veldman

Authors: Wayde Veldman, Ozlem T. Bishop, Igor Polikarpov

Abstract:

In bacteria, mono and disaccharides are phosphorylated during uptake into the cell via the widely used phosphoenolpyruvate (PEP)-dependent phosphotransferase transport system. As an initial step in the phosphorylated disaccharide metabolism pathway, certain glycoside hydrolase family 1 (GH1) enzymes play a crucial role in releasing phosphorylated and non-phosphorylated monosaccharides. However, structural determinants for the specificity of these enzymes still need to be clarified. GH1 enzymes are known to have a wide array of functions. According to the CAZy database, there are twenty-one different enzymatic activities in the GH1 family. Here, the structure and substrate specificity of a GH1 enzyme from Bacillus licheniformis, hereafter known as BlBglH, was investigated. The sequence of the enzyme BlBglH was compared to the sequences of other characterized GH1 enzymes using sequence alignment, sequence identity calculations, phylogenetic analysis, and motif discovery. Through these various analyses, BlBglH was found to have sequence features characteristic of the 6-phospho-β-glucosidase activity enzymes. Additionally, motif and structure comparisons of the three most commonly studied GH1 enzyme-activities revealed a shared loop amongst the different structures that consist of different sequence motifs – this loop is thought to guide specific substrates (depending on activity) towards the active-site. To further affirm BlBglH enzyme activity, molecular docking and molecular dynamics simulations were performed. Docking was carried out using 6-phospho-β-glucosidase enzyme-activity positive (p-Nitrophenyl-beta-D-glucoside-6-phosphate) and negative (p-Nitrophenyl-beta-D-galactoside-6-phosphate) control ligands, followed by 400 ns molecular dynamics simulations. The positive-control ligand maintained favourable interactions within the active site until the end of the simulation. The negative-control ligand was observed exiting the enzyme at 287 ns. Binding free energy calculations showed that the positive-control complex had a substantially more favourable binding energy compared to the negative-control complex. Jointly, the findings of this study suggest that the BlBglH enzyme possesses 6-phospho-β-glucosidase enzymatic activity.

Keywords: 6-P-β-glucosidase, glycoside hydrolase 1, molecular dynamics, sequence analysis, substrate specificity

Procedia PDF Downloads 93

Authors: A van Onselen, C. M. Walsh, F. J. Veldman, C. Brand

Abstract:

Objectives: To determine the nutritional status and risk factors associated with women practicing geophagia in QwaQwa, South Africa. Materials and Methods: An observational epidemiological study design was adopted which included an exposed (geophagia) and non-exposed (control) group. A food frequency questionnaire, anthropometric measurements and blood sampling were applied to determine nutritional status of participants. Logistic regression analysis was performed in order to identify factors that were likely to be associated with the practice of geophagia. Results: The mean total energy intake for the geophagia group (G) and control group(C) were 10324.31 ± 2755.00 kJ and 10763.94 ± 2556.30 kJ respectively. Both groups fell within the overweight category according to the mean body mass index (BMI) of each group (G= 25.59 kg/m2; C= 25.14 kg/m2). The mean serum iron levels of the geophagia group (6.929 μmol/l) were significantly lower than that of the control group (13.75 μmol/l) (p = 0.000). Serum transferrin (G=3.23g/l; C=2.7054g/l) and serum transferrin saturation (G=8.05%; C=18.74%) levels also differed significantly between groups (p=0.00). Factors that were associated with the practice of geophagia included haemoglobin (Odds ratio (OR):14.50), serum-iron (OR: 9.80), serum-ferritin (OR: 3.75), serum-transferrin (OR: 6.92) and transferrin saturation (OR: 14.50). A significant negative association (p=0.014) was found between women who were wage-earners and those who were not wage-earners and the practice of geophagia (OR: 0.143; CI: 0.027; 0.755). These findings seem to indicate that a permanent income may decrease the likelihood of practising geophagia. Key findings: Geophagia was confirmed to be a risk factor for iron deficiency in this community. The significantly strong association between geophagia and iron deficiency emphasizes the importance of identifying the practice of geophagia in women, especially during their child bearing years. Further research to establish whether the practice of geophagia is a cause of iron-deficiency, or whether it is the consequence thereof, would give a clearer view on how to recognise and treat the condition.

Keywords: geophagia, iron deficiency anaemia, dietary intake, anthropometry

Procedia PDF Downloads 316